转染反义寡核苷酸对人胆管癌QBC939细胞XIAP基因表达及细胞周期作用的实验研究

目的研究转染反义寡核苷酸（antisense oligodeoxynucleotide，ASODN）对人胆管癌细胞株QBC939细胞凋亡抑制蛋白基因（X-linked inhibitor of apoptosisprotein，XIAP）表达的抑制作用及对细胞周期凋亡的影响．方法脂质体介导ASODN瞬时转染QBC939细胞，荧光显微镜观察ASODN转染入细胞的情况，并计算转染率；RT—PCR检测转染前后XIAPmRNA表达的变化；流式细胞仪检测转染后细胞周期的变化和凋亡率．结果转染后24h可达到最佳的转染效果；ASODN组的XIAP基因水平明显低于对照组（P〈0．05），流式结果显示转染后QBC939G0／G1期细胞高于阴性对照组，凋亡率高于对照组（P〈0．05）．结论脂质体介导转染ASODN能特异性地抑制QBC939细胞中的XIAP基因表达，使其发生G0／G1期阻滞，并诱导胆管癌细胞凋亡，有望成为胆管癌基因治疗新的靶点．

Effect of Transfection of Antisense Oligodeoxynucleotide on IAP Gene Expression and Cell Cycle of QBC939 Cells

Objective To study the inhibition effect of transfection of antisense oligodeoxynucleotide （ASODN） on XIAP （X-linked inhibitor of apoptosis protein） expression of QBC939 cells, and the effect of cell cycle and apoptosis. Methods ASODN were transfected into cell line QBC939 by LipofectamineTM 2000, the cells of ASODN transfection were observed by Fluorescence microscope, and the rate of transfection was calculated; XIAP mRNA expression was detected by RT-PCR after the transfection; The changes of cell cycle and apoptotic were detected by Flow cytometry （FCM）. Results The highest efficiency was：at 24 hours after liposomal transfection. The gene expression level of XIAP in ASODN group was significantly lower than that in control group （P 〈 0.05 ）. The results of FCM showed that the rate of apoptosis and QBC939 cells in the cell cycle of G0/G1 were higher than those in control group （P 〈 0.05）. Conclusions The liposomal transfection of ASODN can inhibit XIAP gene expression, and block QBC939 cell in G0/G1, and induce apoptosis of bile duct cancer cells, which may be the new target for gene therapy in bile duct cancer.