摘要
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.
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 tar- geting the nucleostemin gene was constructed and transfected into SW480 cells with Lipo- fectamine2000 Reagent. After transfection, nu- cleostemin 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 inhibi- tion of nucleostemin expression significantly suppressed the proliferation of SW480 cells. CONCLUSION： SiRNA-mediated inhibition of nucleostemin expression significantly sup- pressed the proliferation of SW480 cells.
出处
《世界华人消化杂志》
CAS
北大核心
2012年第1期70-73,共4页
World Chinese Journal of Digestology
