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【目的】应用经小干扰RNA(short interfere RNA,siRNA)表达盒介导的RNAi对结直肠癌细胞中的STAT3基因进行干涉作用以研究STAT3-siRNA对STAT3基因表达阳性的结直肠癌细胞凋亡的影响。【方法】用脂质体转染试剂将STAT3-siRNA表达盒(STAT3-siRNA expression cassettes,STAT3-SECs)体外转染至人结直肠癌SW480细胞及人成纤维细胞中。于48 h后收集细胞,先经荧光染色方法观察细胞表象变化,再通过流式细胞仪检测人结直肠癌SW480细胞凋亡情况,最后分别提取细胞总RNA,用RT-PCR测定STAT3基因在mRNA水平的表达。【结果】SW480 STAT3-SECs组的细胞可见凋亡小体,出现明显的凋亡现象,而其它实验组均未出现明显的凋亡现象。流式细胞术结果显示SW480 STAT3-SECs组中凋亡细胞百分比由0.2%增加至26.0%;S期细胞比率由32.3%下降至9.2%。在mRNA水平上,SW480 STAT3-SECs组细胞的STAT3基因表达减少了(62.16±12.50)%,显著低于SW480对照组(P<0.01)。【结论】应用RNAi技术沉默STAT3基因可以降低人结直肠癌SW480细胞中STAT3的表达,诱导细胞的凋亡。
【Objective】 The purpose of this study was to investigate the interference of STAT3 gene in colorectal cancer cells by RNAi mediated by short interference RNA (siRNA) expression cassette to study the apoptosis of colorectal cancer cells with STAT3 gene expression by STAT3-siRNA Impact. 【Method】 STAT3-siRNA expression cassettes (STAT3-SECs) were transfected into human colorectal cancer SW480 cells and human fibroblasts in vitro using lipofectamine transfection reagent. The cells were collected after 48 h. The changes of cell appearance were observed by fluorescence staining. The apoptosis of human colorectal cancer SW480 cells was detected by flow cytometry. Finally, the total RNA was extracted and the expression of STAT3 mRNA was detected by RT- Horizontal expression. 【Results】 Apoptotic bodies were observed in SW480 STAT3-SECs group, with obvious apoptosis, but no obvious apoptosis in other experimental groups. The results of flow cytometry showed that the percentage of apoptotic cells in SW480 STAT3-SECs group increased from 0.2% to 26.0%, and the percentage of S phase cells decreased from 32.3% to 9.2%. At the mRNA level, the STAT3 gene expression in SW480 STAT3-SECs group decreased by (62.16 ± 12.50)%, which was significantly lower than that in SW480 control group (P <0.01). 【Conclusion】 Silencing STAT3 gene by RNAi can reduce the expression of STAT3 in human colorectal cancer SW480 cells and induce cell apoptosis.