Journal of Shandong University (Health Sciences) ›› 2018, Vol. 56 ›› Issue (12): 1-6.doi: 10.6040/j.issn.1671-7554.0.2018.369

   

Construction of stable-expressed cell lines of human papillomavirus early oncoprotein E6/E7 and its effect on the expression of Rab12

ZHANG Wenhao1,2, QIAO Lijun2, ZHENG Jingyi1, Jason Chen2, ZHANG Weifang1   

  1. 1. Department of Pathogenic Biology;
    2. Cancer Research Centre, Cheeloo College of Medical, Shandong University, Jinan 250012, Shandong, China
  • Published:2022-09-27

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Abstract: Objective To explore the impact of early oncoprotein E6/E7, encoded by human papillomavirus(HPV), on the expression of Rab12 protein, a small GTP enzyme protein. Methods The high titer of virus particles, containing pBabe-Vector plasmid and pBabe-16E7 plasmid, were obtained with retroviruses packing system. UMSCC 10A cell line was infected with the virus particles and selected by puromycin to get UMSCC 10A-16E7cell line, which stably expressed HPV16E7. Western blotting was used to detect the expression of E7 associated protein, including p53 and E2F1, to verify the infection. The expression level of Rab12 in UMSCC 10A-Vector and UMSCC 10A-16E7 cell lines was compared. Further verification was conducted in HaCaT and RPE1 cell lines as above. The expression of 16E6/E7 in SiHa cell line and Caski cell line(HPV-16 positive cervical cancer cell line), and the expression of 18E6/E7 in Hela cell line(HPV-18 positive cervical cancer cell line)were interfered by siRNA. Western blotting was used to compare the expression level of E6 and E7 associated protein(E2F1, p53 and Rab12)after the interference. Results These stable-expressed cell lines were successfully built up with retroviruses packaging system. The expression of Rab12 increased in E6/E7 over-expression cells, and decreased in the E6/E7 interfere cells. Conclusion Early oncoprotein E6/E7 coded by HPV can regulate the expression of Rab12.

Key words: Rab12, Human papillomavirus, E6, E7, Retroviruses packaging system

CLC Number: 

  • R737
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