ABSTRACT
Purpose
MicroRNA-101 (miR-101) expression is negatively associated with tumor growth and proliferation in several solid epithelial cancers. Enhancer of zeste homolog 2 (EzH2) appears to be a functional target of miR-101. We explore the role of miR-101 and its interaction with EzH2 in epithelial ovarian carcinoma (EOC).
Methods
In situ hybridization (ISH) for miR-101 was performed on EOC patient tissues and normal controls. EOC cell lines were transfected with miR-101 and subjected to growth analysis and clonogenic assays. Cell motility was assessed by Boyden chamber and wound-healing assays. P21waf1/cip1 and EzH2 interaction was assessed by Chromatin Immunoprecipitation (ChIP) assay in MDAH-2774 cells. SCID mice were assessed for tumor burden after injection with miR-101 or control vector-treated MDAH-2774 cells.
Results
ISH analysis revealed a decrease in miR-101 expression in EOC compared with normal tissue. MiR-101 re-expression in EOC cell lines resulted in increased apoptosis, decreased cellular proliferation, invasiveness, and reduced growth of tumor xenografts. CHIP assays revealed that re-expression of miR-101 inhibited the interaction of EzH2 with p21waf1/cip1 promoter.
Conclusions
MiR-101 re-expression appears to have antitumor effects, providing a better understanding of the role of miR-101 in EOC.
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REFERENCES
Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun M. Cancer statistics, 2009. CA: A Cancer J Clinicians. 2009;59:225.
McGuire W, Hoskins W, Brady M, Kucera P, Partridge E, Look K, et al. Cyclophosphamide and cisplatin compared with paclitaxel and cisplatin in patients with stage III and stage IV ovarian cancer. New Engl J Med. 1996;334:1.
Aghajanian C. Clinical update: novel targets in gynecologic malignancies, Vol. 31. Elsevier; 2004. p. 22–26.
Trimbosand J, Timmers P. Chemotherapy for early ovarian cancer. Curr Opin Obstet Gynecol. 2004;16:43.
Valencia-Sanchez M, Liu J, Hannon G, Parker R. Control of translation and mRNA degradation by miRNAs and siRNAs. Gene Dev. 2006;20:515.
Esquela-Kerscherand A, Slack F. OncomirsómicroRNAs with a role in cancer. Nat Rev Canc. 2006;6:259.
Calin G, Sevignani C, Dumitru C, Hyslop T, Noch E, Yendamuri S, et al. Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci. 2004;101:2999.
Zhang L, Huang J, Yang N, Greshock J, Megraw M, Giannakakis A, et al. microRNAs exhibit high frequency genomic alterations in human cancer. Proc Natl Acad Sci. 2006;103:9136.
Lu L, Katsaros D, Rigault de la Longrais I, Sochirca O, Yu H. Hypermethylation of let-7a-3 in epithelial ovarian cancer is associated with low insulin-like growth factor-II expression and favorable prognosis. Canc Res. 2007;67:10117.
Dahiya N, Sherman-Baust C, Wang T, Davidson B, Shih I, Zhang Y, et al. MicroRNA expression and identification of putative miRNA targets in ovarian cancer. PLos One. 2008;3.
Friedman J, Liang G, Liu C, Wolff E, Tsai Y, Ye W, et al. The Putative Tumor Suppressor microRNA-101 Modulates the Cancer Epigenome by Repressing the Polycomb Group Protein EZH2. Canc Res. 2009;69:2623.
Varambally S, Cao Q, Mani RS, Shankar S, Wang X, Ateeq B, et al. Genomic loss of microRNA-101 leads to overexpression of histone methyltransferase EZH2 in cancer. Science. 2008;322:1695–9.
Malpica A, Deavers M, Lu K, Bodurka D, Atkinson E, Gershenson D, et al. Grading ovarian serous carcinoma using a two-tier system. Am J Surg Pathol. 2004;28:496.
Bachmann IM, Halvorsen OJ, Collett K, Stefansson IM, Straume O, Haukaas SA, et al. EZH2 expression is associated with high proliferation rate and aggressive tumor subgroups in cutaneous melanoma and cancers of the endometrium, prostate, and breast. J Clin Oncol. 2006;24:268–73.
Kleer CG, Cao Q, Varambally S, Shen R, Ota I, Tomlins SA, et al. EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells. Proc Natl Acad Sci USA. 2003;100:11606–11.
