Dissertations / Theses on the topic 'Cantharidae'

A tribo Chauliognathini, na atual definicão, é composta por ca. 650 espécies em 14 gêneros: Chauliognathus, Daiphron, Microdaiphron, Malthesis, Psilorhynchus, Belotus, Maronius, Paramaronius, Maroniodes, Lobetus, Pseudolobetus, Macromalthinus, Malthoichthyurus e Malthinocantharis - estes dois últimos considerados incertae sedis. Caracteriza-se, principalmente, pela forte assimetria do abdômen e genitália dos machos, presença de sutura epistomal e ausência de esporões tibiais. Tradicionalmente, a classificação da subfamília Chauliognathinae e suas duas tribos era baseada principalmente no comprimento dos élitros: os gêneros de élitros longos eram agrupados em Chauliognathini, enquanto os de élitros curtos eram posicionados em Ichthyurini. Na atual classificação os gêneros foram redistribuídos e as tribos definidas com base na morfologia dos segmentos genitais e genitálias dos machos. Para testar a monofilia de Chauliognathini e verificar as relações entre seus gêneros foi feita uma análise cladística da tribo com 138 caracteres morfológicos envolvendo 45 espécies representantes de todos os gêneros de Chauliognathini e alguns de Ichthyurini, Silinae e Cantharinae. Foram encontradas 48 árvores igualmente mais parcimoniosas, com 341 passos de transformação. Demonstrou-se que Chauliognathini não é monofilética. Dois clados principais foram encontrados, agrupando os gêneros de acordo com a classificação tradicional de Chauliognathinae. O clado que representa a tribo Ichthyurini surge dentre os gêneros de Chauliognathini de élitros curtos. Os gêneros Chauliognathus, Daiphron, Malthesis, Belotus e Lobetus também são parafiléticos. Malthopterus pertence ao grupo formado pelos gêneros de élitros longos, enquanto Malthinocantharis deve ser excluído de Chauliognathini e posicionado em Silinae
The tribe Chauliognathini, in its current definition, is composed of approximately 650 species in 14 genera: Chauliognathus, Daiphron, Microdaiphron, Malthesis, Psilorhynchus, Belotus, Maronius, Paramaronius, Maroniodes, Lobetus, Pseudolobetus, Macromalthinus, Malthoichthyurus e Malthinocantharis - the two latter as incertae sedis. They are characterized by the strong asimmetry of the abdomen and male genitalia, presence of an epistomal suture and absence of tibial spurs. Traditionally, the classification of the subfamily Chauliognathinae and its two tribes were based mainly on elytral length: long-elytra genera were grouped in Chauliognathini, while short-elytra genera were classified in Ichthyurini. On present classification, the genera were redistributed and the tribes were defined based on the morphology of male genital segments and genitalia. To test the monophyly of Chauliognathini and to verify the relationships among its genera, it was conducted a cladistic analysis of the tribe with 138 morphological characters involving 45 species representative of all the genera of Chauliognathini, and some of Ichthyurini, Silinae and Cantharinae. The 48 equally most parsimonious trees (length=341) showed that Chauliognathini is not monophyletic. Two main clades were found, grouping the genera according to the traditional classification of Chauliognathinae. The clade that represents the tribe Ichthyurini is retrieved in a clade composed by the short-elytra genera of Chauliognathini. The genera Chauliognathus, Daiphron, Malthesis, Belotus and Lobetus are also paraphyletic. Malthopterus belongs to the group composed by long-elytra genera, while Malthinocantharis must be excluded from Chauliognathini and positioned in Silinae

A subfamília Chauliognathinae Champion, 1914 é dividida em duas tribos: Chauliognathini LeConte, 1861 e Ichthyurini Champion, 1915. Na classificação de Miskimen (1961) Chauliognathini é formada por espécies que possuem élitros longos, recobrindo totalmente ou a maior parte do abdômen, enquanto nas espécies de Ichthyurini os élitros são muito curtos, expondo as asas e os tergitos abdominais. Em uma classificação alternativa, Magis & Wittmer (1974) propõem que as tribos sejam definidas principalmente com base em caracteres dos abdomens e genitálias dos machos. Nessa proposta, parte dos gêneros de Ichthyurini foi transferida para Chauliognathini. No entanto, uma análise cladística com o objetivo de testar a monofilia de Chauliognathini sensu Magis & Wittmer indicou que este não é monofilético, recuperando os grupos conforme a proposta de Miskimen (Biffi 2012). A classificação dos gêneros de Ichthyurini está relativamente bem resolvida, com quase todos os gêneros revisados. Por outro lado, a taxonomia de Chauliognathini é caótica, com poucos gêneros que não representam suficientemente a grande diversidade morfológica do grupo. As cerca de 900 espécies e subespécies estão alocadas principalmente em um único gênero, Chauliognathus Hentz, 1830, cuja definição pouco precisa abrange todas as espécies de Chauliognathini. É apresentada uma nova proposta de homologias para os caracteres do edeago da subfamília Chauliognathinae, principal estrutura utilizada por alguns autores para delimitar as tribos e gêneros. Em seguida, é proposta uma análise cladística da tribo Chauliognathini com o objetivo de elucidar o posicionamento filogenético de suas espécies e apresentar caracteres que possam sustentar uma nova classificação dos gêneros. Foram amostradas espécies representantes da grande diversidade morfológica e da ampla distribuição geográfica, incluindo as espécies-tipo de todos os gêneros e subgêneros. Os resultados confirmam a monofilia de Chauliognathini e Ichthyurini como grupos-irmãos, conforme a proposta de Miskimen (1961). São discutidas as possibilidades de redefinição ou proposição de novos gêneros, com a redistribuição das espécies. Por fim, como um primeiro passo nos futuros trabalhos de revisões dos gêneros da tribo, é apresentada uma revisão taxonômica de Microdaiphron Pic, 1926. Dez espécies são reconhecidas como válidas, sendo duas novas, e 23 espécies ou subespécies são propostos como sinônimos. São apresentadas redescrições, ilustrações e um mapa de distribuição para todas as espécies
The subfamily Chauliognathini Champion, 1914 is composed of two tribes: Chauliognathini LeConte, 1861 e Ichthyurini Champion, 1915. In Miskimen\'s (1961) classification the species of each tribe differ in terms of the length of elytra, which may are long, covering the abdomen (Chauliognathini) or very short, exposing the wings and abdominal tergites (Ichthyurini). Alternatively, Magis & Wittmer (1974) proposed that the tribes should be defined based on characters of the abdomen and genitalia of males. In this proposal, part of the Ichthyurini genera was transferred to Chauliognathini. However, a cladistic analysis conducted in order to test the monophyly of Chauliognathini sensu Magis & Wittmer indicated that the group is not monophyletic, recovering the groups according to Miskimen\'s proposal (Biffi 2012). The classification of the genera in Ichthyurini is reliable, with taxonomic revisions available for most of them. However, the taxonomy of Chauliognathini is chaotic, with few genera that do not represent the group\'s morphological diversity accordingly. The approximately 900 species and subspecies are mainly allocated in a single genus, Chauliognathus Hentz, 1830, whose imprecise definition covers all the species of Chauliognathini. A new proposal of homologies for the aedeagus of Chauliognathinae is presented. This is the main structure used by some authors to delimit the tribes and genera. Then, a cladistic analysis of Chauliognathini is proposed in order to verify the phylogenetic positioning of its species and to present morphological characters that may support new classifications for the genera. Species representative of the great morphological diversity and the wide geographic distribution were sampled, including the type species of all genera and subgenera. The results confirm the monophyly of Chauliognathini and Ichthyurini as sister groups, according to Miskimen\'s proposal. We discuss the possibilities of redefining or proposing new genera, with a redistribution of species. Finally, a taxonomic revision of Microdaiphron Pic, 1926, is presented. Ten species are recognized as valid, two of them proposed as new species, and 23 species or subspecies are proposed as synonyms. Redescriptions, illustrations and a distribution map are presented for all species

Background: Despite increasing incidence and morbidity globally, hepatocellular carcinoma (HCC) remains a big challenge clinically. The difficulty to treat HCC is largely due to non-specific chemotherapy causing life-threatening toxicity and severe drug-related adverse effects. Extensive studies on targeted drug delivery systems (DDS) have revealed a great potential in specific delivery of chemotherapeutics for cancer treatment, which should be a way to overcome the limitations of conventional chemotherapy. Cantharidin (CTD) is a natural product from Chinese medicine showing a great potency but narrow therapeutic window with high toxicity. Its therapeutic potential is proposed to be improved with nanoliposomal encapsulation. To explore the potential of this liposomal delivery system for HCC treatment, in this study we developed and characterized liposomal carriers with CTD encapsulated and liposomal surface modified for targeted delivery to the HCC models in vitro and in vivo. Methods: In the present study, liposomal delivery system was developed with cantharidin (CTD) encapsulated as anticancer assembly for HCC treatment. Firstly, in order to demonstrate the feasibility of liposomal encapsulation for CTD, the plain liposomal CTD was prepared and the anticancer effects were evaluated in vitro and in vivo with comparison to the free CTD formulation (Chapter 2). Then, to achieve specific penetrability of the liposomal CTD for HCC, it was further modified with a cancer cell specific penetrating peptide BR2, and its superior penetrability was evaluated on both in vitro monolayer and 3D HepG2 cells including MTT assay, cellular uptake, internalization, tumor spheroid penetration and inhibition, and in vivo subcutaneous HCC mice model (Chapter 3). Finally, the dual-functionalized liposomes with BR2 and anti-carbonic anhydrase IX (CA IX) antibody were achieved for more efficient delivery with specific penetrating and targeting properties on orthotopic HCC model (Chapter 4). Results: The key results of the study are: (1) liposomal CTD can augment the anti-proliferative effects of CTD, and enhance the anticancer efficacy on subcutaneous HepG2-bearing nude mice, which might be due to the enhanced solubility of the drug as well as intracellular delivery (Chapter 2); (2) with BR2 penetrating peptide modification, the liposomal CTD can get into cancerous cells specifically and penetrate deeper in 3D tumor models. A better tumor growth inhibition was also seen in the subcutaneous HCC mice of BR2-modified liposomes treatment than that of the other group, which could be contributed to the passive targeting of liposomes as well as the specific penetrating properties induced by BR2 peptide (Chapter 3); (3) the dual-functionalized liposomes with BR2 peptide and anti-CA IX antibody modification can enhance the drug internalization into HepG2 cells and further improve the anticancer efficacy of drugs compared to other formulations on orthotopic HCC nude mice (Chapter 4). Conclusion: These results demonstrate 1) the liposomal delivery system as a powerful tool to improve anticancer effects of chemotherapeutic agent; 2) the usefulness of BR2 and CA IX modified-liposomal nano-delivery of CTD and their combination might be a potential modality for HCC treatment. The study paved a way for clinical translational medicine of this ligands-modified liposomal delivery system for targeted treatment of HCC.

Aux premières stupeurs de deux morts si rapides que rien ne faisait présager, succèdent les suppositions. Les langues se délient, les réflexions s'échangent. On ne dissimule plus ce que l'on pense et on ne tarde pas à prononcer le mot « empoisonnement ». Le parquet hésite et devant l'insistance de la rumeur publique, il cède. Les trois experts rouennais sont unanimes : sans pouvoir identifier avec certitude le poison utilisé, les sieurs Druaux et Delacroix sont bien morts empoisonnés. L'hypothèse la plus probable est celle des cantharides. La veuve Druaux, restée seule avec les deux victimes, est désignée coupable le 15 novembre 1887 d'avoir tué son mari et son frère à Malaunay, en Seine-Inférieure. Comment, neuf ans plus tard, le procès pourra-t-il faire l'objet d'une révision ? La réponse était dans le nom même de leur habitation : « La maison du four à chaux ». Le coupable est un gaz : le monoxyde de carbone. Incolore, inodore, probablement l'un des poisons des plus subtils et des plus puissants. Il aura fallu trois morts et de nombreux accidents pour l'incriminer et libérer une âme innocente. Les experts parisiens s'intéressent à cette affaire et établissent une violente contre-expertise. Plus qu'un prétexte, cette affaire est véritablement l'occasion de faire un point sur les connaissances médicales et scientifiques à la fin du XIXe siècle sur les cantharides et l'oxyde de carbone. Une mise en perspective avec des éléments plus actuels et les progrès faits depuis permet de donner du sens aux acquis de l'époque.

碩士
臺北醫學大學
藥學研究所
95
Mylabris phalerata, Meloidae family of Coleoptera, has been known since antiquity to produce a potent toxic defensive agent, now known as cantharidin （exo,exo-2,3-dimethyl-7-oxabicyclo〔2.2.1〕heptane-2,3-dicarboxylic anhydride), but more infamously known as Spanish Fly. In the past, Cantharidin has used as anticancer, herbicidal, pesticide, hair-growth stimulator and treatment of molluscum contagiosum activities. In order to study the cytotoxic effects of related compounds, we synthesized four series of cantharidinimides and cantharidinimines by the following two methods：Cantharidin with various primary amines were not only heated to 200℃ in miui-reactor for two hours, but also heated to 140℃ in the miui-reactor for four hours. Furthermore, because of proving the latter structure we used catharidinimine and sodium borohydride in the reductive reaction to obtain amino-containing derivatives. All of the cantharidinimides, cantharidinimines and relative derivatives were identified by 1H-NMR, IR and mass spectrometry, respectively. Moreover, we would try to evaluate cytotoxic effects of cantharidinimide, cantharidinimine and relative derivatives.

