Treatment with Radix Euphorbiae Ebracteolatae Significantly Decreases the Expression of E6 and L1, and Increases the Expression of p53 and Rb in HPV18-infected Human Foreskin Keratinocytes

Author(s): Xiang He, Xufeng He, Ping Xu, Lili Yang, Xin Ma, Wen Li, Huimin Zhang*

Journal Name: Current Molecular Medicine

Volume 19 , Issue 1 , 2019

  Journal Home
Translate in Chinese
Become EABM
Become Reviewer
Call for Editor


Background: Radix Euphorbiae Ebracteolatae (REE) was recently reported to be significantly superior to vitamin A acid ointment in treating multiple plantar warts. However, the effects of REE on HPV18 remain unclear. Therefore, the current study aimed to investigate the effects of REE on the proliferation of HPV18, and explore possible molecular mechanisms underlying the effects.

Methods: HFK and HFK-HPV18 were treated with water-extracted single or compound REE, ethanol-extracted single or compound REE, TNF-α and IFN for 3 days, respectively. In addition, the organotypic rafts containing HFK-HPV18 and HFK were treated with REE, IFN and TNF-α for 7 days, respectively. Cell proliferation rates were measured with Brdu. mRNA expression of E6, L1, p53 and Rb was detected by qPCR. Protein expression of p53, Rb and L1 was detected by Western blot.

Results: Compared to HFK group, HFK-HPV18 group had significantly higher expression of E6 and L1. Compared to the control group, HFK-HPV18 treated with REE, TNF-α and IFN displayed significantly lower proliferation rates. The mRNA expression of E6 was markedly lower, and mRNA expression of p53 and Rb was significantly higher after treatment of REE in HFK-HPV18 or in organotypic rafts containing HFK-HPV18. Treatment with REE markedly increased the protein expression of p53 and Rb, and decreased the protein expression of L1 in HFK-HPV18 or in organotypic rafts containing HFK-HPV18. Among all formula of REE, the inhibition of proliferation rates and expression of E6 and L1, and the increase in expression of p53 and Rb in HFK-HPV18 was highest in ethanol-extracted compound REE group.

Conclusions: The proliferation rates are significantly lower in HFK-HPV18 treated with REE. The expression of E6 and L1 is markedly lower, and expression of p53 and Rb is significantly higher after REE treatment in HFK-HPV18 or organotypic rafts containing HFK-HPV18. Among all formula of REE, ethanol-extracted compound REE displays the highest protection against HPV18.

Keywords: Radix Euphorbiae Ebracteolatae, HPV18, cancer, condyloma acuminata, Keratinocytes, HFK.

