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[Establishment and identification of human hepatocellular carcinoma line stably expressing hepatitis C virus core
Qinghua Qiao1, Xin Lv2, Yingfeng Lei2
1Department of Microbiology, School of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032; Department of Clinical Laboratory, Chinese PLA 152 Hospital, Pingdingshan 467000, China.
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.
Conclusion:
The SMMC-7721 human hepatocellular carcinoma cell line stably expressing HCV core protein has been established successfully.
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