碩士
中山醫學大學
應用化學系碩士班
97
Previous studies indicated that cantharidin (exo-2,3-dimethy-7-oxabicyclo [ 2,2,1 ] heptane-2,3-dicarboxylic anhydride)could inhibit the proliferation of cancer cells via p53 dependent mechanism. It could also inhibit protein phosphatases 1(PP1) and protein phosphatases 2A( PP2A ). When cancer cell was treated with cantharidin, it could induce to produce oxidative stress. Oxidative stress would cause increments of single and double stand DNA damage. But the mechanism of anticancer is still not clear. Here, cantharidin and cantharidin derivatives could be served as photo-initiated DNA cleavage agents. We irradiated at λ=352 nm to see whether they have the ability to cleave the super-coiled DNA. The results displayed that the cantharidin derivatives ( 2,6-Dimethyl-4-[4-(6-methyl-benzothiazol-2-yl)-phenyl]-10-oxa-4-aza-tricyclo[5.2.1.02,6]decane-3,5-dione ) could cause DNA scission at acidic condition upon UV irradiation. Then, we used high-resolution gel electrophoresis to analyze the cleavage patterns. The Maxma-Gilbert sequencing data were served as sequence-specific or base-specific cleavage marks. The results indicated that cantharidin derivatives ( 2,6-Dimethyl-4-[4-(6-methyl-benzothiazol-2-yl)-phenyl]-10-oxa-4-aza-tricyclo[5.2.1.02,6]decane-3,5-dione ) have better ability of cleavage at the guanine site.

Aim. The aim of this study was to synthesize and characterize novel analogues of [DACH-Pt-DMC] by using different stereoisomers of DACH; and to investigate any differences in in vitro activity of these complexes in human hepatocellular carcinoma (HCC), colorectal carcinoma (CRC) cell lines and acquired cisplatin or oxaliplatin resistant sub-lines, and to compare that of oxaliplatin and other established Pt-based anticancer agents. Mechanistic roles of DACH-Pt- and DMC components of the TCM-Pt complexes on affecting HCT 116 human CRC cell line were investigated by flow cytometry, COMET assay and cDNA microarray analysis.
Background. Demethylcantharidin (DMC), a modified component of the traditional Chinese medicine (TCM), integrated with a platinum (Pt) moiety created a series of TCM-Pt complexes [Pt(C8H8O 5)(NH2R)2] 1-5 which demonstrated superior antitumor activity and circumvention of cisplatin resistance in vitro. Compound 5, derived from the 1,2-diaminocyclohexane (DACH) ligand (where R=trans-C6H10) had the most potent antitumor activity and closest structural resemblance to oxaliplatin (R,R-DACH-Pt complex) which is the first Pt-based anticancer drug to demonstrate convincing clinical activity against colorectal cancer and has a mechanism of action and resistance that is clearly different from that of cisplatin and carboplatin.
Conclusion. This study is the first to examine the mechanism of anticancer activity of new complexes that integrate DMC with different isomers of DACH. It has shown that both DACH-Pt- and DMC components contribute significantly to the compounds' potent anticancer activity, but likely with different mechanisms of action. The DACH-Pt- component appears to dictate the cell cycle distribution, whereas the DMC component appears to enhance cytotoxicity by inducing more DNA damage in HCT 116 colorectal cancer cells.
Methods. DMC was reacted with appropriate DACH-Pt-(NO3) 2 intermediates, which were prepared from treatment of K2PtCl 4 with stereoisomeric DACH (RR-, SS- & cis-), followed by reaction with silver nitrate. Proton NMR, high-resolution MS, polarimetry and circular dichroism (CD) spectroscopy were used to characterize their chemical structures and optical activities. In vitro antitumor activity (IC50 of 72hr drug exposure time) were assessed by a standard MTT assay. Cell cycle analysis by flow cytometry was determined at 0, 6, 12, 18, 24, 48 and 72 h after drug treatment (cisplatin, carboplatin, oxaliplatin, DMC, compound 1 or trans-DACH-Pt-DMC analogues) at IC50 and 5 x IC50 concentrations with three to four replicates. Comet assay was performed with a fluorescent microscope and used to examine DNA damage after drug treatments (50muM of cisplatin, carboplatin, oxaliplatin, DMC, compound 1 or R,R-DACH-Pt-DMC) for 3hr. cDNA microarray was performed on Affymetrix Human Genome U133A Set and used to analyze gene expression profiles in HCT 116 exposed to trans-(+/-)-DACH-Pt-DMC or oxaliplatin at their IC50 for 72hr.
Results. The in vitro results showed that the trans-analogues were consistently the most potent amongst all the compounds tested in both HCC and CRC cell lines: the trans-(+)(1R,2R)-DACH-Pt-DMC complex, in particular, was the most effective stereoisomer. All of the stereoisomeric DACH-Pt-DMC complexes and oxaliplatin were apparently able to circumvent cisplatin resistance in Huh-7 and SK-Hep1 sub-lines, but cross resistant with oxaliplatin in HCT 116 oxaliplatin resistant sub-line. Flow cytometric analysis revealed the novel trans-DACH-Pt-DMC analogues and oxaliplatin behaved similarly: that is, the compounds at 5 x IC50 concentrations all caused a significant decrease in the S-phase population within 18h and at the same time induced G2/M arrest, and without obvious sub-G 1 phase accumulation, but distinct from that of cisplatin, carboplatin or DMC. Comet assay showed that trans-(+)-(1R,2 R)-DACH-Pt-DMC caused the most significant DNA damage at an equivalent molar concentration. Microarray analysis suggested that the mechanistic role of the DMC ligand can induce the cell cycle to accelerate from the G 1 to S-phase and cause M-phase arrest.
Yu Chun Wing.
"July 2006."
Advisers: Yee-ping Ho; Chik Fun Steve Au-Yeung.
Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1586.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2006.
Includes bibliographical references (p. 191-232).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.

碩士
元培科學技術學院
影像醫學研究所
95
The main extraction method is the traditional solvent extraction. The traditional method is 24 hours in extraction time and must utilize a variety of solvents. The obtained fractions were analyzed by reversed-phase HPLC. The experiments were carried out in the pressure range of 5-100 atm, and modifier volume of 2-6 mL, the temperature range of 25-70 °C, the static extraction time of 5-40 min,. This experiment is utilized mainly Soxhlet extraction, High pressure solvent extraction, High pressure water extraction, Sonication vibration and subcritical fluid CO2 modified with acetonitrile. An experimental design was carried out to map the effects of different parameters on the extraction ability of cantharidin and to determine the optimal conditions for the extraction of cantharidin from Mylabris. The Soxhlet extraction using methanol as a solvent performed for 24 h was 7.35 mg/g, and using acetonitrile as a solvent performed for 24 hrs was 9.49 mg/g. The high pressurized acetonitrile extraction as solvent performed for 30min at 5 atm and 50 °C was 8.39 mg/g. The Sonication vibration extraction was 7.18mg/g.The extraction by subcritical fluid CO2 modified with acetonitrile after a 30-min extraction at 10 atm and 60°C was 9.18 mg/g, The results indicated the CO2 fluid method is definitely may substitute conventional extraction method for cantharidin in the Mylabris.

