Abstract
Rap1GAP is a critical tumor suppressor gene that is downregulated in multiple aggressive cancers, such as head and neck squamous cell carcinoma, melanoma and pancreatic cancer. However, the mechanistic basis of rap1GAP downregulation in cancers is poorly understood. By employing an integrative approach, we demonstrate polycomb-mediated repression of rap1GAP that involves Enhancer of Zeste Homolog 2 (EZH2), a histone methyltransferase in head and neck cancers. We further demonstrate that the loss of miR-101 expression correlates with EZH2 upregulation, and the concomitant downregulation of rap1GAP in head and neck cancers. EZH2 represses rap1GAP by facilitating the trimethylation of histone 3 at lysine 27, a mark of gene repression, and also hypermethylation of rap1GAP promoter. These results provide a conceptual framework involving a microRNA–oncogene–tumor suppressor axis to understand head and neck cancer progression.
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References
Altschuler DL, Ribeiro-Neto F . (1998). Mitogenic and oncogenic properties of the small G protein Rap1b. Proc Natl Acad Sci USA 95: 7475–7479.
Beke L, Nuytten M, Van Eynde A, Beullens M, Bollen M . (2007). The gene encoding the prostatic tumor suppressor PSP94 is a target for repression by the polycomb group protein EZH2. Oncogene 26: 4590–4595.
Bos JL, de Rooij J, Reedquist KA . (2001). Rap1 signalling: adhering to new models. Nat Rev Mol Cell Biol 2: 369–377.
Bracken AP, Helin K . (2009). Polycomb group proteins: navigators of lineage pathways led astray in cancer. Nat Rev Cancer 9: 773–784.
Cao Q, Yu J, Dhanasekaran SM, Kim JH, Mani RS, Tomlins SA et al. (2008). Repression of E-cadherin by the polycomb group protein EZH2 in cancer. Oncogene 27: 7274–7284.
Caron E, Self AJ, Hall A . (2000). The GTPase Rap1 controls functional activation of macrophage integrin alphaMbeta2 by LPS and other inflammatory mediators [In Process Citation]. Curr Biol 10: 974–978.
Chi P, Allis CD, Wang GG . (2010). Covalent histone modifications—miswritten, misinterpreted and mis-erased in human cancers. Nat Rev Cancer 10: 457–469.
Chin D, Boyle GM, Porceddu S, Theile DR, Parsons PG, Coman WB . (2006). Head and neck cancer: past, present and future. Expert Rev Anticancer Ther 6: 1111–1118.
Ernst T, Chase AJ, Score J, Hidalgo-Curtis CE, Bryant C, Jones AV et al. (2010). Inactivating mutations of the histone methyltransferase gene EZH2 in myeloid disorders. Nat Genet 42: 722–726.
Francis NJ, Kingston RE, Woodcock CL . (2004). Chromatin compaction by a polycomb group protein complex. Science 306: 1574–1577.
Friedman JM, Liang G, Liu CC, Wolff EM, Tsai YC, Ye W et al. (2009). The putative tumor suppressor microRNA-101 modulates the cancer epigenome by repressing the polycomb group protein EZH2. Cancer Res 69: 2623–2629.
Glazer CA, Chang SS, Ha PK, Califano JA . (2009). Applying the molecular biology and epigenetics of head and neck cancer in everyday clinical practice. Oral Oncol 45: 440–446.
He LR, Liu MZ, Li BK, Jia WH, Zhang Y, Liao YJ et al. (2010). High expression of EZH2 is associated with tumor aggressiveness and poor prognosis in patients with esophageal squamous cell carcinoma treated with definitive chemoradiotherapy. Int J Cancer 127: 138–147.
Herceg Z . (2007). Epigenetics and cancer: towards an evaluation of the impact of environmental and dietary factors. Mutagenesis 22: 91–103.
Hogan C, Serpente N, Cogram P, Hosking CR, Bialucha CU, Feller SM et al. (2004). Rap1 regulates the formation of E-cadherin-based cell-cell contacts. Mol Cell Biol 24: 6690–6700.
Kidani K, Osaki M, Tamura T, Yamaga K, Shomori K, Ryoke K et al. (2009). High expression of EZH2 is associated with tumor proliferation and prognosis in human oral squamous cell carcinomas. Oral Oncol 45: 39–46.
Kleer CG, Cao Q, Varambally S, Shen R, Ota I, Tomlins SA et al. (2003). EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells. Proc Natl Acad Sci USA 100: 11606–11611.
Kondo Y, Shen L, Cheng AS, Ahmed S, Boumber Y, Charo C et al. (2008). Gene silencing in cancer by histone H3 lysine 27 trimethylation independent of promoter DNA methylation. Nat Genet 40: 741–750.
