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BMC Cancer
Published in: BMC Cancer 1/2016

Open Access 01-12-2016 | Research article

Relevance of miR-21 in regulation of tumor suppressor gene PTEN in human cervical cancer cells

Authors: Oscar Peralta-Zaragoza, Jessica Deas, Angélica Meneses-Acosta, Faustino De la O-Gómez, Gloria Fernández-Tilapa, Claudia Gómez-Cerón, Odelia Benítez-Boijseauneau, Ana Burguete-García, Kirvis Torres-Poveda, Victor Hugo Bermúdez-Morales, Vicente Madrid-Marina, Mauricio Rodríguez-Dorantes, Alfredo Hidalgo-Miranda, Carlos Pérez-Plasencia

Published in: BMC Cancer | Issue 1/2016

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Abstract

Background

Expression of the microRNA miR-21 has been found to be altered in almost all types of cancers and it has been classified as an oncogenic microRNA or oncomir. Due to the critical functions of its target proteins in various signaling pathways, miR-21 is an attractive target for genetic and pharmacological modulation in various cancers. Cervical cancer is the second most common cause of death from cancer in women worldwide and persistent HPV infection is the main etiologic agent. This malignancy merits special attention for the development of new treatment strategies. In the present study we analyze the role of miR-21 in cervical cancer cells.

Methods

To identify the downstream cellular target genes of upstream miR-21, we silenced endogenous miR-21 expression in a cervical intraepithelial neoplasia-derived cell lines using siRNAs. The effect of miR-21 on gene expression was assessed in cervical cancer cells transfected with the siRNA expression plasmid pSIMIR21. We identified the tumor suppressor gene PTEN as a target of miR-21 and determined the mechanism of its regulation throughout reporter construct plasmids. Using this model, we analyzed the expression of miR-21 and PTEN as well as functional effects such as autophagy and apoptosis induction.

Results

In SiHa cells, there was an inverse correlation between miR-21 expression and PTEN mRNA level as well as PTEN protein expression in cervical cancer cells. Transfection with the pSIMIR21 plasmid increased luciferase reporter activity in construct plasmids containing the PTEN-3′-UTR microRNA response elements MRE21-1 and MRE21-2. The role of miR-21 in cell proliferation was also analyzed in SiHa and HeLa cells transfected with the pSIMIR21 plasmid, and tumor cells exhibited markedly reduced cell proliferation along with autophagy and apoptosis induction.

