Survivin、xiap、apollon和livin反义寡核苷酸对人消化系肿瘤细胞增殖及凋亡的影响

摘要

Aim To explore the proliferatory inhibition effects of survivin, xiap, apollon and livin of inhibitor of apoptosis proteins family members antisense oligodeoxynucleotide ( ASO ) on gastrointestinal cancer cells.Methods The antisense oligodeoxynucleotide targeting for surviving, xiap, apollon, livin and a random oligodeoxynucleotide were synthesized and transfected into gastrotestinal cancer cells ( HepG2, MGC-803,Lovo ) using LipofectaminerTM 2000 for 48 hours.cell proliferation inhibition was determined by WST method,and cell apoptotic level examined by flow cytometry.Results Antisense oligodeoxynucleotide targeting for surviving, xiap, apollon, livin gene could suppress the growth of gastrointestinal cancer of HepG2, MGC803 , Lovo cells and induce apoptosis of these cells significantly ( P＜ 0.05 ).At a concentration of 0.2 μmol · L-1, survivin ASO had a significantly inhibited effects on the HepG2, Lovo cells.The inhibitory rates were 84.2％ ,82.6％ respectively.Xiap ASO had a significantly inhibited effects on the MGC-803 cell.and the inhibitory rate was 86.56％.They showed time and dose-dependent manners.The effects of survivin ASO's inducing apoptosis on Lovo cell, MGC-803 cell were stronger than other ASOs.The apoptotic rate was 34.95％ , 27.6％ respectively.The inducing apoptosis effect of livin ASO on HepG2 cell was stronger than other ASOs and the cell apoptotic rate was 27.4％.Conclusion Antisense oligodeoxynucleotide targeting IAPs can inhibit the proliferation and induce the apoptosis of gastrointestinal cancer cells in vitro.%目的 观察凋亡抑制蛋白(inhibitor of apoptosis p roteins, IAPs)家族survivin、xiap、apollon和livin反义寡核苷酸(antisense oligodeoxynucleotide,ASO) 对人消化系肿瘤细胞增殖、凋亡的影响.方法 人工合成survivin、xiap、apollon、livin的反义核酸和随机序列,经脂质体包裹后作用肝癌细胞、胃癌细胞、结肠癌细胞48 h后,采用WST-8法观察对细胞增殖的影响,流式细胞术检测细胞凋亡率.结果 IAPs ASO作用于肝癌HepG2细胞、胃癌MGC-803细胞、结肠癌Lovo细胞48 h后,与空白对照组(control)和随机寡核苷酸(random oligodeoxynucleotide, RODN)对照组相比,IAPs ASO能明显抑制肝癌细胞、胃癌细胞、结肠癌细胞的增殖和诱导细胞凋亡(P<0.01).其中,在作用浓度为0.2 μmol·L-1时, survivin ASO对HepG2、Lovo细胞的增殖抑制率最高,分别为84.2%、82.6%.xiap ASO对MGC-803细胞的增殖抑制率最高,为86.56%,并呈时间-剂量依赖效应.survivin ASO诱导Lovo细胞、MGC-803细胞凋亡作用最强,凋亡率为34.95%、27.6%,livin ASO诱导HepG2细胞凋亡作用最强,凋亡率为27.4%.结论 针对凋亡抑制蛋白家族survivin、xiap、apollon、livin的反义核酸均能有效抑制消化系肿瘤细胞增殖,并诱导细胞凋亡.