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Original Research

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HPV16 infection up-regulates Piwil2, which affects cell proliferation and invasion in cervical cancer by regulating MMP-9 via the MAPK pathway

  • W. Ling1,*,
  • H. Zhigang2
  • H. Tian3
  • Z. Bin3
  • X. Xiaolin3
  • Z. Hongxiu3

1Department of Gynaecology and Obstetrics, General Hospital of Lanzhou Military Region, Lanzhou, China

2Division of Medical Administration, General Hospital of Lanzhou Military Region, Lanzhou, China

3Department of Gynaecology and Obstetrics, Southwest Hospital of the Third Military Medical University, Chongqing, China

DOI: 10.12892/ejgo2666.2015 Vol.36,Issue 6,December 2015 pp.647-654

Published: 10 December 2015

*Corresponding Author(s): W. Ling E-mail: linowang@163.com

Abstract

Purpose of investigation: The present study aimed to investigate the effect of Piwil2 on proliferation and invasion of cervical cancer cells. Materials and Methods: Thirty-two HPV-positive or negative cervical cancer tissues and corresponding normal adjacent cervical tissues were obtained from General Hospital of Lanzhou Military Region. Piwil2 expression in these tissue samples, as well as two cervical cell lines were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemical. A specific short hairpin RNA (shRNA) was used to knockdown the Piwil2 gene in SiHa cells. CCK-8 assay and flow cytometry (FCM) was used to evaluate cell proliferation. Cell invasion was detected by transwell chambers assays. Immunoblotting was used to assess the effect on relevant proteins. Result: In the early stage (I A1 – I B1) of curvival, 84.4% (27/32) tumor tissues have a more predominant expression of Piwil2 than the normal adjacent samples. Piwil2 overexpression was correlated with HPV16 infection (p < 0.05). Knockdown of Piwil2 gene in SiHa cells inhibited cell growth and invasion, and downregulated matrix metalloproteinase- 9 (MMP-9) compared to scrambled shRNA transfected cells. Further analysis revealed that downregulation of Piwil2 gene induced inhibition of the MAPK signaling pathway activity. Conclusion: Piwil2, which stimulated by HPV16 infection, plays an important role in regulating proliferation and invasion of cervical cells by regulating MMP-9 expression via alternation of the MAPK signaling pathway.

Keywords

Cervical cancer; Piwil2; Cell growth; Invasion; MMP-9.

Cite and Share

W. Ling,H. Zhigang,H. Tian,Z. Bin,X. Xiaolin,Z. Hongxiu. HPV16 infection up-regulates Piwil2, which affects cell proliferation and invasion in cervical cancer by regulating MMP-9 via the MAPK pathway. European Journal of Gynaecological Oncology. 2015. 36(6);647-654.

References

[1] Siegel R., DeSantis C., Virgo K., Stein K., Mariotto A., Smith T., et al., “Cancer treatment and survivorship statistics, 2012”. CA Cancer J. Clin., 2012, 62, 220.

[2] Ferlay J., Shin H.R., Bray F., Forman D., Mathers C., Parkin D.M.: “Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008”. Int. J. Cancer, 2010, 127, 2893.

[3] Scatchard K., Forrest J.L., Flubacher M., Cornes P., Williams C.: “Chemotherapy for metastatic and recurrent cervical cancer”. Cochrane Database Syst. Rev., 2012, 10, CD006469.

[4] Bosch, F.X., M.M. Manos, N. Munoz, M. Sherman, A.M. Jansen, J. Peto, et al., “Prevalence of human papillomavirus in cervical cancer: a worldwide perspective. International biological study on cervical cancer (IBSCC) Study Group”. J. Natl., Cancer Inst., 1995, 87, 796.

[5] de Vet H.C., van Leeuwen F.E.: “Dietary guidelines for cancer pre-vention: the etiology of a confused debate”. Nutr. Cancer, 1986, 8, 223.

[6] Tindle R.W.: “Immune evasion in human papillomavirus-associated cervical cancer”. Nat. Rev. Cancer, 2002, 2, 59.

[7] Syrjanen K.J.: “Spontaneous evolution of intraepithelial lesions ac-cording to the grade and type of the implicated human papillomavirus (HPV)”. Eur. J. Obstet. Gynecol. Reprod. Biol., 1996, 65, 45.

[8] Burger R.A., Monk B.J., Kurosaki T., Anton-Culver H., Vasilev S.A., Berman M.L., Wilczynski S.P.: “Human papillomavirus type 18: as-sociation with poor prognosis in early stage cervical cancer”. J. Natl. Cancer Inst., 1996, 88, 1361.

[9] Soderlund-Strand A., Kjellberg L., Dillner J.: “Human papillo-mavirus type-specific persistence and recurrence after treatment for cervical dysplasia”. J. Med. Virol., 2014, 447, 300.

[10] Inaba K., Nagasaka K., Kawana K., Arimoto T., Matsumoto Y., Tsu-ruga T., et al.: “High-risk human papillomavirus correlates with re-currence after laser ablation for treatment of patients with cervical intraepithelial neoplasia 3: A long-term follow-up retrospective study”. J. Obstet. Gynaecol. Res., 2014, 40, 554.

