RNAi对结肠癌SW480细胞Nucleostemin基因表达的影响

目的:探求RNA技术干扰人结肠癌SW480细胞Nucleostemin(NS)基因后,结肠癌SW480细胞NucleosteminmRNA和蛋白水平的表达,及细胞增殖情况变化.方法:构建Nucleostemin特异性真核表达载体,和脂质体共同转染SW480细胞后,采用Real-timePCR和Westernblot方法检测NS基因mRNA和蛋白变化,MTT法检测细胞的增殖情况.结果:与对照组相比较,RNAi干扰细胞后NSmRNA表达明显下降;NS蛋白表达亦明显下降(1.069±0.368,1.003±0.313,1.901±0.817,P<0.05;1.069±0.368,1.003±0.313,2.035±0.665,P<0.05),空白对照组(仅加入脂质体)和阴性对照组(RNAi错义序列)间无显著差异(2.035±0.665,1.9001±0.817,P>0.05).MTT显示RNA干扰后细胞增殖明显受到抑制.结论:通过特异性RNA干扰NS基因后,结肠癌SW480细胞增殖受到抑制.

RNAi-mediated downregulation of expression of nucleostemin in human colorectal carcinoma cell line SW480

AIM:To investigate the effect of transfection of nucleostemin-specific small interfering RNA(siRNA)on nucleostemin expression and cell proliferation in human colorectal carcinoma cell line SW480.METHODS:A plasmid containing siRNA targeting the nucleostemin gene was constructed and transfected into SW480 cells with Lipofectamine2000 Reagent.After transfection,nucleostemin expression was detected by real-time PCR and Western blot,and cell proliferation was determined by MTT assay.RESULTS:Compared to the control groups,the levels of the nucleostemin mRNA and protein were obviously reduced in cells transfectedwith nucleostemin-specific RNA(protein:1.069 ±0.368,1.003±0.313 vs 1.901±0.817,both P< 0.05;mRNA:1.069±0.368,1.003±0.313 vs 2.035 ±0.665,both P<0.05).There were no significant differences in the expression of nucleostemin mRNA and protein between the two control groups(both P>0.05).SiRNA-mediated inhibition of nucleostemin expression significantly suppressed the proliferation of SW480 cells.CONCLUSION:SiRNA-mediated inhibition of nucleostemin expression significantly suppressed the proliferation of SW480 cells.