碩士
國立臺灣大學
獸醫學系研究所
85
The pathophysiologic effects of cantharidin (Can) and norcantharidin (NC) in d ogs were evaluated and compared in this study. Twenty-four dogs were divided i nto two groups and subdivided into eight subgroups. Can control (Can-cont) sub group was administered acetone only, that was the solvent of Can, at 0.1 ml/kg . Can low, median and high doses (Can- L, M and H) subgroups were administered Can with a dose of 0.5, 1.0, 2.0 mg/kg respectively. NC control (NC-cont) sub group was administered normal saline only, that was the solvent of NC, with a dose of 0.4 ml/kg. NC low, median and high doses (NC-L, M and H) subgroups wer e administered NC with a dose of 1.25, 2.5, 5.0 mg/kg respectively. Each dose was administered intravenously. The general body condition (spirit, appetite, body temperature, urination, defecation, hydration status), hematology (RBC co unt, hemoglobin, hematocrit, WBC count and differential count, platelet count) , serum biochemistry (aspartate aminotransferase, alanine aminotransferase, al kaline phosphatase, creatine phosphokinase, blood urea nitrogen, creatinine, t otal protein, albumin, glucose, cholesterol, triglyceride, amylase), urinalysi s (strip method), blood pressure, serum cortisol and testosterone concerntrati on were evaluated pre-administration and post-administration for 3 days. In ad dition, the dogs which died of high dose Can or NC were examined by necropsy a nd histopathology. No significant differences of those evaluated parameters me ntioned above were found in NC-L and Can-L subgroups, besides of rinsing serum creatine phosphokinase activity in Can-L subgroup. There were general body co ndition changes, leukocytosis, thrombocytopenia, liver and kidney damage, card iac muscle damage, lower urinary tract and alimentary tract damage and serum t estosterone decrease in Can-M and NC-M subgroups. Among those, the clinical si gns of the Can-M subgroup were worse than those of the NC-M subgroup. The clin ical signs mentioned above were much more severe in Can-H and NC-H subgroups. Except that, the dogs developed circulatory collapse and died around 6 hours a fter administration. According to the results obtained, the commendatory drug for the cancer treatment trial is NC, and the recommended dose is 0.4-0.6 mg/k g. The dose could be increased in need, but one should pay attention to the to xicity that the dose is over 1.25 mg/kg. Beware of the dose over 2.5 mg/kg as it may cause the patient's death.

碩士
台北醫學院
藥學研究所
89
Cantharidin(exo,exo-2,3-Dimethyl-7-oxabicyclo[2,2,1]heptane-2,3-dicar- boxylic anhydride ) is an active principle of Lytta caragarae, Mylabris Phalerata and Epicauta gorhami, and exists in various other insect species. We used cantharidin as a starting material by reaction with primary amine ex: diaminobenzene, diaminopyridine, ethanolamine and their derivatives and heating ca. 200°C to give various cantharidinimide derivatives. The two amino groups attaching to a benzene ring had three kinds of position isomers ortho, meta, and para. After dehydration and cyclization, cantharidin was converted to cantharidinimides. The inductive effect, resonance effect and steric hindrance effect might reflect the yields. The formation of imide group had electron-withdrawing character influence on benzene ring and displayed a meta activating effect. Furthermore we used some diaminobenzene with some functional groups, for example, the electron-withdrawing group, NO2 , the electron-donating group, OH, and the methyl group. These functional groups might exert the electron-withdrawing and electron-donating capability with resonance and displayed the different basicity to influence the reaction. Because the results obtained with diaminobenzene derivatives, strongly confirmed the influence of amine nucleophilicity and basicity compared the imide group with other functional groups on the benzene ring. We described that the nitro group is a stronger electron withdrawing and meta activating characters than the imide group, the amino and hydroxy are both electron donating groups and had ortho, para activating characters. The methyl group might have a steric hindrance effect in this reaction. Our results indicated that methyl group at the ortho position of the amino group on the benzene ring, the steric hindrance effect became stronger. Pyridine was a heterocycle compound with one nitrogen atom, and nitrogen atom was more electronegative than the carbon atom, the yields were low. In contrast, the long pair of the nitrogen might increase the electron density at the second and fourth position on pyridine ring via the resonance effect. Ethanolamine derivatives were aliphatic compounds, so the yields of cantharidinimide derivatives were only influenced by steric hindrance effect. The more steric hindrance resulted in the less yields. All of the cantharidinimide and their derivatives were measured by 1H-NMR, IR, mass spectrometry. Besides, the series of synthetic compounds were tested for their capability to suppress growth of human hepatocellular carcinoma cell line Hep 3B and Hep G2, and myeloid leukemia cell line HL-60. The results showed compound 20, N-(2-amino- 3-nitrophenyl)cantharidinimide, exhibited more potent than others, but it was still less effective than cantharidin. Key words : cantharidin, cantharidinimide, antitumor agent

碩士
中山醫學大學
應用化學系碩士班
97
壹、斑螫素類似物的合成及其抗癌活性 Cantharidin isolated from Mylabris phalerata Pallas and other insects is used traditionally as an anticancer drug especially on hepato- cellular carcinoma and leukaemia. But the highly renel toxicity of cantharidin limits its use.In order to improve the toxicity of cantharidin, we use cyclic acid anhydride and primary amines to synthesize 5-Phth- alimidopyrimidine-2,4 (1H, 3H)-dione by reflux method and 5-(5-methyl- thiazol-2-ylimino)-4-oxa-tricyclo [5.2.2.02,6]undec-8- en-3-one by solv- enthermal method. Except for the two synthesied compounds , we bought another four compounds (5-aminouracil, 6-aminouracil, Endo-5-norbornene-2,3-dicar- boxylic anhydride, 4-4''-oxydiphthalic anhydride). Moreover, we use MTT assay and DAPI stain assay to evaluate the cytotoxicity and cell apoptosis of these compounds on SK-Hep1 and Hep G2 and rat heaptocyte. 貳、以維他命 C 及溶熱法合成及鑑定包裹奈米鈰顆粒的碳球 We synthesized the carbon spheres by using hydrothermal (solvent- hermal) method at 160 - 180 ℃. In the experiment, we useVitamin C as carbon sources to react with Ce metal complex. The resulting products were characterized by using scanning electron microscopy (SEM) , X-ray powder diffractometer (XRD) , FT-IR spectrum analysis (FT-IR) and NMR analysis. The SEM results showed that the carbon spheres are in perfect sph- erical morphology and dispersed uniform.The powder XRD results showed that we use vitamin C as carbon sources in solventhermal condition to synthesize carbon spheres can encapsulate Ce metal nanopa- rticles. The FT-IR results showed that the carbon spheres encapsulate Ce metal nanoparticles possess specially functional group, including –OH, C=C, C=O. The 1H NMR results showed that the carbon spheres encapsulate Ce metal nanoparticles possess -CH2OH functional group from vitamin C.

