摘要
目的:应用RNA干扰技术抑制COX-2基因,观察其对肝癌细胞HepG2生长的影响.方法:构建靶向抑制COX-2基因表达的短发夹双链RNA(shRNA)真核表达载pshRNA-COX-2,转染肝癌细胞HepG2,用RT-PCR和Western Blot分别从mRNA和蛋白水平检测其对COX-2的抑制效应,MTT法检测对HepG2细胞生长的影响.结果:与未转染组相比,pshR-NA-COX-2重组表达质粒抑制了HepG2细胞COX-2mRNA及蛋白表达,抑制率分别为69.9%和50.3%;并可抑制HepG2细胞生长,72h后有统计学差异(P<0.05).结论:pshRNA-COX-2重组表达载体能显著抑制肝癌细胞COX-2基因表达,从而抑制肿瘤细胞增殖.
AIM： To investigate the effects of inhibition of cyclooxygenase-2 （COX-2） with RNA interference （RNAi） on the growth of hepatocellular carcinoma HepG2 cells. METHODS： The eukaryotic expression plasmid of short hairpin RNA （shRNA） targeting COX-2 gene （pshRNA-COX-2） was constructed and transfected into hepatocellular carcinoma HepG2 cells; the expressions of COX-2 mRNA and protein were detected with reverse transcriptional polymerase chain reaction （RT-PCR） and Western Blot assay, respectively. The growth of HepG2 cells was detected by MTY. RESULTS： Compared with untransfected group, recombinant expression vector pshRNA-COX-2 resulted in the reduction of COX-2 mRNA and protein by 69.9% and 50.3% respectively, and the growth of HepG2 cells was significantly inhibited after 72 h （ P 〈 0.05 ） . CONCLUSION ： Recombinant expression vector pshRNA-COX-2 can significantly inhibit the expression of COX-2 gene , and suppress the growth of tumor cells.
出处
《第四军医大学学报》
北大核心
2007年第14期1272-1274,共3页
Journal of the Fourth Military Medical University
基金
重庆市自然科学基金项目(2006BB5271)
关键词
RNA干扰
重组表达载体
环氧化酶-2
肝肿瘤
RNA interference
recombinant expression vcectors
cyclooxygenase-2
liver neoplasms
