MicroRNA-200c and microRNA-31 regulate proliferation, colony formation, migration and invasion in serous ovarian cancer

Journal of Ovarian Research

Norfilza Mohd Mokhtar²  Mohammad Manir Hossain Mollah¹  Reena Rahayu MdZin³  Sazuita Saidin¹  Nurul Syakima Ab Mutalib¹  Saiful Effendi Syafruddin¹  Rahman Jamal¹  Fateen Farhana Ibrahim¹ 

[1] UKM Medical Molecular Biology Institute, Universiti Kebangsaan Malaysia, Jalan Yaa′cob Latiff, Bandar Tun Razak, Kuala Lumpur, Malaysia;Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia Medical Center, Jalan Yaacob Latiff, Bandar Tun Razak, Kuala Lumpur, Malaysia;Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff, Kuala Lumpur, Malaysia

关键词: Invasion;    Migration;    In situ hybridization;    Serous ovarian cancer;    miR-31;    miR-200c;    MicroRNA;   

Others  :  1224164
DOI  :  10.1186/s13048-015-0186-7

 received in 2015-02-22, accepted in 2015-08-06,  发布年份 2015

【 摘 要 】

Background

Serous epithelial ovarian cancer (SEOC) is a highly metastatic disease and its progression has been implicated with microRNAs. This study aimed to identify the differentially expressed microRNAs in Malaysian patients with SEOC and examine the microRNAs functional roles in SEOC cells.

Methods

Twenty-two SEOC and twenty-two normal samples were subjected to miRNA expression profiling using the locked nucleic acid (LNA) quantitative real-time PCR (qPCR). The localization of miR-200c was determined via LNA in situ hybridization (ISH). Functional analysis of miR-200c and miR-31 on cell proliferation, migration and invasion and clonogenic cell survival were assessed in vitro. The putative target genes of the two miRNAs were predicted by miRWalk program and expression of the target genes in SEOC cell lines was validated.

Results

The miRNA expression profiling revealed thirty-eight significantly dysregulated miRNAs in SEOC compared to normal ovarian tissues. Of these, eighteen were up-regulated whilst twenty miRNAs were down-regulated. We observed chromogenic miR-200c-ISH signal predominantly in the cytoplasmic compartment of both epithelial and inflammatory cancer cells. Re-expression of miR-200c significantly increased the cell proliferation and colony formation but reduced the migration and invasion of SEOC cells. In addition, miR-200c expression was inversely proportionate with the expression of deleted in liver cancer-1 (DLC-1) gene. Over-expression of miR-31 in SEOC cells resulted in decreased cell proliferation, clonogenic potential, cell migration and invasion. Meanwhile, miR-31 gain-of-function led to the down-regulation of AF4/FMR2 family member 1 (AFF1) gene.

Conclusions

These data suggested that miR-200c and miR-31 may play roles in the SEOC metastasis biology and could be considered as promising targets for therapeutic purposes.