Conclusions

We conclude that miR-21 post-transcriptionally down-regulates the expression of PTEN to promote cell proliferation and cervical cancer cell survival. Therefore, it may be a potential therapeutic target in gene therapy for cervical cancer.
Literature
1.
Valencia-Sanchez MA, Liu J, Hannon GJ, Parker R. Control of translation and mRNA degradation by miRNAs and siRNAs. Genes Dev. 2006;20:515–24.CrossRefPubMed
2.
Chan JA, Krichevsky AM, Kosik KS. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells. Cancer Res. 2005;65:6029–33.CrossRefPubMed
3.
Fabbri M, Campiglio M, Ménard S, Palazzo JP, Rosenberg A, Musiani P, et al. MicroRNA gene expression deregulation in human breast cancer. Cancer Res. 2005;65(16):7065–70.CrossRefPubMed
4.
Seike M, Goto A, Okano T, Bowman ED, Schetter AJ, Horikawa I, et al. MiR-21 is an EGFR-regulated anti-apoptotic factor in lung cancer in never-smokers. Proc Natl Acad Sci U S A. 2009;106(29):12085–90.CrossRefPubMedPubMedCentral
5.
Hiyoshi Y, Kamohara H, Karashima R, Sato N, Imamura Y, Nagai Y, et al. MicroRNA-21 regulates the proliferation and invasion in esophageal squamous cell carcinoma. Clin Cancer Res. 2009;15(6):1915–22.CrossRefPubMed
6.
Chan SH, Wu CW, Li AF, Chi CW, Lin WC. MiR-21 microRNA expression in human gastric carcinomas and its clinical association. Anticancer Res. 2008;28:907–11.PubMed
7.
Meng F, Henson R, Wehbe-Janek H, Ghoshal K, Jacob ST, Patel T. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer. Gastroenterology. 2007;133:647–58.CrossRefPubMedPubMedCentral
8.
Meng F, Henson R, Lang M, Wehbe H, Maheshwari S, Mendell JT, et al. Involvement of human Micro-RNA in growth and response to chemotherapy in human cholangiocarcinoma cell lines. Gastroenterology. 2006;130(7):2113–29.CrossRefPubMed
9.
Moriyama T, Ohuchida K, Mizumoto K, Yu J, Sato N, Nabae T, et al. MicroRNA-21 modulates biological functions of pancreatic cancer cells including their proliferation, invasion, and chemoresistance. Mol Cancer Ther. 2009;8(5):1067–74.CrossRefPubMed
10.
Volinia S, Calin GA, Liu CG, Ambs S, Cimmino A, Petrocca F, et al. A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci U S A. 2006;103(7):2257–61.CrossRefPubMedPubMedCentral
11.
Catto JW, Miah S, Owen HC, Bryant H, Myers K, Dudziec E, et al. Distinct microRNA alterations characterize highand low-grade bladder cancer. Cancer Res. 2009;69(21):8472–81.CrossRefPubMedPubMedCentral
12.
Iorio MV, Visone R, Di Leva G, Donati V, Petrocca F, Casalini P, et al. MicroRNA signatures in human ovarian cancer. Cancer Res. 2007;67(18):8699–707.CrossRefPubMed
13.
Yamanaka Y, Tagawa H, Takahashi N, Watanabe A, Guo YM, Iwamoto K, et al. Aberrant overexpression of microRNAs activate AKT signaling via downregulation of tumor suppressors in NK-cell lymphoma/leukemia. Blood. 2009;114(15):3265–75.CrossRefPubMed
14.
Liu M, Wu H, Liu T, Li Y, Wang F, Wan H, et al. Regulation of the cell cycle gene, BTG2, by miR-21 in human laryngeal carcinoma. Cell Res. 2009;19(7):828–37.CrossRefPubMed
15.
Li J, Huang H, Sun L, Yang M, Pan C, Chen W, et al. MiR-21 indicates poor prognosis in tongue squamous cell carcinomas as an apoptosis inhibitor. Clin Cancer Res. 2009;15(12):3998–4008.CrossRefPubMed
16.
Medina PP, Nolde M, Slack FJ. OncomiR addiction in an in vivo model of microRNA-21-induced pre-B-cell lymphoma. Nature. 2010;467:86–90.CrossRefPubMed
17.
Ramachandra RK, Salem M, Gahr S, Rexroad 3rd CE, Yao J. Cloning and characterization of microRNAs from rainbow trout (Oncorhynchus mykiss): their expression during early embryonic development. BMC Dev Biol. 2008;8:41.CrossRefPubMedPubMedCentral
18.
19.
Gabriely G, Wurdinger T, Kesari S, Esau CC, Burchard J, Linsley PS, et al. MicroRNA 21 promotes glioma invasion by targeting matrix metalloproteinase regulators. Mol Cell Biol. 2008;28:5369–80.CrossRefPubMedPubMedCentral
20.
Qian B, Katsaros D, Lu L, Preti M, Durando A, Arisio R, et al. High miR-21 expression in breast cancer associated with poor disease-free survival in early stage disease and high TGFbeta1. Breast Cancer Res Treat. 2009;117(1):131–40.CrossRefPubMed
21.
22.
Markou A, Tsaroucha EG, Kaklamanis L, Fotinou M, Georgoulias V, Lianidou ES. Prognostic value of mature microRNA-21 and microRNA-205 overexpression in non-small cell lung cancer by quantitative realtime RT-PCR. Clin Chem. 2008;54(10):1696–704.CrossRefPubMed
