Studies On The Screening Of Cholangiocarcinoma Drugs And The Mechanisms

Cholangiocarcinoma is a malignant tumor which is difficult to cure.At present,radical surgical removal represents the only curative treatment.However,the tumoroften present at an advanced unresectable stage at the time of diagnosis for the majoritypatients,which eliminates the surgical approach as a curative measure.Thus,it isimportant to find out a effective therapeutic method excluded surgical.The response ofcholangiocarcinoma to current clinical chemotherapeutic regimens is poor.The currentclinical drugs for cholangiocarcinoma therapy often have toxic and side effects.Therefore,to improve the curative effect of drug therapy,it is urgent to find moresensitive and effective therapeutic drugs for cholangiocarcinoma.In this study,cultured human cholangiocarcinoma cell line QBC939 in vitro wasused as the model for drug screening.Extracts from Phyllanthus niruri L.,Phyllanthusurinaria L.,Ilex chinensis sims,Solanum lyratum Thunb.,Catharanthus rosens(L.)G.Don,Cephalotaxus fortunei Hook.f.which are medicinal plants and CorbiculaFluminea which is a kind of freshwater mollusk,natural compounds,thiosemicarbazone compounds and common chemotherapeutic drugs were selected toevaluate the growth inhibitory effects towards cholangiocarcinoma by MTT method.The results showed that ethyl acetate and butanol extrats from Phyllanthus urinariaL.and Phyllanthus niruri L.,water extract from Solanum lyratum Thunb.,Vincaalkaloids,ethyl acetate extrat from Corbicula Fluminea,quercetin,tannic acid,2-chlorobenzaldehyde thiosemicarbazone,2,4-dichlorobenzaldehydethiosemicarbazone and tamoxifen exhibit strong growth inhibitory effects towardcholangiocarcinoma cells.Compared to 5-fluorouracil and cisplatin,tamoxifen was more sensitive tocholangiocarcinoma cells.Therefore,we did futher studies to investigate the antitumormechanisms of tamoxifen (TAM)on cholangiocarcinoma cells.By means of MTTmethod,morphologic observation method,flow cytometry,DNA ladder method andother methods,the results showed that TAM could significantly inhibit the growth ofQBC939 cells in a time-and dose-dependent manner,change the morphology of QBC939 cells,arrest the cell cycle at G₀/G₁ phase and induce apoptosis.By means ofwestern blot and other methods,the results showed that inhibiting the expression ofCyclin D1,ERα,C-Myc,activation Caspase-3/Caspase-9 signal pathway andup-regulating the expression of Bax and p53 were considered to be the partialmechanisms.To understand the possible molecular mechanisms and function targets ofinhibitory effect induced by TAM on cholangiocarcionoma cells,proteomicstechnology was first applied to investigate the effect of TAM on proteomic profile ofQBC939 cells.Heat shock proteins,cytoskeletal proteins,membrane associated protein,metabolic related enzymes and other differential expression proteins were successfullyidentified.According to the functions of these proteins,we analysed the possiblemolecular anticancer mechanisms of TAM on cholangiocarcinoma cells comprehensivecomprehensively and systematically.All our findings provide a new clue for the drugtherapy of cholangiocarcinoma,provide theoretical support for application of TAM oncholangiocarcinoma therapy in clinic and provide new targets and strategies forelucidating the anticancer mechanisms of TAM on cholangiocarcinoma.