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.
出处
《中国生物制品学杂志》
CAS
CSCD
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)