【 授权许可】

   
2015 Ibrahim et al.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Fig. 6.

【 参考文献 】

• [1]Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D. Global cancer statistics. CA Cancer J Clin. 2011; 61:69-90.
• [2]Zainal A, Nor Saleha I. National Cancer Registry Report: Malaysia Cancer Statistics – Data and Figure 2007. National Cancer Registry Ministry of Health Malaysia, Malaysia; 2011.
• [3]Prat J. Ovarian carcinomas: five distinct diseases with different origins, genetic alterations, and clinicopathological features. Virchows Arch. 2012; 460:237-49.
• [4]Ng JS, Low JJ, Ilancheran A. Epithelial ovarian cancer. Best Pract Res Clin Obstetrics Gynaecol. 2012; 26:337-45.
• [5]Malpica A, Deavers MT, Lu K, Bodurka DC, Atkinson EN, Gershenson DM, Silva EG. Grading ovarian serous carcinoma using a two-tier system. Am J Surg Pathol. 2004; 28:496-504.
• [6]Lengyel E. Ovarian cancer development and metastasis. Am J Pathol. 2010; 177:1053-64.
• [7]Chen CZ, Li L, Lodish HF, Bartel DP. MicroRNAs modulate hematopoietic lineage differentiation. Science. 2004; 303:83-6.
• [8]Garzon R, Marcucci G, Croce CM. Targeting microRNAs in cancer: rationale, strategies and challenges. Nat Rev Drug Discov. 2010; 9:775-89.
• [9]Kozomara A, Griffiths-Jones S. miRBase: annotating high confidence microRNAs using deep sequencing data (1). Nucleic Acids Res. 2013; 42:D68-73.
• [10]Jansson MD, Lund AH. MicroRNA and cancer. Mol Oncol. 2012; 6:590-610.
• [11]Cochrane DR, Howe EN, Spoelstra NS, Richer JK. Loss of miR-200c: A Marker of Aggressiveness and Chemoresistance in Female Reproductive Cancers. J Oncol. 2010; 2010:1-2.
• [12]Hurteau GJ, Carlson JA, Spivack SD, Brock GJ. Overexpression of the microRNA hsa-miR-200c leads to reduced expression of transcription factor 8 and increased expression of E-cadherin. Cancer Res. 2007; 67:7972-6.
• [13]Pacurari M, Addison JB, Bondalapati N, Wan YW, Luo D, Qian Y, Castranova V, Ivanov AV, Guo NL. The microRNA-200 family targets multiple non-small cell lung cancer prognostic markers in H1299 cells and BEAS-2B cells. Int J Oncol. 2013; 43:548-60.
• [14]Yang MH, Yu J, Chen N, Wang XY, Liu XY, Wang S, Ding YQ, Sarkar FH. Elevated microRNA-31 expression regulates colorectal cancer progression by repressing its target gene SATB2. PLoS One. 2013; 8: Article ID e85353
• [15]Mitamura T, Watari H, Wang L, Kanno H, Kitagawa M, Hassan MK, Kimura T, Tanino M, Nishihara H, Tanaka S et al.. microRNA 31 functions as an endometrial cancer oncogene by suppressing Hippo tumor suppressor pathway. Mol Cancer. 2014; 13:97. BioMed Central Full Text
• [16]Valastyan S, Reinhardt F, Benaich N, Calogrias D, Szász AM, Wang ZC, Brock JE, Richardson AL, Weinberg RA. A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell. 2009; 137:1032-46.
• [17]Hua D, Ding D, Han X, Zhang W, Zhao N, Foltz G, Lan Q, Huang Q, Lin B. Human miR-31 targets radixin and inhibits migration and invasion of glioma cells. Oncol Rep. 2012; 27:700-6.
• [18]Schaefer A, Jung M, Mollenkopf HJ, Wagner I, Stephan C, Jentzmik F, Miller K, Lein M, Kristiansen G, Jung K et al.. Diagnostic and prognostic implications of microRNA profiling in prostate carcinoma. Int J Cancer. 2010; 126:1166-76.
• [19]Schmittgen TD. miR-31: a master regulator of metastasis? Future Oncol. 2010; 6:17-20.
• [20]Yuan BZ, Miller MJ, Keck CL, Zimonjic DB, Thorgeirsson SS, Popescu NC. Cloning, characterization, and chromosomal localization of a gene frequently deleted in human liver cancer (DLC-1) homologous to rat RhoGAP. Cancer Res. 1998; 58:2196-9.
• [21]Healy KD, Hodgson L, Kim TY, Shutes A, Maddileti S, Juliano RL, Hahn KM, Harden TK, Bang YJ, Der CJ et al.. DLC-1 suppresses non-small cell lung cancer growth and invasion by RhoGAP-dependent and independent mechanisms. Mol Carcinog. 2008; 47:326-37.
