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(−)-Epigallocatechin-3-gallate inhibits human papillomavirus (HPV)-16 oncoprotein-induced angiogenesis in non-small cell lung cancer cells by targeting HIF-1α

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Abstract

Purpose

To investigate the effects of (−)-epigallocatechin-3-gallate (EGCG) on human papillomavirus (HPV)-16 oncoprotein-induced angiogenesis in non-small cell lung cancer (NSCLC) cells and the underlying mechanisms.

Methods

NSCLC cells (A549 and NCI-H460) transfected with EGFP plasmids containing HPV-16 E6 or E7 oncogene were treated with different concentrations of EGCG for 16 h. The effects of EGCG on angiogenesis in vitro and in vivo were observed. The expression of HIF-1α, p-Akt, and p-ERK1/2 proteins in NSCLC cells was analyzed by Western blot. The levels of HIF-1α mRNA in NSCLC cells were detected by real-time RT-PCR. The concentration of VEGF and IL-8 in the conditioned media was determined by ELISA. HIF-1α, VEGF, and CD31 expression in A549 xenografted tumors of nude mice was analyzed by immunohistochemistry.

Results

HPV-16 E6 and E7 oncoproteins HIF-1α-dependently promoted angiogenesis in vitro and in vivo, which was inhibited by EGCG. Mechanistically, EGCG inhibited HPV-16 oncoprotein-induced HIF-1α protein expression but had no effect on HIF-1α mRNA expression in NSCLC cells. Additionally, 50 and 100 μmol/L of EGCG significantly reduced the secretion of VEGF and IL-8 proteins induced by HPV-16 E7 oncoprotein in NSCLC A549 cells. Meanwhile, HPV-16 E6 and E7 oncoproteins HIF-1α-dependently enhanced Akt activation in A549 cells, which was suppressed by EGCG. Furthermore, EGCG inhibited HPV-16 oncoprotein-induced HIF-1α and HIF-1α-dependent VEGF and CD31 expression in A549 xenografted tumors.

Conclusions

EGCG inhibited HPV-16 oncoprotein-induced angiogenesis conferred by NSCLC through the inhibition of HIF-1α protein expression and HIF-1α-dependent expression of VEGF, IL-8, and CD31 as well as activation of Akt, suggesting that HIF-1α may be a potential target of EGCG against HPV-related NSCLC angiogenesis.

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Acknowledgments

This work was supported by the grants from National Natural Science Foundation of China, 81073103 and 30872944 (To X. Tang), Guangdong Natural Science Foundation, S2012010008232 (To X. Tang), Science and Technology of Guangdong Province, 2009B030801330 (To X. Tang), the Specialized Foundation for Introduced Talents of Guangdong Province Higher Education, 2050205 (To X. Tang), Zhanjiang Municipal Governmental Specific Financial Fund Allocated for Competitive Scientific and Technological Projects, 2012C0303-56 (To X. Tang), and Science and Technology Innovation Fund of Guangdong Medical College, STIF201105 (To K. Zhou).

Conflict of interest

None declared.

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Authors and Affiliations

  1. Institute of Biochemistry and Molecular Biology, Guangdong Medical College, 2 Wenming Donglu, Xiashan, Zhanjiang, 524023, Guangdong, People’s Republic of China

    Li He, Erying Zhang, Jingli Shi, Xiangyong Li, Keyuan Zhou, Qunzhou Zhang & Xudong Tang

  2. Guangdong Key Laboratory of Medical Molecular Diagnosis, Guangdong Medical College, Zhanjiang, 524023, People’s Republic of China

    Xiangyong Li, Keyuan Zhou & Xudong Tang

  3. Department of Oral Surgery and Pharmacology, University of Pennsylvania School of Dental Medicine, Philadelphia, PA, 19104, USA

    Qunzhou Zhang & Anh D. Le

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  1. Li He
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Correspondence to Xudong Tang.

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Li He and Erying Zhang contributed equally to this work.

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He, L., Zhang, E., Shi, J. et al. (−)-Epigallocatechin-3-gallate inhibits human papillomavirus (HPV)-16 oncoprotein-induced angiogenesis in non-small cell lung cancer cells by targeting HIF-1α. Cancer Chemother Pharmacol 71, 713–725 (2013). https://doi.org/10.1007/s00280-012-2063-z

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