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Blocking activity of the HPV18 virus in cervical cancer cells using the CRISPR/Cas9 system

Jing Wang1, Meng Guo2, Quanxing Wang2

  • 1Changzheng Hospital, Second Military Medical University Shanghai, China.

International Journal of Clinical and Experimental Pathology
|January 18, 2020
PubMed
Abstract

Insights

CRISPR/Cas9 technology effectively silenced HPV18 E6 and E7 oncogenes in HeLa cells. This gene editing approach halted cell cycle progression and reduced cancer cell proliferation, offering a potential therapeutic strategy for cervical cancer.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Gene Editing

Background:

  • Human Papillomavirus type 18 (HPV18) oncogenes E6 and E7 are critical drivers of cervical cancer development.
  • The CRISPR/Cas9 system offers a precise tool for gene editing and functional studies.

Purpose of the Study:

  • To design and validate specific single-guide RNA (sgRNA) sequences targeting HPV18 E6 and E7 oncogenes using the CRISPR/Cas9 system.
  • To investigate the impact of E6 and E7 oncogene knockout on HeLa cervical cancer cell proliferation and cell cycle progression.

Main Methods:

  • Construction and transfection of lentivirus vectors encoding Cas9 and sgRNAs targeting HPV18 E6 and E7 into HeLa cells.
  • Quantification of E6 and E7 mRNA and protein expression using RT-PCR and Western blot.
  • Analysis of cell cycle distribution via flow cytometry and evaluation of cell proliferation using colony formation assays.

Main Results:

  • Successfully constructed and transfected sgRNA vectors targeting E6 and E7 oncogenes, leading to significant inhibition of their mRNA and protein expression.
  • E6-sgRNA2 and E7-sgRNA1 demonstrated the highest inhibition rates (85% and 86% for mRNA, respectively).
  • CRISPR/Cas9-mediated E6 and E7 knockdown resulted in increased G1/G0 phase cell population and significantly reduced HeLa cell colony formation.

Conclusions:

  • The CRISPR/Cas9 system effectively targets and silences HPV18 E6 and E7 oncogenes in cervical cancer cells.
  • Inhibition of E6 and E7 expression leads to cell cycle arrest and decreased proliferation in HeLa cells.
  • This study validates CRISPR/Cas9 as a potent tool for targeting oncogenes and provides insights into cervical cancer therapeutic strategies.
Keywords:
Cervical cancerE6E7clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9)human papillomavirus 18

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