靶向EZH2的shRNA表达对SW480细胞的抑制作用及5-Fu对其影响的研究

Inhibitive effect of EZH2-specific shRNA on SW480 cells and the impact of 5-FU

目的研究EZH2特异性shRNA对人结肠癌细胞株SW480细胞的抑制作用和化疗药物5-FU对其干预作用的影响。方法体外将EZH2特异性shRNA表达载体转染SW480细胞,RT-PCR和Western blotting检测细胞EZH2 mRNA及蛋白的表达,M TT法检测细胞增殖活性;将靶向EZH2的shRNA表达的SW480细胞接种于裸鼠,5-FU对其进行干预,RT-PCR检测瘤组织EZH2 mRNA的表达。结果与阴性对照组相比,基因沉默组EZH2 mRNA表达明显降低(P<0.01),EZH2蛋白表达明显降低(P<0.05);与空白对照组相比,基因沉默组细胞生长受到明显抑制(P<0.05);接种于裸鼠后,与正常组相比,5-FU基因沉默组与基因沉默组瘤组织体积、质量、EZH2 mRNA的表达均明显降低(P<0.01),与基因沉默组相比,5-FU基因沉默组EZH2 mRNA的表达明显降低(P<0.05)。结论靶向EZH2的shRNA表达在体外及体内均可显著抑制SW480细胞EZH2 mRNA和蛋白表达及细胞生长;5-FU能显著抑制体内EZH2基因沉默的SW480细胞EZH2 mRNA的表达。

Objective To investigate the inhibitive effect of EZH2-specific shRNA on human colon cancer cell line SW480 and the impact of 5-FU on this inhibitive effect.Methods SW480 cells were transfected with EZH2-specific shRNA in vitro,then EZH2 mRNA and protein expressions were detected with RT-PCR and Western blotting,and cell proliferation was examined with MTT.The EZH2-specific shRNA expressing SW480 cells were implanted to nude mice and treated with 5-FU.After that,the levels of EZH2 mRNA expressed by tumor tissues were measured with RT-PCR. Results Compared with that of the negative control group,the EZH2 mRNA expression （P〈0.01 ）and protein expression （P〈0.05）of the gene-silencing group markedly decreased.The cell growth of the gene-silencing group was inhibited compared with that of the control group （P〈0.05 ）.In nude mice,the tumor volume,tumor weight and EZH2 mRNA expression of the 5-FU gene-silencing group and of the gene-silencing group remarkably decreased com-pared with those of the normal group （P〈0.01）.The EZH2 mRNA expression of the 5-FU gene-silencing group signifi-cantly decreased compared with that of the gene-silencing group （P〈0.05）.Conclusion The EZH2-specific shRNA＆nbsp;plays a significant inhibitive role on EZH2 mRNA and protein expressions in SW480 cells both in vitro and in vivo. 5-FU can significantly inhibit the EZH2 mRNA expression of EZH2-gene-silencing SW480 cells in vivo.