RNA干扰BC047440基因表达对HCT-8细胞增殖能力的影响  

Effect of Expression of RNA Interference Gene BC047440 on Proliferation Ability of HCT-8 Cells

作　　者：唐亮[1,2] 赵权[1] 宫鹏涛[2] 李建华[2] 杨举[2] 张西臣[2] 

机构地区：[1]吉林农业大学动物科技学院,长春130118 [2]吉林大学畜牧兽医学院,长春130062

出　　处：《中国生物制品学杂志》2010年第11期1207-1209,1216,共4页Chinese Journal of Biologicals

基　　金：国家重大基础研究计划资助项目(2006CB910505)

摘　　要：目的探讨RNA干扰结肠癌相关基因BC047440的表达对结肠癌细胞HCT-8增殖能力的影响。方法将BC047440基因shRNA干扰质粒pGPU6/GFP/Neo-BC047440-331(a1)、pGPU6/GFP/Neo-BC047440-451(a2)、pGPU6/GFP/Neo-BC047440-615(a3)、pGPU6/GFP/Neo-BC047440-756(a4)采用脂质体法转染结肠癌细胞HCT-8,荧光显微镜观察转染效率,荧光定量PCR法检测细胞BC047440基因mRNA的表达水平,MTT法分析细胞的增殖活性。结果细胞的转染效率为53%;干扰质粒a1、a2、a3、a4转染的HCT-8细胞BC047440基因mRNA的表达水平分别下降了36.1%、47.0%、53.8%和63.3%,a4质粒的干扰效率最高;细胞的增殖能力也明显下降。结论 BC047440基因RNA干扰质粒可明显抑制HCT-8细胞BC047440基因mRNA的表达及细胞的增殖,可能成为抑制结肠癌细胞生长的新基因。Objective To investigate the effect of colon cancer-related RNA interference gene BC047440 on the proliferation ability of colon cancer cells.Methods The shRNA interference plasmids pGPU6/GFP/Neo-BC047440-331（a1）,pGPU6/GFP/Neo-BC047440-451（a2）,pGPU6/GFP/Neo-BC047440-615（a3）and pGPU6/GFP/Neo-BC047440-756（a4）were transfected to colon cancer HCT-8 cells in mediation of liposome,then observed for transfection efficacy by fluorescent microscopy,determined for the expression level of BC047440 mRNA by fluorescent quantitative PCR,and analyzed for cell proliferation activity by MTT method.Results The transfection efficacy of HCT-8 cells was 53%.The expression levels of BC047440 mRNA in the HCT-8 cells transfected with RNA interference plasmids a1,a2,a3 and a4 decreased by 36.1%,47.0%,53.8% and 63.3% respectively,indicating the highest interference efficacy of a4 in the four plasmids.The proliferation ability of HCT-8 cells also decreased significantly.Conclusion The RNA interference plasmid carrying BC047440 gene inhibited the BC047440 mRNA expression as well as proliferation of HCT-8 cells,which might be used as a novel gene inhibiting the growth of colon cancer cells.

关 键 词：BC047440基因 结肠肿瘤 RNA干扰 

分 类 号：R735.35[医药卫生—肿瘤] Q786[医药卫生—临床医学]