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目的探讨信号转导子和转灵激活子3(STAT3)小干扰RNA(siRNA)对食管鳞状细胞癌细胞株(EC9706和Eca109)中持续性激活STAT3信号的阻断情况及阻断STAT3信号对细胞增殖的影响。方法将化学合成的100 nmol/L的STAT3 siRNA转染EC9706和Eca109细胞,逆转录聚合酶链反应检测转染前后STAT3 mRNA的表达,Western blot检测转染前后STAT3及磷酸化STAT3 (p-STAT3)蛋白的表达,凝胶电泳迁移率检测转染前后活化STAT3蛋白的核结合情况,力甲基偶氮唑蓝(MTT)检测转染前后细胞增殖能力的改变,流式细胞仪检测转染前后细胞周期的改变。结果STAT3 siRNA以时间依赖方式特异性地抑制STAT3 mRNA及STAT3、p-STAT3蛋白的表达,并且STAT3蛋白的核结合活性下降,使细胞周期阻滞于G_0/G_1期,细胞增殖受到明显的抑制。与对照组相比EC9706细胞转染后72 h G_0/G_1期的细胞增加了16.1%,同时S期细胞减少11.1%;Eca109细胞转染后72 h G_0/G_1期的细胞增加了11.8%,同时S期细胞也较对照组明显减少。结论STAT3 siRNA能够特异地阻断食管鳞癌细胞中STAT3信号的持续性激活并抑制肿瘤细胞的增殖。
OBJECTIVE: To investigate the effects of signal transducer and activator of transcription 3 (STAT3) interfering RNA on the persistence of STAT3 signaling in esophageal squamous cell carcinoma cell lines (EC9706 and Eca109) Effect of cell proliferation. Methods 100 nmol / L STAT3 siRNA was transfected into EC9706 and Eca109 cells. The expression of STAT3 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) before and after transfection. STAT3 and p-STAT3 (phospho-STAT3) Protein expression and electrophoretic mobility shift assay were used to detect the nuclear binding of activated STAT3 protein before and after transfection. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the proliferation of cells before and after transfection. Flow cytometry Cycle changes. Results STAT3 siRNA specifically inhibited STAT3 mRNA and STAT3, p-STAT3 protein expression in a time-dependent manner, and STAT3 protein nuclear binding activity decreased, cell cycle arrest in G_0 / G_1 phase, cell proliferation was significantly inhibited. Compared with the control group, the number of cells in G0 / G1 phase at 72 h after transfection of EC9706 cells increased by 16.1% and the cells in S phase decreased by 11.1%. The cells in G0 / G1 phase at 72 h after transfection of Eca109 cells increased by 11.8% S phase cells were also significantly reduced compared with the control group. Conclusion STAT3 siRNA can specifically block the continuous activation of STAT3 signal in esophageal squamous cell carcinoma and inhibit the proliferation of tumor cells.