The Impact Of Interference Xiap Gene On Human Cholangiocarcinoma Qbc939 Cell Proliferation Inhibition And Sensitivity To Chemotherapy

Objective:To study the inhibition of XIAP(X-linked inhibitor of apoptosis protein) gene expression induced by ASODN(Antisense oligodeoxynucleotide) in QBC939 cell line,and the related proliferation inhibition and apoptotic inhibition of QBC939 cells.To investigate the chemotherapeutic sensitivity change of bile duct cancer cells of QBC939 after XIAP inhibition.Methods:To observe the cell of ASODN transfection in a different time period by Fluorescence microscope,and calculate the rate of transfection.The experiment was divided into 4 groups:ASODN group(Anti-sense group),NSODN group(Nonsense group),Liposome group,Control group,ASODN/NSODN were transfected into QBC939 cell line by Lipofectamine^TM 2000.To measure XIAP mRNA expression by RT-PCR after the transfection; MTT assay was used to assess the cell growth of different concentrations.The changes of cell cycle and apoptosis were detected by Flow cytometry(FCM).5-FU concentration is designed to 4,8,16,32,64,128,256,512,1024μg/ml by relevant literature.MTT assay was used to evaluate the inhibitory effects of QBC939 cells in different concentrations of 5-FU,and to calculate IC₅₀.Then,we divided the experiment into 4 groups:ASODN group,5-FU & ASODN group(5-FA group),5-FU group,Control group.To evaluate the inhibitory effects of QBC939 by MTT assay in different groups,and the apoptosis was detected by Flow cytometry(FCM).Results:ASODN can be transfected into QBC939 cells by Transient transfection,and transfection efficiency is very high,which increased as the concentration was elevated with certain extent,when the concentration of ASODN is 1.11μmol/L,the highest transfection efficiency is 95.9±2.0%at 24h.In contrast with the control group,the gene expression level of XIAP mRNA in ASODN group was decreased(p＜0.05). MTT results showed that five kinds of concentration ASDON can inhibit the QBC939 cell growth,and in 0.28μmol/L to 1.11μmol/L,the inhibitory effects of QBC939 increased as the concentration was elevated with certain extent,and the inhibition ratio was higher than the control group(P＜0.05).In ASODN group,the rate of apoptosis was 10.67±2.25%,and the QBC939 cell in G0/G1(58.47±1.90%) were higher those in control group(p＜0.05).The 5-FU IC₅₀ was 550.22±19.10μg/ml,the inhibition ratio (48h 71.64±0.64%) and apoptosis(62.16±1.18%) was higher than those in control group(p＜0.05).Conlusion:The liposomal transfection of ASODN can down-regulate the expression of XIAP gene,,which can inhibit cell growth and induced apoptosis of bile duct cancer cells,and increase the chemotherapeutic sensitivity of bile duct cancer cells of QBC939 after XIAP inhibition.So it may be the new target for gene therapy in bile duct cancer.