Ljubojevic S, Skerlev M. HPV-associated diseases. Clin Dermatol 2014; 32: 227-34.
Sawaya GF, Kulasingam S, Denberg TD, Qaseem A. Cervical Cancer Screening in Average-Risk Women: Best Practice Advice From the Clinical Guidelines Committee of the American College of Physicians. Ann Intern Med 2015; 162: 851-9.
Zhao K, Xu G, Jin R, Xu L. [Taxonomic studies on original plants of radix Euphorbiae ebracteolatae]. Zhongguo Zhongyao Zazhi 1995; 20: 5-6, 61.
He X, Xu P, He XF, et al. Efficacy of topical administration of Radix Euphorbiae Ebracteolatae on multiple plantar warts: A parallel randomized trial. Chin J Integr Med 2017.
Hoppe-Seyler K, Bossler F, Braun JA, Herrmann AL, Hoppe-Seyler F. The HPV E6/E7 Oncogenes: Key Factors for Viral Carcinogenesis and Therapeutic Targets. Trends Microbiol 2017; 26: 158-68.
Ershov VA, Il’inskaia EV. [Capsid protein L1 HPV-16 in a modified cervical epithelium]. Vopr Onkol 2013; 59: 47-51.
Buck CB, Day PM, Trus BL. The papillomavirus major capsid protein L1. Virology 2013; 445: 169-74.
Duerksen-Hughes PJ, Yang J, Schwartz SB. HPV 16 E6 blocks TNF-mediated apoptosis in mouse fibroblast LM cells. Virology 1999; 264: 55-65.
Thyrell L, Sangfelt O, Zhivotovsky B, et al. The HPV-16 E7 oncogene sensitizes malignant cells to IFN-alpha-induced apoptosis. J Interferon Cytokine Res 2005; 25: 63-72.
Satsuka A, Yoshida S, Kajitani N, Nakamura H, Sakai H. Novel human papillomavirus type 18 replicon and its application in screening the antiviral effects of cytokines. Cancer Sci 2010; 101: 536-42.
Wang S, Long S, Xiao S, Wu W, Hann SS. Decoction of Chinese Herbal Medicine Fuzheng Kang-Ai Induces Lung Cancer Cell Apoptosis via STAT3/Bcl-2/Caspase-3 Pathway. Evid Based Complement Alternat Med 2018; 2018: 8567905.
Gong X, Zhang Y, Pan J, Qu H. Optimization of the ethanol recycling reflux extraction process for saponins using a design space approach. PLoS One 2014; 9: e114300.
Motobu M, El-Abasy M, Na KJ, Hirota Y. Detection of mitogen-induced lymphocyte proliferation by bromodeoxyuridine (BrdU) incorporation in the chicken. J Vet Med Sci 2002; 64: 377-9.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method Methods 2001; 25: 402-8.
Jeong DE, Lee Y, Lee SV. Western Blot Analysis of C. elegans Proteins. Methods Mol Biol 2018; 1742: 213-25.
Wilson R, Laimins LA. Differentiation of HPV-containing cells using organotypic “raft” culture or methylcellulose. Methods Mol Med 2005; 119: 157-69.
Bruno MT, Ferrara M, Fava V, Rapisarda A, Coco A. HPV genotype determination and E6/E7 mRNA detection for management of HPV positive women. Virol J 2018; 15: 52.
Ram BM, Dolpady J, Kulkarni R, et al. Human papillomavirus (HPV) oncoprotein E6 facilitates Calcineurin-Nuclear factor for activated T cells 2 (NFAT2) signaling to promote cellular proliferation in cervical cell carcinoma. Exp Cell Res 2017; 362: 132-41.
Thatte J, Massimi P, Thomas M, Boon SS, Banks L. The HPV E6 PDZ Binding Motif links DNA Damage Response Signaling to E6 Inhibition of p53 Transcriptional Activity. J Virol 2018; 92(16): e00465-18.
Kojima Y, Otsuki N, Kubo M, et al. Adenovirus-mediated transfer of HPV 16 E6/E7 antisense RNA combined with cisplatin inhibits cellular growth and induces apoptosis in HPV-positive head and neck cancer cells. Cancer Gene Ther 2018; 25(9-10): 274-83.
Norman I, Hjerpe A, Andersson S. High-risk HPV L1 capsid protein as a marker of cervical intraepithelial neoplasia in high-risk HPV-positive women with minor cytological abnormalities. Oncol Rep 2013; 30: 695-700.
Xu HM, Sun WY, Zhang G, Ni XH. [Detection and significance of HPV L1 capsid protein in cervical squamous intraepithelial lesions]. Zhonghua Bing Li Xue Za Zhi 2011; 40: 549-50.
Lee HJ, Yoon JK, Heo Y, et al. Therapeutic potential of an AcHERV-HPV L1 DNA vaccine. J Microbiol 2015; 53: 415-20.
Munagala R, Kausar H, Munjal C, Gupta RC. Withaferin A induces p53-dependent apoptosis by repression of HPV oncogenes and upregulation of tumor suppressor proteins in human cervical cancer cells. Carcinogenesis 2011; 32: 1697-705.
Guo KY, Han L, Li X, et al. Novel proteasome inhibitor delanzomib sensitizes cervical cancer cells to doxorubicin-induced apoptosis via stabilizing tumor suppressor proteins in the p53 pathway. Oncotarget 2018; 8: 114123-35.
Yang X, Lu L. Expression of HPV-16 E6 protein and p53 inactivation increases the uterine cervical cancer invasion. Drug Res (Stuttg) 2014; 65: 70-3.
Das S, Somasundaram K. Therapeutic potential of an adenovirus expressing p73 beta, a p53 homologue, against human papilloma virus positive cervical cancer in vitro and in vivo. Cancer Biol Ther 2006; 5: 210-7.
Sharma G, Rana NK, Singh P, Dubey P, Pandey DS, Koch B. p53 dependent apoptosis and cell cycle delay induced by heteroleptic complexes in human cervical cancer cells. Biomed Pharmacother 2017; 88: 218-31.
Sima N, Wang W, Kong D, et al. RNA interference against HPV16 E7 oncogene leads to viral E6 and E7 suppression in cervical cancer cells and apoptosis via upregulation of Rb and p53. Apoptosis 2008; 13: 273-81.
Deshpande R, Mansara P, Kaul-Ghanekar R. Alpha-linolenic acid regulates Cox2/VEGF/MAP kinase pathway and decreases the expression of HPV oncoproteins E6/E7 through restoration of p53 and Rb expression in human cervical cancer cell lines. Tumour Biol 2015; 37: 3295-305.
Zingue S, Nde CBM, Michel T, et al. Ethanol-extracted Cameroonian propolis exerts estrogenic effects and alleviates hot flushes in ovariectomized Wistar rats. BMC Complement Altern Med 2017; 17: 65.
Xie XL, Gi M, Fujioka M, et al. Ethanol-extracted propolis enhances BBN-initiated urinary bladder carcinogenesis via non-mutagenic mechanisms in rats. Food Chem Toxicol 2015; 83: 193-200.
Doi K, Fujioka M, Sokuza Y, et al. Chemopreventive Action by Ethanol-extracted Brazilian Green Propolis on Post-initiation Phase of Inflammation-associated Rat Colon Tumorigenesis. In Vivo 2017; 31: 187-97.

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2019
Published on: 25 February, 2019
Page: [20 - 31]
Pages: 12
DOI: 10.2174/1566524019666190226102713
Price: $65

Article Metrics

PDF: 27