共同沉默DcR3和EGFR基因对结肠癌细胞株SW480体外生长的影响

目的 探讨共同沉默诱骗受体3(DcR3)和表皮生长因子(EGFR)基因对结肠癌细胞株SW480生长增殖的影响和诱导凋亡作用研究.方法 根据DcR3、EGFR的cDNA序列构建3组针对DcR3和EGFR的siRNA,分别使用脂质体Lipofectamine 2000转染的方法将各个siRNA导入结肠癌细胞株SW480中,48 h后通过实时荧光定量PCR检测各个siRNA对肿瘤靶基因mRNA表达的影响,采用Western-blot技术检测其靶基因蛋白的表达水平,筛选出对人结肠癌SW480细胞系中DcR3和EGFR抑制效果最强的siRNA,然后单独或联合转染SW480细胞,MTT法检测转染后对结肠癌细胞株SW480生长增殖活性的影响,流式细胞仪检测对结肠癌细胞株SW480的诱导凋亡作用.结果 共同沉默DcR3和EGFR基因对SW480细胞增殖的抑制作用优于DcR3或EGFR单基因沉默组(P<0.05).双基因共沉默组、DcR3单基因沉默组、EGFR单基因沉默组转染48 h后转染细胞的凋亡率分别为(20.1±1.7)％,(16.5±2.2)％及(15.9±1.3)％,差别具有统计学意义(P<0.05).结论 DcR3和EGFR双基因共沉默能有效抑制人结肠癌细胞株SW480的增殖并诱导其凋亡,且作用优于DcR3或EGFR单基因沉默.

The Effects of Co-silence of DcR3 and EGFR Genes on SW480 Cell Line Growth in vitro

Objective To explore the effects of co-silence of decoy receptor 3(DcR3)and epidermal growth factor receptor(EGFR)genes on the proliferation and apoptosis of SW480 cell line in vitro.Methods According to the siRNA design principle,three siRNA target sequences on DcR3 and EGFR genes were synthesized respectively.And siRNA was transfected into SW480 cells by Lipofectamine 2000 transfection methods.Real-time qPCR and Western-blot methods were used to detecte the mRNA and protein expression of DcR3 and EGFR of the SW480 cells after transfection for 48 hours.The highest inhibition efficiency siRNA sequences were selected and then transfected into SW480 cells using control-siRNA,DcR3-siRNA,EGFR-siRNA or DcR3-EGFR-siRNA co-silence.MTT method was used to measure cell proliferation and the flow-cytometry method was used to determine cell apoptosis rate after transfection.Results The highest inhibition rate on SW480 proliferation was co-silence of DcR3 and EGFR genes group compared with DcR3-siRNA or EGFR-siRNA group(P<0.05).And the DcR3-EGFR-siRNA co-silence group apoptosis rate(20.1±1.7)％were significantly increased compared with DcR3-siRNA group(16.5±2.2)％or EGFR-siRNA group(15.9±1.3)％(P<0.05).Conclusion Co-silence of DcR3 and EGFR genes can inhibit cell proliferation and induce cell apoptosis more effectively than DcR3 or EGFR genesilence singly.