[Establishment and identification of human hepatocellular carcinoma line stably expressing hepatitis C virus core protein].
OBJECTIVE
To establish SMMC-7721 human hepatocellular carcinoma cell line stably expressing hepatitis C virus (HCV) core protein.


METHODS
A lentiviral vector containing HCV core gene was constructed and transfected into HEK293T cells to package recombinant lentivirus (rLV-core) containing ZsGreen and HCV core genes. The SMMC-7721 cells were infected with the rLV-core. The expression of HCV core mRNA was examined by real-time fluorescent quantitative PCR and the HCV core protein was detected by immunofluorescence cytochemistry and Western blotting. The stably transfected cell line was screened.


RESULTS
The lentiviral vector was confirmed by enzyme digestion and sequencing. The green fluorescence was seen under fluorescence microscope 48 hours after virus packaging. The SMMC-7721 cell line stably expressing HCV core protein was obtained after infected with the rLV-core. Real-time PCR showed the expression of HCV core mRNA, and both immunofluorescence cytochemistry and Western blotting verified the expression of HCV core protein.


CONCLUSIONS
The SMMC-7721 human hepatocellular carcinoma cell line stably expressing HCV core protein has been established successfully.
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032; Department of Clinical Laboratory, Chinese PLA 152 Hospital, Pingdingshan 467000, China.
Qinghua
Q
Qiao
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Xin
X
Lv
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Yingfeng
Y
Lei
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Jing
J
Yang
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Min
M
Yao
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Peijun
P
Han
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Pinxiang
P
Dang
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Haiwei
H
Zhao
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Daihui
D
Weng
Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Wen
W
Yin
D006528:Carcinoma, Hepatocellular / Q000821:virology*; D045744:Cell Line, Tumor; D005455:Fluorescent Antibody Technique; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008113:Liver Neoplasms / Q000821:virology*; D016133:Polymerase Chain Reaction; D014162:Transfection; D014758:Viral Core Proteins / Q000032:analysis / Q000235:genetics*
D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus