论文部分内容阅读
目的: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:应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,同时分别设立人成纤维对照组、SW480对照组、SW480错配链-SECs组和SW480空转染试剂组。于48h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,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:SW480STAT3-SECs组的细胞可见凋亡小体,出现明显的凋亡现象,而人成纤维对照组、人成纤维STAT3-SECs组、SW480对照组、SW480错配链-SECs组和SW480空转染试剂组未出现明显的凋亡现象。SW480STAT3-SECs组细胞的凋亡比率较SW480对照组、SW480错配链-SECs组和SW480空转染试剂组有明显的增高。RT-PCR所得数据经统计学处理得出:SW480STAT3-SECs组细胞的STAT3基因表达在mRNA水平上显著低于SW480对照组(P<0.01);而人成纤维对照组与人成纤维STAT3-SECs组,SW480细胞对照组与SW480错配链-SECs组、SW480空转染试剂组之间无明显差异(P>0.05)。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: To study the effect of STAT3-siRNA on the apoptosis of colorectal cancer cells with STAT3 gene expression. METHODS: STAT3-siRNA expression cassettes (STAT3-siRNAs) were transfected into human colorectal cancer SW480 cells and human fibroblasts in vitro using lipofectamine transfection reagent. At the same time, human fibroblast control Group, SW480 control group, SW480 mismatch-SECs group and SW480 empty transfection reagent group. The cells were collected after 48h. The changes of cell surface were observed by fluorescence staining. The apoptosis of human colorectal cancer SW480 cells was detected by flow cytometry. Total RNA was extracted from the cells. The expression of STAT3 mRNA was detected by RT-PCR. expression. Results: Apoptotic bodies were observed in SW480STAT3-SECs group, and obvious apoptosis was observed. However, in human fibroblast control group, human fibroblast STAT3-SECs group, SW480 control group, SW480 mismatch-SECs group and SW480 null Transfection reagent group did not appear obvious apoptosis. The apoptosis rate of SW480STAT3-SECs group was significantly higher than that of SW480 control group, SW480 mismatch-SECs group and SW480 empty transfection reagent group. RT-PCR data obtained by statistical analysis: SW480STAT3-SECs cells STAT3 gene expression at the mRNA level was significantly lower than the SW480 control group (P <0.01); and human fibroblast control group and human fibroblast STAT3-SECs There was no significant difference (P> 0.05) between SW480 cell control group and SW480 mismatch-SECs group and SW480 empty transfection reagent group. Conclusion: The silencing of STAT3 gene by RNAi can reduce the expression of STAT3 in human colorectal cancer SW480 cells and induce cell apoptosis.