干扰X连锁凋亡抑制蛋白基因对人胆管癌QBC939细胞增殖及凋亡的作用

目的 观察反义寡核苷酸(ASODN)对人胆管癌QBC939细胞株X连锁凋亡抑制蛋白(XIAP)基因的抑制作用及对癌细胞增殖及凋亡的影响.方法 脂质体法将ASODN转染入QBC939细胞中,转染12、24、48 h后荧光显微镜观察转染后细胞状态及转染率,应用噻唑蓝(MTT)比色法、逆转录-聚合酶链反应(RT-PCR)法和流式细胞仪法测定1.11μmol/L时细胞生长抑制率、XIAP mRNA表达和细胞周期及凋亡率.结果 转染效率可达95%;与对照组比较,1.11μumol/L细胞抑制率为(27.63±1.15)%(24 h),(42.95±1.07)%(48 h);mRNA下调23%(P<0.05);G_0/G_1期细胞、凋亡率高于对照组约27.34%、9.94%(P<0.05).结论 ASODN能有效下调QBC939细胞XIAP基因表达,抑制细胞增殖并诱导凋亡.

Effect of XIAP gene interference on proliferation and apoptosis of human QBC939 cells

Objective To study the inhibitory effects of antisense oligodeoxynucleotide (ASODN) on X-linked inhibitor of apoptosis protein (XIAP) gene expression in QBC939 cell line,and the effects on proliferation and apoptosis of QBC939 cells. Methods ASODN was transfected into cell line QBC939 by LipofectamineTM 2000,and transfection efficiency at the 12th,24th,and 48th h after transfec-tion was observed by fluorescence microscopy. Proliferation inhibition rate, XIAP mRNA expression,cell cycle and apoptosis rate of QBC939 cells treated with 1. 11 μmol/L ASODN were assayed by MTT,RT-PCR and flow cytometry (FCM). Results Transfection efficiency was about 95%. In contrast with the control group,the inhibition rate of cell proliferation was about (27.63 ±1.15)% (24 h) and (42.95 ± 1.07) % (48 h) respectively after treatment with 1.11 μmol/L ASODN. The gene expression level of XI-AP in ASODN-transfected group was decreased by 23% (P<0.05). The apoptosis rate and the percent-age of QBC939 cells in G_0/G_1 were increased by 27. 34% and 9. 94% in ASODN-transfected group as compared with control group (P<0. 05). Conclusion The liposomal transfection of ASODN can down-regulate the expression of XIAP gene, which can inhibit proliferation and induce apoptosis of QBC939 cells.