含人_干扰素基因真核表达载体的构建及其在肝癌细胞中的表达_郭英.pdfVIP

11 1 Vol.11 No. 1 2002 1 Chinese Journal of General Surgery Jan. 2002 : 1005- 6947(2002) 01- 0026- 03 - 1 1 2 2 2 郭英 , 杨丽新 , 王坤 , 闵军, 陈积圣 ( 1. 中山医科大学附属第三医院外科, 广东 广州510630; 2. 孙逸仙纪 医院外科, 广东 广州510120) : - ( IFN- ) ( pcDNA3- IFN- ) , IFN- cDNA ( pcDNA3) , pcDNA3- IFN- SMMC- 772 1 QGY- 7701 , G4 18 R - PCR IFN- , ELISA IFN - IFN - mRNA pcDNA3- IFN- IFN- IFN- : / ; ; , ; , : R7 35. 7; R978. 7 : A Expression of exogenous interferon- gene in human hepatocellular carci- noma cells mediated by liposome 1 2 2 2 GUO Ying , YANG Li-xin, WANG kun , MIN Jun , CHEN Ji-sheng ( 1. Dep artment of Surgery , The Third Aff iliated Hosp ital , Sun Yat - sen University of Medical Science, Guang hou 510630, China ; 2. Dep ar tment of Surgery , Sun Yat - sen Memorial Hosp ital , Guang hou 510120, China) Abstract:Objective o establish a recombinant eukaryotic expression vector containing human interferon - ( IFN - ) cDNA , and invest igate the expression of IFN - gene in transfected hepatocellular carcinoma ( HCC) cell lines . Methods pcDNA3 - IFN - , established by subcloning IFN - cDNA into a mam- malian expression vector pcDNA3 , was mediated into HCC cell lines SMMC - 7 72 1 and QGY - 7 70 1 by lipofectamine. he transfected HCC cells w ere selected in RPMI 1640 containing G418( 4 00 ~ 70 0g ) . R - PCR analysis and ELISA assay w ere used for mRNA transcription and protein expression of IFN - gene. Results IFN - mRNA w as detected only in pcDNA3 - IFN - transfected cell lines , and high levels of IFN protein were det ected only in the supernatants of IFN- gene- modified cell lines. Conclusions pcD- NA3- IFN- can stably express IFN- cDNA in the tra