碩士
中山醫學大學
應用化學系碩士班
98
Previous studies have found that cantharidin 1, norcantharidin 2 and its derivatives have anti-cancer activity. Some reports indicate that the inhibition of serine/threonine protein phosphatase has an inhibitory effect in cancer cell lines, including HL60, HT29 and L12102. The purpose of this study, using human hepatocellular carcinoma cells (HepG2, SK-Hep1) and primary culture of normal mouse liver cells (normal rat hepatocytes) as a cell model, is to test the anti-cancer activity of a series of anhydride modified cantharidin derivatives (compound3,compound4 and compound 5). The results show that these derivatives can lead to apoptosis of human hepatocellular carcinoma cells (HepG2, SK-Hep1) ,but that in the primary culture of rat hepatocyte is not observed. Their half-death rate (IC50) results in liver cancer cell lines and primary culture of normal mouse liver cells as a cell model for compound1-5 showed that in liver cancer cell line (HepG2) values were 6.2 (μM) ,19.86 (μM) ,150.65 (μM) ,199.18 (μM) ,225.67 (μM) respectively; in liver cancer cell line (SK-Hep1) values were7.79 (μM) ,19.9 (μM) ,21.66 (μM) ,21.02 (μM) ,20.22 (μM) , respectively; in primary culture rat liver cell values were 8.43 (μM) ,35.51 (μM) ,253.34 (μM) ,405.86 (μM) , 262.03 (μM) (Table 1) , respectively, respectively. Therefore, this study tells us that compound 3, compound 4 and compound 5 may be effective anti-cancer drugs.

博士
中山醫學大學
醫學研究所
98
The anticancer activities of cantharidin, norcantharidin and their analogues have been discussed by suppressing the activity of serine/threonine protein phosphatases. The thiazole derivatives are considered as a kinds of antibiotic which can induce apoptosis in human cancer cells. Synthetic thiazolylimide analogues and anhydride analogues were screened and test for their anticancer activities and cytotoxic effects on human hepatocellular carcinoma cell (HCC) lines HepG2, SK-Hep1 and primary cultured rat hepatocytes. Experimental deta had shown that cantharidin, norcantharidin and their analogues displayed an anticancer effect due to their inhibition activities on these cell lines (HL60, HT29, L1210, etc.). However, cantharidin cannot be used as an anticancer reagent because of its high cytotoxicity in vitro (IC50 = 21μM in primary cultured rat hepatocytes). Synthetic cantharidin analogues with aminothiazole structure 3-9 and anhydride structure 10-12 were screened and tested for anticancer activities and cytotoxic effects on human hepatocellular carcinoma cell (HCC) lines HepG2, Sk-Hep1 and primary cultured rat hepatocytes. From the results of our experiments, We observed compounds 3-9 did not perform as expected, with regard to anticancer activity and they exhibited lower cytotoxicity. Compound 10 promoted apoptosis in HepG2 (IC50 = 62μM) and SK-Hep1 (IC50 = 151μM) cell lines. Compounds 11 and 12 had a similar anticancer potentiality to compound 10. After treatment with compounds 3-12 no cytotoxicity reaction was observed in primary cultured rat hepatocytes (IC50 > 200μM). In order to investigate and study the structure activity relationship (SAR) of these analogues, we suggest that anhydride ether oxygen such as in compounds 1,2,10-12 may be correlated with HCC survival and the elimination of bridging ether oxygen on the ring such as 10-12 can decrease their cytotoxicities.

Research Doctorate - Doctor of Philosophy (PhD)
Treating cancer by targeting protein phosphatases, namely protein phosphatase 1 (PP1) and protein phosphatase 2A (PP2A) is a novel ‘fighting fire with fire’ strategy. There are numerous small molecule inhibitors known to achieve this. Most important to this study, cantharidin, and its demethylated analogue, norcantharidin, offer the simplest structure for subsequent modification. The added benefit of effective membrane permeability, makes these compounds ideal candidates for further development. In contrast to most other anticancer drugs, these compounds stimulate the production of white blood cells by bone marrow, while other anticancer drugs that have the unwanted side effect of inducing myelosuppression. Cantharidin displays kidney toxicity which has prevented its use in mainstream oncology. However, norcantharidin is void of kidney toxicity allowing its development for the treatment of cancer. These biologically active compounds have been shown to have multiple uses such as the treatment of warts. Norcantharidin analogues have also been shown to display anti-parasitic activity against nematode Haemonchus contortus, the barbers pole worm, an intestinal parasite that affects livestock industries. Preliminary analysis of a norcantharidin derivative with a single reduced carbonyl group displayed selectivity towards HT29 (colon; GI₅₀ = 14μM) and SJ-G2 (glioblastoma; GI₅₀ = 15μM) when tested against the NCI 60 cell line panel. Intrigued by this finding, multiple small focused libraries were synthesised and assessed in order to compile structure activity relationships (SAR). Interestingly an analogue with an isopropyl ether showed promise with strong selectivity towards HT29 (colon; GI₅₀ = 19μM) and SJ-G2 (glioblastoma; GI₅₀ = 21μM) cell lines but completely void of activity (>100μM) against all seven remaining carcinoma cell lines tested. Norcantharidin analogues were also tested for anti-parasitic activity against Haemonchus contortus, the barbers pole worm, with multiple analogues displayed activity against Haemonchus contortus with associated LD₅₀_s between 25-40 μM. The observed hit-rate of 5.6% associated with this screening of norcantharidin analogues is far higher than conventionally used drug screening methods usually employed. As part of a toxicity pre-filter, all new antiparasitic compounds are screened against a panel of ten cancer cell lines to ensure the end user was not subjected to toxic compounds being applied in a non-ideal environment such as farming communities. Surprisingly, analogues from a small acrylonitrile library, originally used as an internal standard, displayed high levels of cytotoxicity. Subsequent focused library development based on the acrylonitrile scaffold produced multiple broad spectrum cytotoxic compounds with average GI₅₀ values of 1.1-2.1 μM across the nine carcinoma cell lines examined. Interestingly, some acrylonitrile compounds showed a high degree of specificity towards MCF-7 (breast carcinoma) cells of up to 543 fold over the other carcinoma cell lines tested. Some of these compounds were further shown to selectively target estrogen dependent MCF-7 cells that over express the estrogen receptor (ER+ve) over estrogen receptor negative carcinoma (MDA-MB231) and non-malignant breast tissue (MCF10A) up to 268 and 126 fold respectively. With the high throughput of synthesised analogues, flow chemistry methodologies were developed in order to alleviate some of the associated issues with synthetic medicinal chemistry such as reproducibility between batches and difficulty in scale up for live animal studies. Along with effectively producing specific acrylonitrile derivatives in high purity and yield, these procedures were further developed in other projects leading to the discovery of the most potent protein phosphatase inhibitors developed within the research group.

