siRNA沉默STAT3基因对肝癌细胞HepG2增殖的影响

摘要

目的 通过体外沉默信号传导及转录活化因子3(signal transducer and activator of transcription 3,STAT3)基因转录,探讨阻断JAK2/STAT3信号通路对肝癌细胞HepG2增殖能力的影响及其机制.方法 siRNA体外沉默肝癌细胞株HepG2 STAT3基因转录,检测细胞内STAT3 mRNA和蛋白的表达变化证实基因沉默效果,MTT检测细胞增殖能力的变化,流式细胞仪检测细胞周期,RT-PCR检测JAK2/STAT3信号通路下游分子survivin及caspase3表达的变化.结果 阻断JAK2/STAT3信号通路后肝癌细胞株HepG2增殖能力下降,细胞分裂停滞于G0-G1期,伴随survivin转录下降,caspase3转录上升.结论 阻断JAK2/STAT3信号通路可抑制肝癌细胞增殖,可能同调控survivin及caspase3有关.%Objective To study the proliferation change of HepG2 cells by inhibiting the expression of signal transducer and activator of transcription 3 (STAT3) in vitro and to investigate its underlying molecular mechanism. Methods siRNA against STAT3 inhibited the expression and activation of STAT3 protein in HepG2 cells in vitro; cell proliferation and cycle were detected by MTT assay and flow cytometry respectively; transcriptions of survivin and caspase3 were detected by reverse transcription PCR. Results Proliferation of HepG2 cells was inhibited by blocking JAK2/STAT3 signaling pathway; cell division was arrested at G0-G1 phase. Transcription of survivin was downregulated while that of caspase3 was upregulated. Conclusion Proliferation and division of HepG2 cells can be inhibited by blocking JAK2/STAT3 signaling pathway, the mechanism of which may be related to be regulation of survivin and caspase3.