ErbB2基因沉默介导PI3K/Akt/eNOS信号通路对卵巢癌细胞生物学特性影响

EFFECT OF ERBB2 GENE SILENCING ON THE BIOLOGICAL CHARACTERISTICS OF OVARIAN CANCER CELLS BY MEDIATING THE PI3K/AKT/ENOS SIGNALING PATHWAY

目的探讨ErbB2基因沉默介导PI3K/Akt/eNOS信号通路对卵巢癌细胞生物学特性的影响及机制。方法选取对数生长期人卵巢癌细胞株HO8910细胞分为5组:空白对照组(不转染任何序列)、阴性对照组(转染空载体质粒)、siErbB2组(转染siRNA序列)、wortmannin组(转染PI3K/Akt/eNOS信号通路抑制剂wortmannin)、siErbB2+IGF-1组(转染siRNA序列+PI3K/Akt/eNOS信号通路激动剂IGF-1)。采用实时定量逆转录聚合酶链反应(qRT-PCR)和Western blot检测ErbB2、Akt、PI3K、eNOS、Caspase-3、Bax和Bcl-2的mRNA和蛋白表达水平,同时检测细胞增殖、凋亡、迁移和侵袭的变化。结果与空白对照组比较,siErbB2组和wortmannin组卵巢癌细胞中Akt、PI3K、eNOS和Bcl-2的mRNA和蛋白表达量显著下降,而Caspase-3、Bax的mRNA和蛋白表达量则显著增加(F=3.58~8.69,P均<0.05),细胞增殖能力、侵袭能力、迁移能力均显著下降(F=8.84~15.67,P均<0.05),细胞凋亡率上升(F=9.46,P<0.05);siErbB2组ErbB2表达明显下降(P<0.05)。与siErbB2组相比,siErbB2+IGF-1组ErbB2、Akt、PI3K、eNO S和Bcl-2的mRNA和蛋白表达量显著下降(P均<0.05),Caspase-3、Bax的mRNA和蛋白表达量显著增加(P均<0.05),细胞增殖能力、侵袭能力、迁移能力均明显下降(P均<0.05),细胞凋亡率上升(P<0.05)。结论ErbB2基因沉默可抑制PI3K/Akt/eNOS信号通路,从而抑制卵巢癌细胞增殖,促进细胞凋亡。而PI3K/Akt/eNOS信号通路激活可逆转ErbB2基因沉默作用,促进卵巢癌细胞增殖并抑制细胞凋亡。

Objective To investigate the effect of ErbB2 gene silencing on the biological characteristics of ovarian cancer cells by mediating the PI3K/Akt/eNOS signaling pathway and its mechanism.Methods Human ovarian cancer HO8910 cells in the logarithmic growth phase were divided into blank control group(without transfection of any sequence),negative control group(transfected with empty vector plasmid),siErbB2 group(transfected with siRNA sequence),wortmannin group(transfected with the PI3K/Akt/eNOS signaling pathway inhibitor wortmannin),and siErbB2+IGF-1 group(transfected with siRNA sequence and the PI3K/Akt/eNOS signaling pathway agonist IGF-1).Quantitative reverse transcription-polymerase chain reaction and Western blot were used to measure the mRNA and protein expression levels of ErbB2,Akt,PI3K,eNOS,Caspase-3,Bax,and Bcl-2,and the changes in cell proliferation,apoptosis,migration,and invasion were observed.Results Compared with the blank control group,the siErbB2 group and the wortmannin group had significant reductions in the mRNA and protein expression levels of Akt,PI3K,eNOS,and Bcl-2 and significant increases in the mRNA and protein expression levels of Caspase-3 and Bax in ova-rian cancer cells(F=3.58-8.69,P<0.05),as well as significant reductions in cell proliferation,invasion,and migration abilities(F=8.84-15.67,all P<0.05)and a significant increase in cell apoptotic rate(F=9.46,P<0.05);the siErbB2 group had a significant reduction in the expression of ErbB2(P<0.05).Compared with the siErbB2 group,the siErbB2+IGF-1 group had signi-ficant reductions in the mRNA and protein expression levels of ErbB2,Akt,PI3K,eNOS,and Bcl-2(all P<0.05),significant increases in the mRNA and protein expression of Caspase-3 and Bax(all P<0.05),significant reductions in cell proliferation,invasion,and migration abilities(all P<0.05),and a significant increase in cell apoptotic rate(P<0.05).Conclusion ErbB2 gene silencing can inhibit the PI3K/Akt/eNOS signaling pathway, thereby inhibiting the proliferation of ovarian cancer cells and promoting cell apoptosis. The activation of the PI3K/Akt/eNOS signaling pathway can reverse the effect of ErbB2 gene silencing, promote the proli-feration of ovarian cancer cells, and inhibit cell apoptosis.