[11] Sasaki T., Shiohama A., Minoshima S., Shimizu N.: “Identification of eight members of the Argonaute family in the human genome small star, filled”. Genomics, 2003, 82, 323.

[12] Kuramochi-Miyagawa S,. Kimura T., Ijiri T.W., Isobe T., Asada N., Fujita Y., et al.: “Mili, a mammalian member of piwi family gene, is essential for spermatogenesis”. Development, 2004, 131, 839.

[13] Liu X., Sun Y., Guo J., Ma H., Li J., Dong B., et al.: “Expression of hiwi gene in human gastric cancer was associated with proliferation of cancer cells”. Int. J. Cancer, 2006, 118, 1922.

[14] He G., Chen L., Ye Y., Xiao Y., Hua K., Jarjoura D., et al.: “Piwil2 expressed in various stages of cervical neoplasia is a potential complementary marker for p16”. Am. J. Transl. Res., 2010, 2, 156.

[15] Liu J.J., Shen R., Chen L., Ye Y., He G., Hua K., et al.: “Piwil2 is ex-pressed in various stages of breast cancers and has the potential to be used as a novel biomarker”. Int. J. Clin. Exp. Pathol., 2010, 3, 328.

[16] Yin D.T., Li H.Q., Wang Y.F., Cao S.L., Zhou Y.B., Zheng L.Y., et al.: “Expression of Piwil2 and its relationship with tumor invasion and metastasis in papillary thyroid carcinoma”. Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi, 2011, 46, 237.

[17] Li D., Sun X., Yan D., Huang J., Luo Q., Tang H., Peng Z.: “Piwil2 modulates the proliferation and metastasis of colon cancer via regulation of matrix metallopeptidase 9 transcriptional activity”. Exp. Biol. Med. (Maywood), 2012, 237, 1231.

[18] Lee J.H., Jung C., Javadian-Elyaderani P., Schweyer S., Schutte D., Shoukier M., et al.: “Pathways of proliferation and antiapoptosis driven in breast cancer stem cells by stem cell protein piwil2”. Cancer Res., 2010, 70, 4569.

[19] Oh S.J., Kim S.M., Kim Y.O., Chang H.K.: “Clinicopathologic Im-plications of PIWIL2 Expression in Colorectal Cancer”. Korean J. Pathol., 2012, 46, 318.

[20] Greither T., Koser F., Kappler M., Bache M., Lautenschlager C., Gobel S., et al.: “Expression of human Piwi-like genes is associated with prognosis for soft tissue sarcoma patients”. BMC Cancer, 2012, 12, 272.

[21] Feng D., Peng C., Li C., Zhou Y., Li M., Ling B., et al.: “Identification and characterization of cancer stem-like cells from primary carcinoma of the cervix uteri”. Oncol. Rep., 2009, 22, 1129.

[22] Choo K.B., Cheung W.F., Liew L.N., Lee H.H., Han S.H.: “Presence of catenated human papillomavirus type 16 episomes in a cervical carcinoma cell line”. J. Virol., 1989, 63, 782.

[23] Gierke P., Zhao C., Brackmann M., Linke B., Heinemann U., Braunewell K.H.: “Expression analysis of members of the neuronal calcium sensor protein family: combining bioinformatics and West-ern blot analysis”. Biochem. Biophys. Res. Commun., 2004, 323, 38.

[24] Maddirela D.R., Kesanakurti D. , Gujrati M., Rao J.S.: “MMP-2 sup-pression abrogates irradiation-induced microtubule formation in endothelial cells by inhibiting alphavbeta3-mediated SDF-1/CXCR4 signaling”. Int. J. Oncol., 2013, 42, 1279.

[25] Bizari, L., Borim A.A., Leite K.R., Goncalves Fde T., Cury P.M., Ta-jara E.H., Silva A.E.: “Alterations of the CCND1 and HER-2/neu (ERBB2) proteins in esophageal and gastric cancers”. Cancer Genet. Cytogenet., 2006, 165, 41.

[26] Lee J.H., Schutte D., Wulf G., Fuzesi L., Radzun H.J., Schweyer S., et al.: “Stem-cell protein Piwil2 is widely expressed in tumors and inhibits apoptosis through activation of Stat3/Bcl-XL pathway”. Hum. Mol. Genet., 2006, 15, 201.

[27] Baussano I., Elfstrom K.M., Lazzarato F., Gillio-Tos A., De Marco L., Carozzi F., et al.: “Type-specific human papillomavirus biological features: validated model-based estimates”. PLoS One, 2013, 29, 81171.

[28] de Sanjose S., Serrano B., Castellsague X., Brotons M., Muñoz J., Bruni L., Bosch F.X.: “Human papillomavirus (HPV) and related cancers in the Global Alliance for Vaccines and Immunization (GAVI) countries. A WHO/ICO HPV Information Centre Report”. Vaccine, 2012, 30, D1.

[29] Lorenz L.D., Rivera Cardona J., Lambert P.F.: “Inactivation of TP53 rescues the maintenance of high risk HPV DNA genomes deficient in expression of E6”. PLoS Pathog., 2013, 9, e1003717.

