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目的探讨抑制靶向转录信号传导子与激活子-3(STAT-3)基因表达对结肠癌细胞增殖以及侵袭转移行为的影响。方法实验分三组:空白对照组、转染NC-siRNA(阴性对照组)及转染特异性STAT-3-siRNA组(RNAi组)。人结肠癌细胞HCT116瞬时转染特异性siRNA,24、48、72 h后MTT法检测三组细胞的增殖活性,48 h后RQPCR检测STAT-3基因mRNA表达。RQ-PCR及细胞免疫化学法检测MMP-9在mRNA和蛋白水平的表达,Transwell小室法检测细胞体外侵袭能力。结果 RNAi组与其他两组相比细胞增殖受抑明显(均P<0.01);细胞生长减慢,RNAi组较阴性对照组STAT-3mRNA下降了71.91%(P<0.01)。RNAi组与其他两组相比MMP-9在mRNA和蛋白水平表达均明显下降(均P<0.01);侵袭实验RNAi组较空白对照组侵袭力下降了49.75%(P<0.01),而两对照组之间差异无统计学意义(P>0.05)。结论下调STAT-3基因表达可有效抑制结肠癌细胞的增殖和侵袭转移,抑制MMP-9在mRNA和蛋白水平的表达。沉默STAT-3基因抑制结肠癌细胞侵袭能力的机制之一可能是通过下调MMP-9途径。
Objective To investigate the effects of inhibiting the expression of target transcription signal transducer and activator of transcription-3 (STAT-3) gene on the proliferation, invasion and metastasis of colon cancer cells. Methods The experiment was divided into three groups: blank control group, transfected NC-siRNA (negative control group) and transfected specific STAT-3-siRNA group (RNAi group). Human colon cancer cell line HCT116 was transiently transfected with specific siRNA. The proliferative activity of the three groups of cells was detected by MTT assay at 24, 48 and 72 hours. The expression of STAT-3 mRNA was detected by RQPCR 48 hours later. RQ-PCR and immunocytochemistry were used to detect the expression of MMP-9 at mRNA and protein level. Transwell chamber assay was used to detect the invasion ability of cells in vitro. Results Compared with the other two groups, the cell proliferation was significantly inhibited in the RNAi group (all P <0.01). The cell growth slowed down, and the STAT-3 mRNA in the RNAi group decreased by 71.91% (P <0.01) compared with the negative control group. Compared with the other two groups, the mRNA and protein levels of MMP-9 in RNAi group were significantly decreased (all P <0.01), and invasiveness of RNAi group was decreased by 49.75% (P <0.01) compared with the control group There was no significant difference between groups (P> 0.05). Conclusion Down-regulation of STAT-3 gene expression can effectively inhibit the proliferation, invasion and metastasis of colon cancer cells and inhibit the expression of MMP-9 at mRNA and protein levels. One of the mechanisms by which silencing the STAT-3 gene inhibits the invasiveness of colon cancer cells may be through downregulation of the MMP-9 pathway.