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STAT3小干扰RNA的构建及其对胃癌细胞增殖和凋亡的影响

, PP. 603-605

Keywords: STAT,RNA干扰,胃癌,增殖,凋亡

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Abstract:

目的以信号转导与转录激活因子3(signaltransducerandactivatoroftranscription3,STAT3)为靶基因,设计并构建短发夹状小干扰RNA表达载体,探讨STAT3小干扰RNA表达载体对胃癌细胞株SGC7901增殖和凋亡的影响。方法以PAVU6+27质粒为载体,构建STAT3的小干扰RNA表达载体,并将其转染胃癌SGC7901细胞,G418筛选后经反转录聚合酶链反应和Westernblot法检测STAT3基因和蛋白表达的改变,MTT法和3H-TdR掺入率检测细胞增殖和DNA合成情况,原位凋亡检测细胞凋亡率,流式细胞术检测抗细胞凋亡因子Bcl-2的表达。结果酶切鉴定和测序分析表明靶向STAT3的RNA干扰重组体构建成功;重组体转染SGC7901后,STAT3的mRNA和蛋白水平较空载体转染组显著下降;MTT比色A值和3H-TdR掺入率均明显低于对照组(P<0.05),凋亡率升高(P<0.01),Bcl-2表达下降。结论STAT3小干扰RNA表达载体可能是抑制人胃癌细胞增殖和促进凋亡的有效手段。

References

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