[30] Geiger T., Levitzki A.: “Loss of robustness and addiction to IGF1 during early keratinocyte transformation by human papilloma virus 16”. PLoS One, 200,m 2, e605.

[31] Lu Y., Zhang K., Li C., Yao Y., Tao D., Liu Y., et al.: “Piwil2 sup-presses TP53 by inducing phosphorylation of signal transducer and activator of transcription 3 in tumor cells”. PLoS One, 2012, 7, e30999.

[32] Tarin D., Matsumura Y.: “Recent advances in the study of tumour invasion and metastasis”. J. Clin. Pathol., 1994, 47, 385.

[33] Pilcher B.K., Dumin J.A., Sudbeck B.D., Krane S.M., Welgus H.G., Parks W.C.: “The activity of collagenase-1 is required for keratinocyte migration on a type I collagen matrix”. J. Cell. Biol., 1997, 137, 1445.

[34] Westermarck J., Kahari V.M.: “Regulation of matrix metallopro-teinase expression in tumor invasion”. FASEB J., 1999, 13, 781.

[35] Coussens L.M., Tinkle C.L., Hanahan D., Werb Z.: “MMP-9 sup-plied by bone marrow-derived cells contributes to skin carcinogenesis”. Cell, 2000, 103, 481.

[36] Bergers G., Brekken R., McMahon G., Vu T.H., Itoh T., Tamaki K., et al.: “Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis”. Nat. Cell. Biol., 2000, 2, 737-44.

[37] Nuovo G.J., MacConnell P.B., Simsir A., Valea F., French D.L.: “Correlation of the in situ detection of polymerase chain reactionamplified metalloproteinase complementary DNAs and their in-hibitors with prognosis in cervical carcinoma”. Cancer Res., 1995, 55, 267.

[38] Chebassier N., Leroy S., Tenaud I., Knol A.C., Dreno B.: “Overex-pression of MMP-2 and MMP-9 in squamous cell carcinomas of immunosuppressed patients”. Arch. Dermatol. Res., 2002, 294, 124.

[39] Liang B., Wang S., Zhu X.G., Yu Y.X., Cui Z.R., Yu Y.Z.: “Increased expression of mitogen-activated protein kinase and its upstream reg-ulating signal in human gastric cancer”. World J. Gastroenterol., 2005, 11, 623.

[40] Guo X., Ma N., Wang J., Song J., Bu X., Cheng Y., et al.: “Increased p38-MAPK is responsible for chemotherapy resistance in human gastric cancer cells”. BMC Cancer, 2008, 8, 375.

[41] Whyte J., Bergin O., Bianchi A., McNally S., Martin F.: “Key sig-nalling nodes in mammary gland development and cancer. Mitogen-activated protein kinase signalling in experimental models of breast cancer progression and in mammary gland development”. Breast Cancer Res., 2009, 11, 209.

[42] Schubbert S., Shannon K., Bollag G.: “Hyperactive Ras in develop-mental disorders and cancer”. Nat. Rev. Cancer, 2007, 7, 295.

[43] Zhang C., Zhu H., Yang X., Lou J., Zhu D., Lu W., et al.: “TP53 and p38 MAPK pathways are involved in MONCPT-induced cell cycle G2/M arrest in human nonsmall cell lung cancer A549”. J. Cancer Res. Clin. Oncol., 2010, 136, 437.

[44] Uzgare A.R., Kaplan P.J., Greenberg N.M.: “Differential expression and/or activation of P38MAPK, erk1/2, and jnk during the initiation and progression of prostate cancer”. Prostate, 2003, 55, 128.

[45] Reddy K.B., Nabha S.M., Atanaskova N.: “Role of MAP kinase in tumor progression and invasion”. Cancer Metastasis Rev., 2003, 22, 395.

[46] Wagner E.F., Nebreda A.R.: “Signal integration by JNK and p38 MAPK pathways in cancer development”. Nat. Rev. Cancer, 2009, 9, 537.

[47] Minden A., Lin A., Claret F.X., Abo A., Karin M.: “Selective activation of the JNK signaling cascade and c-Jun transcriptional activity by the small GTPases Rac and Cdc42Hs”. Cell, 1995, 81, 1147.

[48] Oktay M., Wary K.K., Dans M., Birge R.B., Giancotti F.G.: “Integrin-mediated activation of focal adhesion kinase is required for signaling to Jun NH2-terminal kinase and progression through the G1 phase of the cell cycle”. J. Cell. Biol., 1999, 145, 1461.

[49] Simon C., Hicks M.J., Nemechek A.J., Mehta R., O’Malley B.W. Jr., Goepfert H., et al.: “PD 098059, an inhibitor of ERK1 activation, attenuates the in vivo invasiveness of head and neck squamous cell carcinoma”. Br. J. Cancer, 1999, 80, 1412.

[50] Hubalek M.M., Widschwendter A., Erdel M., Gschwendtner A., Fiegl H.M., Muller H.M., et al.: “Cyclin E dysregulation and chromosomal instability in endometrial cancer”. Oncogene, 2004, 23, 4187.

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