23.
Xie Y, Todd NW, Liu Z, Zhan M, Fang H, Peng H, et al. Altered miRNA expression in sputum for diagnosis of non-small cell lung cancer. Lung Cancer. 2010;67(2):170–6.CrossRefPubMedPubMedCentral
24.
Gao W, Yu Y, Cao H, Shen H, Li X, Pan S, et al. Deregulated expression of miR-21, miR-143 and miR-181a in non- small cell lung cancer is related to clinicopathologic characteristics or patient prognosis. Biomed Pharmacother. 2010;64(6):399–408.CrossRefPubMed
25.
Hirata Y, Murai N, Yanaihara N, Saito M, Saito M, Urashima M, et al. MicroRNA-21 is a candidate driver gene for 17q23-25 amplification in ovarian clear cell carcinoma. BMC Cancer. 2014;14:799.CrossRefPubMedPubMedCentral
26.
Selcuklu SD, Donoghue MT, Spillane C. MiR-21 as a key regulator of oncogenic processes. Biochem Soc Trans. 2009;37:918–25.CrossRefPubMed
27.
Papagiannakopoulos T, Shapiro A, Kosik KS. MicroRNA-21 targets a network of key tumor-suppressive pathways in glioblastoma cells. Cancer Res. 2008;68:8164–72.CrossRefPubMed
28.
Zhang JG, Wang JJ, Zhao F, Liu Q, Jiang K, Yang GH. MicroRNA-21 (miR-21) represses tumor suppressor PTEN and promotes growth and invasion in non-small cell lung cancer (NSCLC). Clin Chim Acta. 2010;411(11–12):846–52.CrossRefPubMed
29.
Asangani IA, Rasheed SA, Nikolova DA, Leupold JH, Colburn NH, Post S, et al. MicroRNA-21 (miR-21) posttranscriptionally down-regulates tumor suppressor PDCD4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene. 2007;27:2128–36.CrossRefPubMed
30.
Zhu S, Wu H, Wu F, Nie D, Sheng S, Mo YY. MicroRNA-21 targets tumor suppressor genes in invasion and metastasis. Cell Res. 2008;18:350–9.CrossRefPubMed
31.
Zhu S, Si M-L, Wu H, Mo YY. MicroRNA-21 targets the tumor suppressor gene tropomyosin 1 (TPM1). J Biol Chem. 2007;282:14328–36.CrossRefPubMed
32.
Lui WO, Pourmand N, Patterson BK, Fire A. Patterns of known and novel small RNAs in human cervical cancer. Cancer Res. 2007;67(13):6031–43.CrossRefPubMed
33.
Yao T, Lin Z. MiR-21 is involved in cervical squamous cell tumorigenesis and regulates CCL20. Biochim Biophys Acta. 2012;1822(2):248–60.CrossRefPubMed
34.
Gocze K, Gombos K, Kovacs K, Juhasz K, Gocze P, Kiss I. MicroRNA Expressions in HPV-induced Cervical Dysplasia and Cancer. Anticancer Res. 2015;35(1):523–30.PubMed
35.
Bruni L, Barrionuevo-Rosas L, Albero G, Aldea M, Serrano B, Valencia S, Brotons M, Mena M, Cosano R, Muñoz J, Bosch FX, de Sanjosé S, Castellsagué X. ICO Information Centre on HPV and Cancer (HPV Information Centre). Human Papillomavirus and Related Diseases in the World. Summary Report. Dec 18 2014; p 227.
36.
Zur Hausen H. Papillomaviruses and cancer: from basic studies to clinical application. Nat Rev Cancer. 2002;2:342–50.CrossRefPubMed
37.
Steller MA. Human papillomavirus, it’s genes…and cancer vaccines. Cancer Cell. 2003;3(1):7–8.CrossRefPubMed
38.
Saito-Ramalho A, Dantas-Lopes A, Talans A, Parlato-Sakiyama BY, Stelko-Pereira GL, Hoff PM, et al. Molecular targets for therapeutic interventions in human papillomavirus-related cancers. Oncol Rep. 2010;24(6):1419–26.PubMed
39.
Pim D, Banks L. Interaction of viral oncoproteins with cellular target molecules: infection with high-risk vs low-risk human papillomaviruses. APMIS. 2010;118(6–7):471–93.CrossRefPubMed
40.
41.
42.
43.
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.CrossRefPubMed
44.
Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, et al. Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 2005;33(20), e179.CrossRefPubMedPubMedCentral
45.
46.
Thorland EC, Myers SL, Gostout BS, Smith DI. Common fragile sites are preferential targets for HPV16 integrations in cervical tumors. Oncogene. 2003;22:1225–37.CrossRefPubMed
47.
Sansal I, Sellers WR. The biology and clinical relevance of the PTEN tumor suppressor pathway. J Clin Oncol. 2004;22(14):2954–63.CrossRefPubMed
48.
Zhang BG, Li JF, Yu BQ, Zhu ZG, Liu BY, Yan M. MicroRNA-21 promotes tumor proliferation and invasion in gastric cancer by targeting PTEN. Oncol Rep. 2012;4:1019–26.
49.
Chen B, Chen X, Wu X, Wang X, Wang Y, Lin TY, Kurata J, et al. Disruption of microRNA-21 by TALEN leads to diminished cell transformation and increased expression of cell-environment interaction genes. Cancer Lett. 2015;356(2 Pt B):506–16.CrossRefPubMedPubMedCentral