Raman JD, Mongan NP, Tickoo SK, Boorjian SA, Scherr DS, Gudas LJ, et al. Increased expression of the polycomb group gene, EZH2, in transitional cell carcinoma of the bladder. Clin Canc Res. 2005;11:8570–6.
Varambally S, Dhanasekaran S, Zhou M, Barrette T, Kumar-Sinha C, Sanda M, et al. The polycomb group protein EZH2 is involved in progression of prostate cancer. Nature. 2002;419:624–9.
Krek A, Grün D, Poy MN, Wolf R, Rosenberg L, Epstein EJ, et al. Combinatorial microRNA target predictions. Nat Genet. 2005;37:495–500.
Reik W. Stability and flexibility of epigenetic gene regulation in mammalian development. Nature. 2007;447:425–32.
Wan M, Zweizig S, D’ablaing G, Zheng J, Velicescu M, Dubeau L. Three distinct regions of chromosome 6 are targets of loss of heterozygosity in human ovarian carcinomas. Int J Oncol. 1994;5:1043–8.
el-Deiry WS, Harper JW, O’Connor PM, Velculescu VE, Canman CE, Jackman J, et al. WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Canc Res. 1994;54:1169–74.
Bandres E, Agirre X, Ramirez N, Zarate R, Garcia-Foncillas J. MicroRNAs as cancer players: potential clinical and biological effects. DNA Cell Biol. 2007;26:273–82.
Cumminsand J, Velculescu V. Implications of micro-RNA profiling for cancer diagnosis. Oncogene. 2006;25:6220–7.
Zhang B, Pan X, Cobb GP, Anderson TA, Zhang B, Pan X, et al. microRNAs as oncogenes and tumor suppressors. Dev Biol. 2007;302:1–12.
Iorio M, Visone R, Di Leva G, Donati V, Petrocca F, Casalini P, et al. MicroRNA signatures in human ovarian cancer. Canc Res. 2007;67:8699.
Taylorand D, Gercel-Taylor C. MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol Oncol. 2008;110:13–21.
Yang H, Kong W, He L, Zhao J, O’Donnell J, Wang J, et al. MicroRNA expression profiling in human ovarian cancer: miR-214 induces cell survival and cisplatin resistance by targeting PTEN. Canc Res. 2008;68:425.
Zhang L, Volinia S, Bonome T, Calin G, Greshock J, Yang N, et al. Genomic and epigenetic alterations deregulate microRNA expression in human epithelial ovarian cancer. Proc Natl Acad Sci. 2008;105:7004.
Strillacci A, Griffoni C, Sansone P, Paterini P, Piazzi G, Lazzarini G, et al. MiR-101 downregulation is involved in cyclooxygenase-2 overexpression in human colon cancer cells. Exp Cell Res. 2009;315:1439–47.
Chao-Wei C, Yi H, Ka-Wai L, Lih-Chyang C, Hua-Chien C, Shu-Jen C, et al. PKC alpha mediated induction of miR-101 in human hepatoma HepG2 cells. J Biomed Sci. 2040;17.
Cao Q, Yu J, Dhanasekaran S, Kim J, Mani R, Tomlins S, et al. Repression of E-cadherin by the polycomb group protein EZH2 in cancer. Oncogene. 2008;27:7274–84.
Calinand G, Croce C. MicroRNA signatures in human cancers. Nat Rev Canc. 2006;6:857–66.
Ougolkov A, Bilim V, Billadeau D. Regulation of pancreatic tumor cell proliferation and chemoresistance by the histone methyltransferase enhancer of zeste homologue 2. Clin Canc Res. 2008;14:6790.
Haradaand K, Ogden G. An overview of the cell cycle arrest protein, p21WAF1. Oral Oncol. 2000;36:3–7.
Schmider A, Gee C, Friedmann W, Lukas J, Press M, Lichtenegger W, et al. p21 (WAF1/CIP1) protein expression is associated with prolonged survival but not with p53 expression in epithelial ovarian carcinoma. Gynecol Oncol. 2000;77:237–42.
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Assaad Semaan, Aamer M. Qazi, and Shelly Seward are equal contributors to the manuscript.
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Semaan, A., Qazi, A.M., Seward, S. et al. MicroRNA-101 Inhibits Growth of Epithelial Ovarian Cancer by Relieving Chromatin-Mediated Transcriptional Repression of p21waf1/cip1 . Pharm Res 28, 3079–3090 (2011). https://doi.org/10.1007/s11095-011-0547-x
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DOI: https://doi.org/10.1007/s11095-011-0547-x