摘要
目的 探讨HCV核心蛋白对肝瘤细胞P16、P2 7表达的影响。方法 将HCV核心基因cDNA插入真核表达载体PBK CMV的HindⅢ和BamHⅠ位点间 ,构建重组质粒PBK HCVc。再将重组质粒PBK HCVc和空载体分别导入肝癌细胞株HepG2中 ,G418筛选 ,RT PCR、蛋白印迹鉴定HCV核心蛋白表达。免疫组化 ,蛋白印迹检测P16、P2 7蛋白表达 ;RT PCR检测P16、P2 7mRNA。结果 重组质粒PBK HCVc在HepG2细胞中有稳定表达 ;表达HCV核心蛋白的细胞HepG2 C的P16、P2 7在蛋白水平和mRNA水平较转染空载体的细胞HepG2 CMV明显下降。结论 HCV核心蛋白能抑制P16、P2 7表达 。
Objective To explore the effect of HCV core protein on P16 and P27 expressions in HepG2 cells. Methods The HCV core gene cDNA was recoverd by PCR, and cloned into PBK CMV. The recombinant plasmid(PBK HCVc) and the empty vector were transfected into HepG2 cells with liposome. Screening was performed with G418. HCV core protein expression was detected by reverse transcription PCR and Western blotting. The protein expressions of P16 and P27 were analyzed by immunohistochemistry and Western blotting. P16 and P27mRNA expressions were analyzed by reverse transcription PCR. Results Stable expression of the recombinant plasmid was found in HCV core protein. The P16 and P27 of HepG2 C cells were lower than those of HepG2 CMV. Conclusion HCV core protein can inhibit the expressions of P16 and P27 in HepG2 cells, suggesting that HCV core protein contributes to viral carcinogenesis.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2003年第2期135-137,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 3990 0 176 )
关键词
丙型肝炎病毒
病毒核心蛋白
肝细胞癌
端粒酶
hepatitis C virus
viral core protein
hepatocellular carcinoma
P16
P27