50.
Tay Y, Song SJ, Pandolfi PP. The Lilliputians and the Giant: An Emerging Oncogenic microRNA Network that Suppresses the PTEN Tumor Suppressor In Vivo. Microrna. 2013;2(2):127–36.CrossRefPubMed
51.
Stewart AL, Mhashilkar AM, Yang XH, Ekmekcioglu S, Saito Y, Sieger K, Schrock R, et al. PI3K blockade by Ad-PTEN inhibits invasion and induces apoptosis in radial growth phase and metastatic melanoma cells. Mol Med. 2002;8:451–61.PubMedPubMedCentral
52.
Tamura M, Gu J, Matsumoto K, Aota S, Parsons R, Yamada KM. Inhibition of cell migration, spreading, and focal adhesions by tumor suppressor PTEN. Science. 1998;280:614–7.CrossRef
53.
Castellino RC, Durden DL. Mechanisms of disease: the PI3K-Akt-PTEN signaling node - an intercept point for the control of angiogenesis in brain tumors. Nat Clin Pract Neurol. 2007;3:682–93.CrossRefPubMed
54.
Zhou X, Ren Y, Moore L, Mei M, You Y, Xu P, et al. Downregulation of miR-21 inhibits EGFR pathway and suppresses the growth of human glioblastoma cells independent of PTEN status. Lab Investig. 2010;90:144–55.CrossRefPubMed
55.
Shishodia G, Verma G, Srivastava Y, Mehrotra R, Das BC, Bharti AC. Deregulation of microRNAs Let-7a and miR-21 mediate aberrant STAT3 signaling during human papillomavirus-induced cervical carcinogenesis: role of E6 oncoprotein. BMC Cancer. 2014;14:996.CrossRefPubMedPubMedCentral
56.
Shishodia G, Shukla S, Srivastava Y, Masaldan S, Mehta S, Bhambhani S, Sharma S, Mehrotra R, Das BC, Bharti AC. Alterations in microRNAs miR-21 and let-7a correlate with aberrant STAT3 signaling and downstream effects during cervical carcinogenesis. Mol Cancer. 2015;14:116.CrossRefPubMedPubMedCentral
57.
Wang N, Xu Z, Wang K, Zhu M, Li Y. Construction and analysis of regulatory genetic networks in cervical cancer based on involved microRNAs, target genes, transcription factors and host genes. Oncol Lett. 2014;7(4):1279–83.PubMedPubMedCentral
58.
Suzuki HI, Yamagata K, Sugimoto K, Iwamoto T, Kato S, Miyazono K. Modulation of microRNA processing by p53. Nature. 2009;460:529–33.CrossRefPubMed
59.
60.
61.
Maiuri CM, Galluzzi L, Morselli E, Kepp O, Malik SA, Kroemer G. Autophagy regulation by p53. Curr Opin Cell Biol. 2009;22:1–5.
62.
Frankel LB, Christoffersen NR, Jacobsen A, Lindow M, Krogh A, Lund AH. Programmed cell death 4 (PDCD4) is an important functional target of the microRNA miR-21 in breast cancer cells. J Biol Chem. 2008;283:1026–33.CrossRefPubMed
63.
Bornachea O, Santos M, Martínez-Cruz AB, García-Escudero R, Dueñas M, Costa C, et al. EMT and induction of miR-21 mediate metastasis development in Trp53-deficient tumours. Sci Rep. 2012;2:434.CrossRefPubMedPubMedCentral
64.
Xu J, Zhang W, Lv Q, Zhu D. Overexpression of miR-21 promotes the proliferation and migration of cervical cancer cells via the inhibition of PTEN. Oncol Rep. 2015;33(6):3108–16.PubMed
65.
Li DM, Sun H. TEP1, encoded by a candidate tumor suppressor locus, is a novel protein tyrosine phosphatase regulated by transforming growth factor beta. Cancer Res. 1997;57:2124–9.PubMed
66.
Lee SB, Tong SY, Kim JJ, Um SJ, Park JS. Caspase-independent autophagic cytotoxicity in etoposide-treated CaSki cervical carcinoma cells. DNA Cell Biol. 2007;26(10):713–20.CrossRefPubMed
67.
Yang C, Kaushal V, Shah SV, Kaushal GP. Autophagy is associated with apoptosis in cisplatin injury to renal tubular epithelial cells. Am J Physiol Renal Physiol. 2008;294(4):F777–87.CrossRefPubMed
68.
69.
70.
Tricoli JV, Jacobson JW. MicroRNA: Potential for cancer detection, diagnosis, and prognosis. Cancer Res. 2007;67:4553–5.CrossRefPubMed
Metadata
Title
Relevance of miR-21 in regulation of tumor suppressor gene PTEN in human cervical cancer cells
Authors
Oscar Peralta-Zaragoza
Jessica Deas
Angélica Meneses-Acosta
Faustino De la O-Gómez
Gloria Fernández-Tilapa
Claudia Gómez-Cerón
Odelia Benítez-Boijseauneau
Ana Burguete-García
Kirvis Torres-Poveda
Victor Hugo Bermúdez-Morales
Vicente Madrid-Marina
Mauricio Rodríguez-Dorantes
Alfredo Hidalgo-Miranda
Carlos Pérez-Plasencia
Publication date
01-12-2016
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2016
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/s12885-016-2231-3

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