Objective: To construct retroviral vector with HBV Pol gene and study its expression in vitro. Methods:The recombinant plasmid HBV2/pBR322 containing 2 copies of HBV full-length gene was provided as the amplification ...Objective: To construct retroviral vector with HBV Pol gene and study its expression in vitro. Methods:The recombinant plasmid HBV2/pBR322 containing 2 copies of HBV full-length gene was provided as the amplification template. The PCR product was cloned into pMD 18-T and subcloned into pMSCVneo and pLNCX2 to construct the recombinant retroviral vectors with HBV Pol gene named by HBV P/pMSCVneo and HBV P/pLNCX2. HBV Pol gene was detected by RTPCR after transfecting the recombinant plasmids into SMMC7721 cells with liposome and G418 selection. Results: The retroviral vector with HBV Pol gene was successfully constructed and the expression of HBV Pol gene in vitro was detected by RTPCR. Conclusion: The retroviral vector with HBV Pol gene can be obtained, which will provide a new insight on the function of HBV Pol gene.展开更多
Alcohol consumption is a major health issue and associated with human cancers, such as liver and breast cancers. Alcohol was classed as carcinogen to human by IARC. We have performed in vivo and in vitro studies which...Alcohol consumption is a major health issue and associated with human cancers, such as liver and breast cancers. Alcohol was classed as carcinogen to human by IARC. We have performed in vivo and in vitro studies which demonstrate that diluted ethanol promotes cell proliferation and transformation and tumor formation. Consumption of liquor spirits (white wines) is a popular behavior. However, it is unclear whether liquor spirits affect cellular phenotypes of human cancers. At present study, we used diluted ethanol and liquor spirits (Sample #1 and Sample #2) to determine the changes in RNA polymerase III-dependent gene (Pol III gene) transcription, cell growth and colony formation in the different human cancer lines. The results indicate that low concentration of ethanol increases RNA Pol III gene transcription and rate of cell growth. However, both liquor spirits (Sample #1 and Sample #2) inhibit the activity of RNA Pol III genes and repress cell proliferation of the cancer lines, compared to diluted ethanol. The liquor spirits reduce the rate of colony formation of human breast cancer cells and esophageal carcinoma cells. The inhibitions of the liquor spirits to RNA Pol III genes, cell growth and colony formation are in a dose-dependent manner. These new findings suggest that the liquor spirits contain some active components to repress Pol III gene transcription and cell growth caused by ethanol in different human cancer cells.展开更多
基金Supported by National Natural Science Foundation of China (No. 30170856)
文摘Objective: To construct retroviral vector with HBV Pol gene and study its expression in vitro. Methods:The recombinant plasmid HBV2/pBR322 containing 2 copies of HBV full-length gene was provided as the amplification template. The PCR product was cloned into pMD 18-T and subcloned into pMSCVneo and pLNCX2 to construct the recombinant retroviral vectors with HBV Pol gene named by HBV P/pMSCVneo and HBV P/pLNCX2. HBV Pol gene was detected by RTPCR after transfecting the recombinant plasmids into SMMC7721 cells with liposome and G418 selection. Results: The retroviral vector with HBV Pol gene was successfully constructed and the expression of HBV Pol gene in vitro was detected by RTPCR. Conclusion: The retroviral vector with HBV Pol gene can be obtained, which will provide a new insight on the function of HBV Pol gene.
文摘Alcohol consumption is a major health issue and associated with human cancers, such as liver and breast cancers. Alcohol was classed as carcinogen to human by IARC. We have performed in vivo and in vitro studies which demonstrate that diluted ethanol promotes cell proliferation and transformation and tumor formation. Consumption of liquor spirits (white wines) is a popular behavior. However, it is unclear whether liquor spirits affect cellular phenotypes of human cancers. At present study, we used diluted ethanol and liquor spirits (Sample #1 and Sample #2) to determine the changes in RNA polymerase III-dependent gene (Pol III gene) transcription, cell growth and colony formation in the different human cancer lines. The results indicate that low concentration of ethanol increases RNA Pol III gene transcription and rate of cell growth. However, both liquor spirits (Sample #1 and Sample #2) inhibit the activity of RNA Pol III genes and repress cell proliferation of the cancer lines, compared to diluted ethanol. The liquor spirits reduce the rate of colony formation of human breast cancer cells and esophageal carcinoma cells. The inhibitions of the liquor spirits to RNA Pol III genes, cell growth and colony formation are in a dose-dependent manner. These new findings suggest that the liquor spirits contain some active components to repress Pol III gene transcription and cell growth caused by ethanol in different human cancer cells.
