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RNA Interference of Human Papillomavirus Type 16 E7 Increases HLA Class I Antigen Expression in HaCaT-E7 Cells
  1. Xiao-Mei Deng, PhD*,
  2. Wei Li, PhD*,
  3. Xiao Zhang, PhD,
  4. Chuan-Xin Wang, MD, PhD*,
  5. Zhao-Gang Dong, PhD*,
  6. Xin Zhang, PhD*,
  7. Gui-Xi Zheng, PhD*,
  8. Xu-Hua Zhang, PhD,
  9. Ni Zheng, MASc*,
  10. Li-Li Wang, PhD*,
  11. Lu-Tao Du, MASc* and
  12. Shun Wang, MASc*
  1. * Department of Clinical Laboratory, Qilu Hospital, Shandong University, Jinan, China;
  2. Weifang Center for Disease Control and Prevention, Weifang, China; and
  3. Department of Clinical Laboratory, The Second Hospital of Shandong University, Jinan, China.
  1. Address correspondence and reprint requests to Chuan-Xin Wang, MD, PhD, Department of Clinical Laboratory, Qilu Hospital, Shandong University, 107 Wenhua Rd, Jinan 250012, China. E-mail: cxwang{at}sdu.edu.cn.

Abstract

Background: High-risk human papillomaviruses (HPVs) are the major causative agents of cervical cancer. The E7 protein of high-risk HPV disturbs cell cycle control and down-regulates components of the antigen presentation pathway, suggesting an ideal target for development of the immunotherapy in HPV-positive cervical cancers. We previously reported that HPV16 E7 could down-regulate cell-surface HLA class I antigen accompanying decreased expression of transporter associated with antigen processing 1 (TAP-1). The purpose of this study was to determine whether knockdown of HPV16 E7 could up-regulate surface HLA class I antigen expression in HPV16 E7 expressing HaCaT cells (HaCaT-E7).

Methods: An E7-specific small interfering RNA (siRNA) was transfected into the HaCaT-E7 cells, and the expression of HPV16 E7 was measured by real-time reverse transcriptase polymerase chain reaction and Western blot. With the use of flow cytometry analysis, the levels of cell surface HLA class I antigen and intracellular TAP-1 expression were detected.

Results: It was found that transfection of HPV16 E7-siRNA reduced HPV16 E7 expression as measured on messenger RNA and protein levels. The flow cytometry analysis showed that, compared with mock transfection, a statistically significant increase of approximately 75% in surface HLA class I levels was observed in HaCaT-E7 cells at 72 hours after transfection of E7 siRNA. Moreover, he knockdown of E7 in HaCaT-E7 cells could result in an increase of intracellular TAP-1 expression, which is essential for the expression of HLA class I at cell surface.

Conclusions: Our study showed that the knockdown of HPV16 E7 could increase cell surface HLA class I antigen expression in HaCaT-E7 cells. In addition, for HPV-positive human cervical cancer, our observations indicate that the HPV E7 gene is a target of choice.

  • HPV16 E7
  • RNA interference
  • HLA class I
  • TAP
  • Cervical cancer
  • HaCaT cell

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