Silencing of HPV 18 oncoproteins with RNA interference causes growth inhibition of cervical cancer cells

Jayanthi S. Lea, Noriaki Sunaga, Mitsuo Sato, Geetha Kalahasti, David S. Miller, John D. Minna, Carolyn Y. Muller

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Silencing the expression of human papillomavirus (HPV) oncoproteins should have therapeutic benefits for cervical cancer. The authors' objective was to study RNA interference of the HPV 18 E6/E7 bicistronic mRNA with E6 small interfering RNA (siRNA) and E7 siRNA and determine the effect of each siRNA on oncoprotein expression, resultant cell growth, and downstream molecular effects. RNA interference was used to knockdown HPV 18 E6 and E7 oncoproteins on the HPV 18 positive cervical cancer cell lines HeLa and C4I. Western blotting was used to assay for each oncoprotein expression and select downstream molecular targets. Cell cycle analyses, cell viability assays, and colony formation assays were performed to determine the effect of treatment by both HPV 18 E6 siRNA and E7 siRNA. The transfection reagent oligofectamine and Tax siRNA were used as negative controls. Transfection with E6 siRNA caused complete loss of E6 but not E7 oncoprotein. However, E7 siRNA induced complete loss of both E6 and E7 oncoproteins. E6 siRNA mediated the reexpression of p53 protein and a moderate decrease in phosphorylated retinoblastoma protein expression (pRb), resulting in decreased colony formation. Transfection with E7 siRNA mediated a robust increase in p53 expression and complete loss of pRb, resulting in a marked decrease in colony formation compared to the E6 siRNA (P =.001). Flow cytometry revealed significantly increased apoptotic cells with E7 siRNA compared to E6 siRNA and control. RNA interference targeting the E7 portion of the bicistronic HPV 18 mRNA can silence both E6 and E7 oncoproteins and is most effective in cervical cancer growth inhibition.

Original languageEnglish (US)
Pages (from-to)20-28
Number of pages9
JournalReproductive Sciences
Volume14
Issue number1
DOIs
StatePublished - Jan 2007

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Human papillomavirus 18
Oncogene Proteins
RNA Interference
Uterine Cervical Neoplasms
Small Interfering RNA
Growth
Transfection
Retinoblastoma Protein
Messenger RNA
Cell Survival

Keywords

  • Cervical cancer
  • HPV
  • RNA interference
  • Targeted therapy

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Silencing of HPV 18 oncoproteins with RNA interference causes growth inhibition of cervical cancer cells. / Lea, Jayanthi S.; Sunaga, Noriaki; Sato, Mitsuo; Kalahasti, Geetha; Miller, David S.; Minna, John D.; Muller, Carolyn Y.

In: Reproductive Sciences, Vol. 14, No. 1, 01.2007, p. 20-28.

Research output: Contribution to journalArticle

Lea, Jayanthi S. ; Sunaga, Noriaki ; Sato, Mitsuo ; Kalahasti, Geetha ; Miller, David S. ; Minna, John D. ; Muller, Carolyn Y. / Silencing of HPV 18 oncoproteins with RNA interference causes growth inhibition of cervical cancer cells. In: Reproductive Sciences. 2007 ; Vol. 14, No. 1. pp. 20-28.
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