Stat3反义寡核苷酸对人胃腺癌MKN45细胞相关信号传导影响的研究

Effect of Stat3 antisense oligonucleotide on signal transduction pathway related to human stomach adenocarcinoma MKN45 cell line

目的:通过研究Stat3反义寡核苷酸对人胃腺癌MKN45细胞增生活性的影响,寻求胃癌治疗中相关信号传导途径的新靶点. 方法:Stat3反义寡核苷酸是一种混合骨架寡核苷酸(MBO), 采用脂质体介导的方式将其转染人胃腺癌细胞株MKN45; 通过MTT法观察转染前后对细胞增生状态的影响;凝胶阻滞电泳(EMSA)和Western blot方法观察转染前后Stat3 DNA的结合活性和磷酸化Stat3蛋白表达的变化. 结果:Stat3反义寡核苷酸对MKN45细胞的增生有明显抑制作用;转染反义寡核苷酸后MKN45细胞中Stat3信号的组成性激活水平和磷酸化Stat3蛋白的表达分别下降了50.65%及78.86%. 结论:Stat3反义寡核苷酸能显著抑制MKN45细胞中Stat3 信号的传导,胃癌细胞株中活化的Stat3信号可作为胃癌治疗新的分子靶点.

AIM: To investigate the effect of antisense oligonucleotide (as-ODN) of Stat3 on the cell proliferation of human stomach adenocarcinoma cell line MKN45 and to determine the novel molecular target for treatment of gastric cancer. METHODS: Stat3 as-ODN, a mixed backbone oligonucleic acid (MBO), was transfected into MKN45 cells mediated by liposomal reagent. The effect on cell proliferation was examined by MTT method. After transfection of Stat3 as-ODN, electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the Stat3 DNA-binding activity and the expression of phospho-Stat3 protein, respectively. RESULTS: Stat3 as-ODN could significantly inhibit the proliferation of MKN45 cells. After transfection of Stat3 as-ODN, both the constitutive activation of Stat3 and the expression of phospho-Stat3 protein were decreased 50.65% and 78.86%, respectively. CONCLUSION: Stat3 as-ODN could remarkably inhibit the signal transduction of Stat3 in MKN45 cells. Activated Stat3 signaling in human stomach adenocarcinoma cell line provides a novel molecular target for therapeutic intervention of gastric cancer.