Bcl-2、C-Raf及双靶点RNAi质粒对HCT-8细胞株增殖的抑制作用  

Inhibitory Effect of Bcl-2,C-Raf and Double Target RNAi Plasmid on Proliferation of HCT-8 Cell Strain

作　　者：王艳芬[1] 迟春萍[2] 王志武[2] 时成波[2] 吉海滨[2] 牛灵[2] 盛军[2] 李东复[1] 

机构地区：[1]吉林大学第二临床医院消化科,长春130041 [2]长春生物制品研究所,长春130062

出　　处：《中国生物制品学杂志》2007年第1期47-50,共4页Chinese Journal of Biologicals

摘　　要：目的研究Bcl-2、C-Raf及双靶点的RNA干扰质粒对人结肠癌HCT-8细胞株增殖的抑制作用。方法利用Ambion公司网上设计软件,分别以Bcl-2和C-Raf为靶标,设计合成63个核苷酸的双链RNA,构建质粒pSilencer3·1-H1/Bcl-2、pSilencer3·1-H1/C-Raf及pSilencer3·1-H1/Bcl-2/C-Raf,分别转染结肠癌细胞株HCT-8,用MTT法检测其对细胞增殖的抑制作用,RT-PCR检测Bcl-2和C-Raf的mRNA转录。结果经MTT法检测,构建的Bcl-2和C-Raf基因及双靶点RNAi质粒3个转染组的细胞存活率均降低,双靶点质粒转染组较两个单靶点转染组细胞存活率明显降低,差异有显著意义。经RT-PCR检测,转染双靶点质粒后,HCT-8细胞中Bcl-2和C-Raf基因的表达均较对照组明显减弱。结论Bcl-2、C-Raf及双靶点RNAi质粒对HCT-8细胞株增殖有抑制作用。Objective To study the inhibitory effect of Bcl-2, C-Raf and double target RNAi plasmid on the proliferation of human colorectal carcinoma HCT-8 cells. Methods Design and synthesize a double-stranded RNA consisting of 63 nucleotides using Bcl- 2 and C-Raf as target separately and construct plasmids pSilencer 3. 1-H1/Bcl-2 ,pSilencer 3. 1-H1/C-Raf and pSilencer 3.1-H1/Bcl- 2/C-Raf. Transfect HCT-8 cells with the three constructed plasmids respectively and determine the inhibitory effect on proliferation of cells by MTT. Test the transcription of Bcl-2 and C-Raf mRNA by RT-PCR. Results All the survival rates of HCT-8 cells transfected with the three constructed plasmids decreased. However, the survival rate of HCT-8 cells transfected with pSilencer 3. 1-H1/Bcl-2/C- Raf was significantly lower than those with pSilencer 3. 1-H1/Bcl-2 and pSilencer 3. 1-H1/C-Raf. RT-PCR proved that the expression levels of Bcl-2 and C-Raf genes in HCT-8 cells transfected with pSilencer 3. 1-H1/Bcl-2/C-Raf decreased signiticanfly as compared with blank control. Conclusion Bcl-2,C-Raf and double target RNAi plasmid inhibited the proliferation of HCT-8 cells.

关 键 词：BCL-2基因 C-Raf基因 RNA干扰 基因沉默 

分 类 号：Q786[生物学—分子生物学] R735.35[医药卫生—肿瘤]