碩士
臺北醫學大學
藥學系
92
Mylabris phalerata, Meloidae family of Coleoptera, has been known since antiquity to produce a potent toxic defensive agent, now known as cantharidin (exo,exo-2,3-dimethyl-7-oxabicyclo [2.2.1] heptane-2,3- dicarboxylic anhydride), but more infamously known as Spanish Fly. In the past, Cantharidin has used as anticancer, herbicidal, pesticide and treatment of molluscum contagiosum activities. The new analogues contain various amino acids attached to the carboxylic anhydride exhibit antibacterial activity. The compounds with triazole structure have been evaluated their antitubercular activity and the new adamantanyl derivatives have found to have antibacterial activity. In addition, some new 1,3,4-thiadiazoles containing isomeric pyridyl have been found to be active against both Gram-positive and Gram-negative bacteria. We used cantharidin as a starting material to react with primary amine, series of amino acid, diaminotriazole and aminoadamantane, by heating 200℃ in sealed tube to form various cantharidinimide derivatives. Furthermore, we used anhydrides and aminothiadiazole in the same condition to obtain thiazolylimide derivatives. We suppose that it might have some steric hindrance between bulky amino acids and 2,3-dimethyl group of cantharidin and gave low yields. And we found that higher polarity of amino acids didn’t react with cantharidin. All of the cantharidinimide and thiazolylimide derivatives were measured by 1H-NMR, IR and mass spectrometry. Therefore, we would try to evaluate antibacterial activity of cantharidinimide and thiazolylimide derivatives. The results indicated that compound 13 (4-Anthracen-2-yl-2,6-dimethyl-10-oxa-4-aza-tricyclo[5.2.1.02,6]decane-3,5-dione) exhibits higher inhibitory activity against Staphylococcus and Bacillus than others, but is still less effective than penicillin G.

Wang Xin Ning.
"May 2002."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (p. 201-236).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

碩士
臺北醫學大學
生物醫學材料研究所
91
Physical stability of liposomes plays the critical roles in various stages of the applications and development of liposomes. In this study, we applied low molecular weight chitosan to modify the liposomes surface, and investigated the physical stability of liposomes, Moreover, we encapsulated cantharidin to study the effects of chitosan which cause drug leakage. In this study, the liposomes were prepared by the extrusion method. Chitosan was added to liposome dispersion to obtain the mixtures of various compositions. We showed that chitosan decreased zeta potential, and increased the size of liposomes. Whereas, the addition of 1wt﹪chitosan had increased the size of liposome at 25℃, i.e. the behavior of fusion. Also, chitosan would increase the turbidity. Further, the DSC (differential scanning calorimeter) revealed that the addition of chitosan caused no difference in the intravesicle interaction. To understand the behavior of cantharidin release, the conventional liposomes and modifier liposomes, revealed that there were no significant difference in size and the rate of drug release at 25℃ and 4℃. Shear forces (shear rate in 300sec-1 、800sec-1) studies of the conventional liposomes and modifier liposomes showed that there were no significant difference in sizes and the rate of drug release at 37℃. Furthermore, both the conventional liposomes and modifier liposomes showed the rate of drug release was higher at shear rate in 800sec-1 than shear rate in 300sec-1.

碩士
中國醫藥大學
藥學系
96
The past 26 years, the cancer has been the first of ten major causes of the death all the time, and good countermeasure does not treat the cancer yet. Colorectal cancer (CRC) is a common and lethal malignant disease, it’s the second most frequent cause of cancer-related death in the United States and occupy the third place in ten major causes of the death of the cancer in Taiwan. Surgical resection、chemotherapy and radiotherapy is the major treatment modality for CRC, but therapeutical effect and prognosis are not perfect. Therefore, investigate an effective and fewer side effect anticancer druge is the task of top priority. Apoptosis is one of the organism maintains the balanced and will not cause inflammation, it has fewer side effect for humen body. Therefore the most important strategy of chemotherapy is utilize medicine to induce cancer cell apoptosis. Cantharidin is the extract of the blister beetle, Mylabris, has been demonstrated to possess antitumor、Immune adjustment、antibiotic and antivirus effect. This thesis main research purpose to probe into that cantharidin can inhibit grow and anticancer mechanism for colorectal cell line (COLO 205). We use the microscope detects the change of cell type and showing that COLO 205 cell line deal with cantharidin, reduce the cell number、shrink and look like has apoptosome. The MTT assay show that COLO 205 cell line deal with cantharidin, cell survival rate show a dose- and time-dependent manner, IC50 is 20.53μM. Cell cycle analyze show cantharidin induced COLO 205 cell line that cell cycle arrest at G2/M phase then induce apoptosis. Western blot and CDK1 activity show cantharidin can reduce cyclin B and CDK1 activity, and can promote chk1 and p21. The TUNEL、DAPI and annexin-v assay is to display the apoptosis of cantharidin on COLO 205. The MMP and ROS assay show reduce the mitochondria membrane potential and increase reactive oxygen species. Western blot and caspase activity assay demonstrate that cytochrome c、Bax、Bad and Fas are increase, Bcl-2 is decrease,and promote caspase-3、-8 and -9 activity. Comprehensive the above result, cantharidin can inhibited cell cycle and activity mitochodria/stress pathway、death receptor pathway then induced cell apoptosis.

碩士
中國醫藥大學
生物科技學系碩士班
100
Cantharidin (CTD) is one of the component from a traditional Chinese medicine named mylabris, and it is a type of terpenoid. It has been demonstrated that CTD possesses antitumor, antibiotic, antivirus and promotes immunity activities. Many studies have reported that CTD induced membrane blebbing, caspase activation, DNA fragmentation, cell cycle arrest in many human cancer cell lines. However, there is no report showing CTD induced apoptosis in human epithelial carcinoma A431 cells. In this study, we investigated the molecular mechanisms of apoptosis in A431 cells after exposed to CTD. We found that CTD decreased the percentage of cell viability, increased the levels of reactive oxygen species (ROS) and calcium (Ca2+) productions, decreased the level of mitochondrial membrane potential (ΔΨm) and triggered apoptosis in A431 cells. CTD increased the protein levels of cell death pathway such as the Caspase-3, -8, -9, PARP active form, cytochrome c and Bax, decreased the anti-apoptosis protein levels such as Mcl-1 and Bcl-2. Annexin V affinity assay was performed by flow cytometry and our results indicated that CTD significantly induced apoptosis in A431 cells in a time-dependent manner and it promoted the protein levels of JNK-mediated c-Jun signaling pathway. We used the specific caspase inhibitors to pre-treat A431 cells, and then exposed to CTD and results showed that CTD increased percentage of viable cells when compared to CTD treatment alone in A431 cells. Based on those observations, we suggest that CTD can induce apoptosis in A431 cells through the activation of caspase-dependent signaling pathways.

