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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 observe the changes of malignant phenotype of human SW480 colon cancer cells that inhibit the expression of DcR3 gene. METHODS: Small double-stranded DNA was constructed by RNA interference (RNAi) technology, cloned into expression vector, transfected into SW480 colon cancer cell line (DcR3 overexpressing cells) to form small interfering double-stranded RNA in cells. DcR3 mRNA was identified and degraded . The DcR3 low expression transfected cancer cells were screened, and the growth rate and apoptosis expression of DcR3-SW480-RNAi transfected cells in vitro were observed. Results: Transfection of DcR3-SW480-RNAi-F1R1 cells decreased the transcription of DcR3 mRNA. The product of gel electrophoresis showed that the bands of DcR3-SW480-RN-Ai-F1R1 cells were significantly decreased compared with the control group; Quantitative analysis of the cells showed that the number of DcR3-RNAi-SW480 colon cancer cells was significantly reduced, P <0.001; Western blot analysis showed that the expression of apoptotic antibodies Caspase-3 and PARP increased. Conclusion: DcR3 mRNA expression in transfected DcR3-RNAi-SW480 colon cancer cells is decreased. Tumor cells reduce the number of growth, increased expression of apoptosis, there is some exploration of anti-cancer prospects.