• [22]Plaumann M, Seitz S, Frege R, Estevez-Schwarz L, Scherneck S. Analysis of DLC-1 expression in human breast cancer. J Cancer Res Clin Oncol. 2003; 129:349-54.
• [23]Wu PP, Jin YL, Shang YF, Jin Z, Wu P, Huang PL. Restoration of DLC1 gene inhibits proliferation and migration of human colon cancer HT29 cells. Ann Clin Lab Sci. 2009; 39:263-9.
• [24]Ren F, Shi H, Zhang G, Zhang R. Expression of deleted in liver cancer 1 and plasminogen activator inhibitor 1 protein in ovarian carcinoma and their clinical significance. J Exp Clin Cancer Res. 2013; 32:60. BioMed Central Full Text
• [25]Elgaaen BV, Olstad OK, Haug KB, Brusletto B, Sandvik L, Staff AC, Gautvik KM, Davidson B. Global miRNA expression analysis of serous and clear cell ovarian carcinomas identifies differentially expressed miRNAs including miR-200c-3p as a prognostic marker. BMC Cancer. 2014; 14:80. BioMed Central Full Text
• [26]Iorio MV, Visone R, Di Leva G, Donati V, Petrocca F, Casalini P, Taccioli C, Volinia S, Liu CG, Alder H et al.. MicroRNA signatures in human ovarian cancer. (1). Cancer Res. 2007; 67:8699-707.
• [27]Lee CH, Subramanian S, Beck AH, Espinosa I, Senz J, Zhu SX, Huntsman D, van de Rijn M, Gilks CB, Toland AE et al.. MicroRNA profiling of BRCA1/2 mutation-carrying and non-mutation-carrying high-grade serous carcinomas of ovary. PLoS One. 2009; 4: Article ID e7314
• [28]Nam EJ, Yoon H, Kim SW, Kim H, Kim YT, Kim JH, Kim JW, Kim S. MicroRNA expression profiles in serous ovarian carcinoma. Clin Cancer Res. 2008; 14:2690-5.
• [29]Creighton CJ, Fountain MD, Yu Z, Nagaraja AK, Zhu H, Khan M, Olokpa E, Zariff A, Gunaratne PH, Matzuk MM et al.. Molecular profiling uncovers a p53-associated role for microRNA-31 in inhibiting the proliferation of serous ovarian carcinomas and other cancers. Cancer Res. 2010; 70:1906-15.
• [30]Wyman SK, Parkin RK, Mitchell PS, Fritz BR, O'Briant K, Godwin AK, Urban N, Drescher CW, Knudsen BS, Tewari M et al.. Repertoire of microRNAs in epithelial ovarian cancer as determined by next generation sequencing of small RNA cDNA libraries. PLoS One. 2009; 4: Article ID e5311
• [31]Nuovo GJ. In situ detection of microRNAs in paraffin embedded, formalin fixed tissues and the co-localization of their putative targets. Methods. 2010; 52:307-15.
• [32]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.
• [33]Kan CWS, Hahn MA, Gard GB, Maidens J, Huh JY, Marsh DJ, Howell VM. Elevated levels of circulating microRNA-200 family members correlate with serous epithelial ovarian cancer. BMC Cancer. 2012; 12:627. BioMed Central Full Text
• [34]Liu H, Shi H, Hao Y, Zhao G, Yang X, Wang Y, Li M, Liu M. Effect of FAK, DLC-1 gene expression on OVCAR-3 proliferation. Mol Biol Rep. 2012; 39:10665-70.
• [35]Goodison S, Yuan J, Sloan D, Kim R, Li C, Popescu NC, Urquidi V. The RhoGAP protein DLC-1 functions as a metastasis suppressor in breast cancer cells. Cancer Res. 2005; 65:6042-53.
• [36]Yu J, Ohuchida K, Mizumoto K, Sato N, Kayashima T, Fujita H, Nakata K, Tanaka M. MicroRNA, hsa-miR-200c, is an independent prognostic factor in pancreatic cancer and its upregulation inhibits pancreatic cancer invasion but increases cell proliferation. Mol Cancer. 2010; 9:169. BioMed Central Full Text
• [37]Hur K, Toiyama Y, Takahashi M, Balaguer F, Nagasaka T, Koike J, Hemmi H, Koi M, Boland CR, Goel A et al.. MicroRNA-200c modulates epithelial-to-mesenchymal transition (EMT) in human colorectal cancer metastasis. Gut. 2013; 62:1315-26.
• [38]Luo Z, Lin C, Shilatifard A. The super elongation complex (SEC) family in transcriptional control. Nat Rev Mol Cell Biol. 2012; 13:543-7.
• [39]Dou L, Zheng D, Li J, Li Y, Gao L, Wang L, Yu L. Methylation-mediated repression of microRNA-143 enhances MLL-AF4 oncogene expression. Oncogene. 2012; 31:507-17.