| ObjectiveTo research the effect of y-radiation released from 103Pd radioactive stent on the expression of Fas gene and its relation with apoptosis of cholangiocarcinoma cell through the establishment of human cholangiocarcinoma model.MethodsPoorly differentiated human bile duct adenocarcinoma cell line QBC939 cell culture medium containing 10% fetal bovine serum,100 U/ml penicillin,100 U/ml streptomycin in RPMI-1640 culture flask, placed in constant temperature 37.5℃,5% CO2 concentration, saturated humidity, when the total number of cultured cells by cell counting up to 5 x 107个above, with trypsin digestion, centrifugation to collect the bottle cells. Diluted with culture medium made of 1×107/ml, the cell suspension.The establishment of human cholangiocarcinoma xenografts in nude mice, experimental BALB/c-nu mice were purchased from the Institute of Zoology, Chinese Academy of Medical Sciences, mice age 8 to 10 weeks, weighing 15-20 g, either sex. Take primary transplantation; select more than the maximum tumor diameter of 1.0 cm, body mass greater than 20 g nude 36, male or female, were randomly divided into 103Pd stent group and ordinary stent group,18 in each group. Experimental group 37MBq biliary radioactive stent implantation, the control group implanted bare metal stent implantation, mice were killed 10d after stenting calculated tumor volume (V), the formula V (mm3)= ab2/2. Tumor growth curve diagram; application TUNEL assay apoptosis of cholangiocarcinoma cells, the results to judge:the nucleus of a brown stain by the TUNEL-positive cells, that is, apoptotic cells, high magnification view unit count TUNEL-positive cells within,3 times counting the mean value as the number of apoptotic cells. And calculated apoptosis. Electron microscope observation of cell ultrastructure; on 103Pd stent group and the general support group to HE staining for tumor pathology, verification of its human cholangiocarcinoma homology. Immunohistochemistry to detect Fas gene expression in cholangiocarcinoma cells the situation was brown cytoplasm were positive cell, the cytoplasm no brown staining was negative cells. Positive cells divided 3:Low magnification (×10) 1 cm2 outlook positive cells within total cells<1/3 (＋),1/3 to 2/3 (＋＋),> 2/3 denoted (＋＋＋), no positive cells (a). All measurement data are mean±standard deviation application SPSS 12.0 statistical analysis software, according to data of different situations using different statistical methods:①semi-quantitative data usingχ2 test;②quantitative data using analysis of variance,χ2 test and rank sum test. Test level a= 0.05.ResultsCompare with the control group, the increased speed of the volume of tumor in the experimental group is significantly reduced. The expression level of Fas gene in 103Pd stent group is significantly higher than that in general stent group (P<0.05),and the number of apoptotic cancer cells in 103Pd stent group is also significantly more than that in general stent group (P<0.01).ConclusionThe 103Pd Radioactive stent can induce the cell apoptosis in human cholangio-carcinoma transplanted tumor of nude mouse,inhibit the cell growth of bile duct cancer and may promote the apoptosis of cancer cells by increasing the expression of Fas gene. It may be helpful for the further study of treatment for bile duct cancer using 103Pd Radioactive stent.
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