碩士
中國醫藥學院
醫學研究所
88
Chronic hepatitis B virus is one of the most widespread disease in Asia and Africa. Especially in Taiwan, there is having prevalence of hepatitis B carriers. The epidemiological studies showed that the indent rate of hepatoma in those carriers is far beyond the normal population. Another finding showed that a majority of the hepatoma cell line contains hepatitis B virus DNA, hence a high correlation in between them is expected. We have tried to trace the affective components of hepatoma cell line and hepatitis B virus in Mylabris. Cantharidin, the active constituent of mylabris, has been demonstrated to possess antitumor properties, and causes leukocytosis and hemorrhagic property. The pharmacological mechanism is still unknown. Our purpose is to evaluate the effect of cantharidin analogues on hepatoma cell line and hepatitis B virus. The drug concentration caused 50% cellular cytotoxicity in 2.2.15 cell number (CC50) of cantharidin analogues, Na CTD (disodium cantharidate) =3.69mM, Na DHNCTD (disodium dehydronorcantharidate)＞50mM, NCTD (norcantharidin) =14.12mM, DHNCTD (dehydronorcantharidin) =16.61mM, respectively. By Wright-Giemsa stain, the morphology in 12.5mM Na CTD for 12days can observed vacuolization and degeneration changes of the 2.2.15 cell line. By the Fluorescene- activated cell sorting (FACS) analysis of DNA profiles in 2.2.15 cell line, the distribution of cell cycle phase induced that high dose of Na CTD and NCTD inhibited the S phase. The antiviral effects of cantharidin analogues were measured by analysis of HBsAg contents. The HBsAg of PLC/PRF/5 cells in culture medium were measure by enzyme immunoassay (EIA) kits. Cell numbers were determined by trypan blue exclusion. The selective index (SI= CC50/ IC50) sequentially decreased from NCTD＞Na DHNCTD＞DHNCTD＞Na CTD. The experiment revealed that the amounts of HBsAg were decreased by a dose-dependent manner for each drug. In this study, we describe the effect of hepatoma cell line and antiviral activity of cantharidin analogues. These compounds should be considered for development as anti-cancer and anti-HBV drugs.

Ng, Po Yan.
Thesis submitted in: November 2006.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2007.
Includes bibliographical references (leaves 109-120).
Abstracts in English and Chinese.
Acknowledgement --- p.i
Abstract --- p.ii
摘要 --- p.iii
Table of Contents --- p.iv
List of Figures --- p.viii
List of Tables --- p.xi
List of Abbreviation --- p.xii
Chapter Chapter 1 --- Introduction
Chapter 1.1 --- A General Introduction to the Development and Clinical Activities of Platinum Drugs --- p.1
Chapter 1.1.1 --- Platinum Drugs used in a Clinical Setting --- p.4
Chapter 1.1.2 --- Platinum Drugs under Clinical Trials --- p.5
Chapter 1.1.3 --- Platinum Compounds with Dual Mechanisms --- p.7
Chapter 1.2 --- Platinum Drug Antitumor Mechanism --- p.9
Chapter 1.3 --- Limitations of Platinum Drugs --- p.12
Chapter 1.3.1 --- Toxicity --- p.12
Chapter 1.3.2 --- Drug Resistance or Cross Resistance --- p.15
Chapter 1.3.2.1 --- Reduced Drug Accumulation or Increased Drug Efflux --- p.16
Chapter 1.3.2.2 --- Drug Inactivation --- p.18
Chapter 1.3.2.3 --- Enhanced DNA Repair --- p.19
Chapter 1.4 --- Why Combinational Therapy? --- p.21
Chapter 1.4.1 --- Antimetabolites --- p.20
Chapter 1.4.2 --- Topoisomerase Inhibitors --- p.22
Chapter 1.4.3 --- Tubulin-Active Antimitotic Agents --- p.24
Chapter 1.4.4 --- Demethylcantharidin as a potential candidate for drug combination --- p.28
Chapter 1.5 --- Study Objectives --- p.31
Chapter Chapter 2 --- Materials and Methods
Chapter 2.1 --- Cell Lines --- p.33
Chapter 2.2 --- Cancer Cell Preparation
Chapter 2.2.1 --- Chemicals and Reagents --- p.33
Chapter 2.2.2 --- Cell Culture Practice --- p.34
Chapter 2.2.2.1 --- Subcultures --- p.35
Chapter 2.2.2.2 --- Cryopreservation --- p.37
Chapter 2.2.2.3 --- Thawing Cryopreservated Cells --- p.38
Chapter 2.2.3 --- Development of Drug-Resistant Cell Lines --- p.39
Chapter 2.3 --- Growth Inhibition Assay
Chapter 2.3.1 --- Evaluation of Cytotoxicity in vitro --- p.40
Chapter 2.3.2 --- Drug Pretreatment --- p.43
Chapter 2.3.3 --- Drug Pre-sensitization with Concurrent Treatment --- p.44
Chapter 2.4 --- Calculations for Drug Combinations --- p.46
Chapter 2.5 --- Statistical Analysis --- p.49
Chapter Chapter 3 --- Results and Discussions
Chapter 3.1 --- In vitro Cytotoxicity and Evaluation of Drug Resistance --- p.50
Chapter 3.2 --- Role of Leaving Ligand in a Platinum Complex --- p.58
Chapter 3.3 --- Priority in Selecting the Most Effective Drug Combination --- p.66
Chapter 3.4 --- Drug Combination Studies
Chapter 3.4.1 --- Drug Combination Prescreening --- p.68
Chapter 3.4.1.1 --- Comparison of the effectiveness of the three Drug Combinations --- p.72
Chapter 3.4.1.2 --- Rationale for Drug Combination Studies presented in Section 3.4.2 & 3.4.3 --- p.73
Chapter 3.4.2 --- Drug Pre-sensitization Studies in Colorectal Cancer Cell Lines --- p.74
Chapter 3.4.2.1 --- Comparison of Drug Pre-sensitization Treatment in Sensitive Colorectal Cancer Cell Lines --- p.84
Chapter 3.4.2.2 --- Comparison of Drug Pre-sensitization Treatment in Sensitive and Oxaliplatin Resistant HCT116 Colorectal Cancer Cell Lines --- p.87
Chapter 3.4.3 --- Drug Pre-sensitization Studies in Liver Cancer Cell Lines --- p.89
Chapter 3.4.3.1 --- Comparison of Drug Pre-sensitization Treatment in Sensitive Liver Cancer Cell Lines --- p.99
Chapter 3.4.3.2 --- Comparison of Drug Pre-sensitization Treatment in Sensitive and Cisplatin Resistant SK-Hepl Liver Cancer Cell Line --- p.101
Chapter 3.5 --- Possible Explanation to the Observed Drug Combination Effect --- p.103
Chapter 3.6 --- General Protocols for Drug Combinations --- p.105
Chapter Chapter 4 --- Conclusions
Reference --- p.109
Appendices --- p.121
Chapter I a. --- "Raw Data of Pre-screening for HCT116 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.122
Chapter I b. --- "Raw Data of Pre-screening for HCT116 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.123
Chapter II a. --- "Raw Data of Pre-screening for SK-Hepl (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.124
Chapter II b. --- "Raw Data of Pre-screening for SK-Hepl ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.125
Chapter III a. i) --- "Isobolograms for HCT116 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.126
Chapter III a. ii) --- "Raw Data for HCT116 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.127
Chapter III b. i) --- "Isobolograms for HCT116 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.128
Chapter III b. ii) --- "Raw Data for HCT116 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.129
Chapter IV a. i) --- "Isobolograms for HCT1160xaR (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.130
Chapter IV a. ii) --- "Raw Data for HCT1160xaR (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.131
Chapter IV b. i) --- "Isobolograms for HCT1160xaR ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.132
Chapter IV b. ii) --- "Raw Data for HCT1160xaR ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.133
Chapter V a. i) --- "Isobolograms for HT29 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.134
Chapter V a. ii) --- "Raw Data for HT29 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.135
Chapter V b. i) --- "Isobolograms for HT29 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.136
Chapter V b. ii) --- "Raw Data for HT29 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.137
Chapter VI a. i) --- Isobolograms for Hep G2 (Cisplatin and [Pt(DMC)(NH3)2]) --- p.138
Chapter VI a. ii) --- Raw Data for Hep G2 (Cisplatin and [Pt(DMC)(NH3)2]) --- p.139
Chapter VI b. i) --- "Isobolograms for Hep G2 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.140
Chapter VI b. ii) --- "Raw Data for Hep G2 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.141
Chapter VII a. i) --- "isobolograms for SK Hep 1 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.142
Chapter VII a. ii) --- "Raw Data for SK Hep 1 (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.143
Chapter VII b.i) --- "Isobolograms for SK Hep 1 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.144
Chapter VII b. ii) --- "Raw Data for SK Hep 1 ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.145
Chapter VIII a. i) --- "Isobolograms for SK Hep ICisR (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.146
Chapter VIII a. ii) --- "Raw Data for SK Hep ICisR (Cisplatin, [Pt(DMC)(NH3)2] and Pt(DMC)(NH2CH3)2])" --- p.147
Chapter VIII b. i) --- "Isobolograms for SK Hep ICisR ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.148
Chapter VIII b. ii) --- "Raw Data for SK Hep ICisR ([Pt(DMC)(R,R-DACH)] and Oxaliplatin)" --- p.149

