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The impact of HCV core protein on the expression of lncRNA and circRNA in human Huh7 hepatoma cell line and the preliminary mechanism study

Jiefu Peng  Di Wu  Miao Li  Songyan Wu Wu  Xiaoyu Yu  Fei Li  Jipei Liao  Jun Dou  
【摘要】:Background: Hepatitis C Virus(HCV) has a closely correlation with chronic hepatitis, liver cirrhosis and hepatocellular carcinoma(HCC). As one of the most important fectors, HCV core protein constructs the capsid of HCV, and it participates in immune escape of HCV, resisting to the drug therapy and the liver diseases like live fibrosis. Objective: To investigate the impact of HCV core protein to the expression of lncRNA and the relevant preliminary carcinogenic mechanism.Methods: Use the core protein of 1b HCV to transfect into the Huh7 cells, by selection of G418, stable transfection cells will be harvested and RT-PCR will be performed to detect the expression of core potein. Then the core stable expression cells and contrast cells will be send to the company to analyze by the lncRNA and circRNA microarray, molecules which up-regulate or down-regulate more than 2 fold will be chosen, then bioinformatic analysis will be performed to find the possible correlation between these molecules and the changes of physiology and pathology. Results: The data showed that 4851 lncRNAs were up-regulated and 3569 lncRNAs were down-regulated more than two folds compared to the control genes; 4785 mRNAs were up-regulated and 3192 mRNAs were down-regulated more than two folds compared with the control genes. 823 and 419 circRNAs were up-regulated or down-regulated respectively. In addtion, the KEGG pathway analysis indicated that these genes have some connection with the endoplasmic reticulum stress, apoptosis, calcium singling pathway, Wnt singling pathway, HBV and HCV infection pathway as well as other pathways. Moreover, miR122, miR192, miR204 expression vectors were constructed and transfected into the Huh7-core cell lines successfully, and their regulation to the target genes were verified by qPCR. Conclusion:Mir192-HOTTIP-GLS, Circ0001498-Mir122-MAP3K2, these two possible pathways may be related to the carcinogenesis of HCV core protein.

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