| Cholangiocarcinoma has a poor prognosis among the alimentary duct cancers. Currently, the best therapeutic program that combined resection with chemotherapy or radiotherapy or immunotherapy and others is identified extensively because of the lower resection rate and the poor effect after operation, although resection is considered to be the the most important strategy. Chemotherapy plays an important role in the therapeutic program, but its effect on cholangiocarcinoma is still not ideal.The total efficiency documented in the literature was not superior than 50% with the simplex chemotherapy, additionally the side-effect or poison of drug was seriously harmful to body. Thus, it is necessary to ques new drug ameliorating the chemotherapeutic effect on cholangiocarcinoma.In the past decades, the prognosis of acute promyelocytic leukemia (APL) has been made a rapid progress by the arsenic trioxide, a traditional Chinese medicine, which has a high clinic remission rate and little side-effect. The anticancer mechanism had been demonstrated to inhibit proliferation and produce apoptosis. Recently some experiments also showed arsenic trioxide had effect on hepatocarcinoma, esophageal carcinoma, neuroblastoma and gastric carcinoma in vitro in a similar manner, however, arsenic trioxide had no effect on some cell strain. Thus, our aim is to investigate the effects of As2O3 on human changiocarcinoma QBC939 cell line and to illustrate the reasonable mechanism in this study.The main objects and methods include some aspects as following, (l)Toininvestigate the cell proliferation kinetics by drawing cell growth curve diagram and recording clone formation; (2)To compare the drug susceptibility of AsiOs with DDP, MMC, VCR, VP-16, 5-Fu and ADM in vitro by MTT reduction assay; (3) To detecte apoptosis by Acridine orange fluorescence staining, transmission electron microscope and in situ end-labeling of fragments (TUNEL assay), then analysing cell cycle by FCM.;(4)To measure cell invasive capacity by Boyden chamber system; (5)To investigate apoptosis and proliferation of the human umbilical vein ECV304 cell line in the same way as QBC939 cell line and evaluating the effects of As2O3 on ECV304 proliferation cultured in QBC939 conditioned medium by MTT reduction assay. At the same time, observing angiogenesis on the chicken embryo illus allantois membrane(CAM) ;(6)To establish chicken transplanted cholangiocarcinoma model , then compare tumor size, VEGF and PCNA expression (by SABC) and apoptosis (by TUNEL) between the AS2O3 group and the contra! group.The main results are as follows: (1)0.75 U mol/L~ 12 u mol/L As2(>3 could inhibit QBC939 cell proliferation and clone formation resulting in a reproducible dose- and time- dependent sequence of events marked by change to an activated morphology. (2) ADM, AsaOa and DDP were the relatively sensitive drugs to QBC939 cell line by the drug susceptibility detection in vitro. (3)As2C>3 can induce QBC939 cell apoptosis in a tune- and dose-dependent manner, and 6 n mol/L AsaOs in 72h led to an apoptosis 34.4?.28% ; FCM prompted that AsiOa blocked cell cycle representing S t , G2/M t and GO/G1I .(4)As2O3 can depress QBC939 cell invasiveness and motor ability. (5)Cellular bcl-2> PCNA and uPA (by SABQexpressed down-regulation . (6)AsaO3 can induce ECV304 cell apoptosis and blocke cell proliferation in the same way as QBC939 cell.. QBC939 conditioned medium facilitated ECV304 cell proliferation, whereas As2Os depressed the role .(7) As2O3 inhibited CAM angiogenesis and the growth of transplanted cholangiocarcinomaIVon CAM. Tumor PCNA and VEGF expressed down-regulation, but tumor cell apoptosis increased crossly.At the serum concentration of 4.2 u mol/L~6.7 u mol/L for the treatment of acute promyelocytic leukemia (APL), AS2O3 can significantly inhibit proliferation of the QBC939 cell line and inhibit growth of the transplanted chicken embryo cholangiocarcinoma. AsaOs is a high sensitive drug to QBC939 cell line in comparison with other chemotherapy drugs. |
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