碩士
中國醫藥大學
營養學系碩士班
97
Cantharidin derived from traditional Chinese medicine, the blister beetle, Mylabris phalerata, and it had been shown to inhibit the growth of various cancer cells. Prostate cancer is one of most male malignancy which associated with high mortality. Advanced and metastasis cancer development with the growth of androgen-independent tumor cells results in hormone-refractory disease and leads to lethal death. However, there is no information to address cantharidin-induced cell death in human prostate cancer PC-3 cells. After treatment of PC-3 cells with cantharidin for various times, cell viability were examined by MTT assay and specific cellular morphology changes were observed. In addition, incubation of PC-3 cells with cantharidin led to increase DNA damage which were assessed by Comet assay. Cantharidin induced cell cycle arrest in G2/M phase and apoptosis, by staining with PI and determined by flow cytometry. The apoptotic evidences induced by cantharidin, including chromatin condensation and DNA fragmentation, are examined by DAPI staining and agarose DNA electrophoresis assay. Cantharidin increased cytosolic calcium (Ca2+) concentration and decreased the level of mitochondrial membrane potential (ΔΨm), which were stained with Ca2+ probe (Fluo-3/AM) and dihexyloxacarbocyanine iodide (DiOC6) then were analyzed by flow cytometry. Cantharidin also promoted caspase-3 activity in PC-3 cells. The results implied that cantharidin induced apoptosis through mitochondrial- and caspase-3- dependent pathways and it may be a potential cancer therapeutic agent in androgen-independent prostate cancer in future.

博士
中國醫藥大學
中醫學系博士班
102
Lung cancer is one of the leading cause of death in cancer-related diseases. Cantharidin (CTD) is one of the components of natural mylabris (Mylabris phalerata Pallas) which have been used as drugs in Chinese population. Numerous studies also showed that CTD induced cytotoxic effects on cancer cells. However, there is no report to demonstrate that CTD induced apoptosis in human lung cancer cells. Herein, we investigated the effect of CTD on the cell death via the induction of apoptosis in H460 human lung cancer cells. Results showed that CTD induced apoptosis based on the occurrs of sub-G1 phase and DNA fragmentation. We also found that CTD increased gene expression (mRNA) of caspase-3 and -8. Moreover, increased ROS and Ca2+ productions and decreased the levels of ΔΨm. Western blotting results showed that CTD increased the expression of cleavage caspase-3 and -8, cytochrome c, Bax and AIF but inhibited the levels of Bcl-xl. CTD promoted ER stress associated protein expression such as Grp78, IRE1α, IRE1β, ATF-6α and caspase-4 and it also promoted the expression of Calpain2 and XBP-1 but inhibited Calpain1 that is associated with apoptosis pathways. We also found that cantharidin were treatment of H460 cells can reduced protein levels of ataxia telangiectasia mutated (ATM), breast cancer 1, early onset (BRCA-1), 14-3-3 proteins sigma (14-3-3σ), DNA-dependent serine/threonine protein kinase (DNA-PK), O6-methylguanine-DNA methyltransferase (MGMT) and mediator of DNA damage checkpoint protein 1 (MDC1). Protein translocation of p-p53, p-H2A.X (S140) and MDC1 from cytoplasm to nucleus was induced by cantharidin in NCI-H460 cells. Taken together, the present study showed that cantharidin caused DNA damage and inhibited levels of DNA repair associated proteins. We also using microarray analysis to found that CTD acted on genes associated with DNA damage, cell cycle and apoptotic cell death in human lung cancer NCI-H460 cells. These effects may contribute to cantharidin-induced cell apoptosis in vitro. On the other hand, we investigated the effect of CTD on the cell adhension, migration and invasion of NCI-H460 human lung cancer cells. Results indicated that CTD decreased the percentage of viable cells and inhibit the cell adhension are in a dose-dependent manners. CTD not only inhibit cell migration and invasion are in a dose-dependent manners but also inhibit the enzymatic activities of MMP-2/9 of NCI-H460 cells. Western blotting data showed that CTD decreased the expression of MMP-2/-9, FAK, Rho A, Akt, Erk, p38, Jnk, NF-κB and uPA. Furthermore, confocal laser microscopy also show suppressed the expression of FAK, Rho A and NF-κB in NCI-H460 cells. Based on those observations, we suggest that CTD may be used as a novel anti-cancer metastasis of lung cancer.