Construction of human recombinant lentiviral vector of GP73and its expression in DC2.4 cells  

作　　者：Xi-Lei Li Man-Ya Wu Xiao-Yun Shen Ye-Bin Pang Yan Feng Bi-Yu Cui Xiao-Ling Luo 

机构地区：[1]Department of Experimental Research,The Affiliated Tumor Hospital of Guangxi Medical University [2]Department of Radiotherapy,The affiliated Zhongshan Hospital of Fudan University

出　　处：《广西医科大学学报》2016年第1期1-5,共5页Journal of Guangxi Medical University

基　　金：supported by the National Natural Science Foundation of China (No.81360347);the Key University and College Project of Guangxi Provisional Department of Education(No.ZD2014027).

摘　　要：Objective:This study aimed to prepare Golgi protein-73(GP73)-DC vaccine by constructing human lentiviral vector of GP73and transfecting this vector in vitro into a murine dendritic cell line DC 2.4.This will lay the foundation for further study on targeted therapy of hepatocellular carcinoma using GP73-DC vaccine.Methods:Human GP73 gene was cloned into lentivirus GV358vector and validated by polymerase chain reaction(PCR)and sequencing.The validated GV358-GP73vector was then cotransfected with pHelper 1.0plasmid into 293Tcells,in order to prepare lentivirus GP73vector.Finally,the protein expression of GP73 was achieved by transfecting the lentivirus GP73vector into DC 2.4cells in vitro and determined by Western blotting.Results:PCR and sequencing confirmed that the GV358-GP73lentivirus vector was constructed correctly,and the titer level reached 2×108TU/mL.After transfection in DC 2.4cells,the GP73protein level was significantly enhanced.Conclusion:The lentivirus GV538-GP73vector was constructed successfully,and could be effectively expressed in DC 2.4cells.Objective：This study aimed to prepare Golgi protein-73（GP73）-DC vaccine by constructing human lentiviral vector of GP73and transfecting this vector in vitro into a murine dendritic cell line DC 2.4.This will lay the foundation for further study on targeted therapy of hepatocellular carcinoma using GP73-DC vaccine.Methods：Human GP73 gene was cloned into lentivirus GV358vector and validated by polymerase chain reaction（PCR）and sequencing.The validated GV358-GP73vector was then cotransfected with pHelper 1.0plasmid into 293Tcells,in order to prepare lentivirus GP73vector.Finally,the protein expression of GP73 was achieved by transfecting the lentivirus GP73vector into DC 2.4cells in vitro and determined by Western blotting.Results：PCR and sequencing confirmed that the GV358-GP73lentivirus vector was constructed correctly,and the titer level reached 2×108TU/mL.After transfection in DC 2.4cells,the GP73protein level was significantly enhanced.Conclusion：The lentivirus GV538-GP73vector was constructed successfully,and could be effectively expressed in DC 2.4cells.

关 键 词：食管癌 根治术 切口 比较 

分 类 号：R730.3[医药卫生—肿瘤]