DcR3-RNAi对人SW-480结肠癌细胞系恶性表型的影响

Effect of malignant phenomenon on humen SW480 colonic cancer cell line by DcR3-RNA interference

目的:观察研究抑制DcR3基因表达的人SW480结肠癌细胞其恶性表型的改变。方法:应用RNA干扰(RNAi)技术,构建小双链DNA,克隆入表达载体,转染进入SW480结肠癌细胞系(DcR3高表达细胞),在细胞内形成小干扰双链RNA;识别并降解DcR3mRNA。筛选DcR3低表达转染癌细胞,3H观测其体外DcR3-SW480-RNAi转染细胞的增长率及凋亡表达。结果:转染的DcR3-SW480-RNAi-F1R1细胞可降低DcR3mRNA的转录,凝胶电泳反应产物显示与对照组相比,DcR3-SW480-RN-Ai-F1R1细胞组条带明显减弱;3H掺入细胞的定量分析显示DcR3-RNAi-SW480结肠癌细胞的数量明显减少,P<0.001;Western印记检测显示凋亡抗体Caspase-3及PARP表达增加。结论:经转染的DcR3-RNAi-SW480结肠癌细胞其DcR3mRNA表达降低。肿瘤细胞的生长数量减少,凋亡表达增加,有一定可探索性抗癌前景。

OBJECTIVE： To study the relationship between the depressed DcR3 gene expression and effect of malignant pheno type on SW480 colonic carcinima cell line. METHODS： A small in terfering double strand DcR3 RNA was constructed by using the RNAi method, and then cloned into vector ＂pSilencer 2.1 Hygro＂, and transfected into SW480 colonic cancer cells which expressed a high level of DcR3. The small double strand RNA was recognized and reduced as DcR3 mRNA. The DcR3 low-expression transfec tion cancer cells were examined on their growth rate. RESULTS： The transcription of DcR3-mRNA was decreased by DcR3-RNAi-SW480 transfection cancer cells, and agarose gel electrophoresis of the reaction products showed there was a weaker band with DcR3-RNAi F1R1 cells compared with the control groups, a H TdR incorporation analysis showed the growth rate of DcR3-RNAi-F1R1 tumor cells also reduced （P〈0. 001）. The expression of apoptotic antibody Caspase-3 and PARP were increased by Western blotting. CONCLUSION ： That DeR3-RNAi SW480 trasfection colonic carcinoma cells with transcription of DcR3 mRNA are de creased may have some explored anticancer prospect.