Martinez-Garcia E, Licht JD . (2010). Deregulation of H3K27 methylation in cancer. Nat Genet 42: 100–101.
Min J, Zaslavsky A, Fedele G, McLaughlin SK, Reczek EE, De Raedt T et al. (2010). An oncogene-tumor suppressor cascade drives metastatic prostate cancer by coordinately activating Ras and nuclear factor-kappaB. Nat Med 16: 286–294.
Mitra RS, Goto M, Lee JS, Maldonado D, Taylor JM, Pan Q et al. (2008). Rap1GAP promotes invasion via induction of matrix metalloproteinase 9 secretion, which is associated with poor survival in low N-stage squamous cell carcinoma. Cancer Res 68: 3959–3969.
Mitra RS, Zhang Z, Henson BS, Kurnit DM, Carey TE, D'Silva NJ . (2003). Rap1A and rap1B ras-family proteins are prominently expressed in the nucleus of squamous carcinomas: nuclear translocation of GTP-bound active form. Oncogene 22: 6243–6256.
Morin RD, Johnson NA, Severson TM, Mungall AJ, An J, Goya R et al. (2010). Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin. Nat Genet 42: 181–185.
Nikoloski G, Langemeijer SM, Kuiper RP, Knops R, Massop M, Tonnissen ER et al. (2010). Somatic mutations of the histone methyltransferase gene EZH2 in myelodysplastic syndromes. Nat Genet 42: 665–667.
Price LS, Hajdo-Milasinovic A, Zhao J, Zwartkruis FJ, Collard JG, Bos JL . (2004). Rap1 regulates E-cadherin-mediated cell-cell adhesion. J Biol Chem 279: 35127–35132.
Reedquist KA, Ross E, Koop EA, Wolthuis RM, Zwartkruis FJ, van Kooyk Y et al. (2000). The small GTPase, Rap1, mediates CD31-induced integrin adhesion. J Cell Biol 148: 1151–1158.
Rubinfeld B, Munemitsu S, Clark R, Conroy L, Watt K, Crosier WJ et al. (1991). Molecular cloning of a GTPase activating protein specific for the Krev-1 protein p21rap1. Cell 65: 1033–1042.
Simon JA, Lange CA . (2008). Roles of the EZH2 histone methyltransferase in cancer epigenetics. Mutat Res 647: 21–29.
Stork PJ, Dillon TJ . (2005). Multiple roles of Rap1 in hematopoietic cells: complementary versus antagonistic functions. Blood 106: 2952–2961.
Su L, Hattori M, Moriyama M, Murata N, Harazaki M, Kaibuchi K et al. (2003). AF-6 controls integrin-mediated cell adhesion by regulating Rap1 activation through the specific recruitment of Rap1GTP and SPA-1. J Biol Chem 278: 15232–15238.
Takai Y, Sasaki T, Matozaki T . (2001). Small GTP-binding proteins. Physiol Rev 81: 153–208.
Thomas GJ, Speight PM . (2001). Cell adhesion molecules and oral cancer. Crit Rev Oral Biol Med 12: 479–498.
Todd R, Donoff RB, Wong DT . (1997). The molecular biology of oral carcinogenesis: toward a tumor progression model. J Oral Maxillofac Surg 55: 613–623; discussion 623–615.
Tsygankova OM, Ma C, Tang W, Korch C, Feldman MD, Lv Y et al. (2010). Downregulation of Rap1GAP in human tumor cells alters cell/matrix and cell/cell adhesion. Mol Cell Biol 30: 3262–3274.
Tsygankova OM, Prendergast GV, Puttaswamy K, Wang Y, Feldman MD, Wang H et al. (2007). Downregulation of Rap1GAP contributes to Ras transformation. Mol Cell Biol 27: 6647–6658.
Varambally S, Cao Q, Mani RS, Shankar S, Wang X, Ateeq B et al. (2008). Genomic loss of microRNA-101 leads to overexpression of histone methyltransferase EZH2 in cancer. Science 322: 1695–1699.
Varambally S, Dhanasekaran SM, Zhou M, Barrette TR, Kumar-Sinha C, Sanda MG et al. (2002). The polycomb group protein EZH2 is involved in progression of prostate cancer. Nature 419: 624–629.
Vire E, Brenner C, Deplus R, Blanchon L, Fraga M, Didelot C et al. (2006). The polycomb group protein EZH2 directly controls DNA methylation. Nature 439: 871–874.
Yang Y, Li X, Yang Q, Wang X, Zhou Y, Jiang T et al. (2010). The role of microRNA in human lung squamous cell carcinoma. Cancer Genet Cytogenet 200: 127–133.
Yarbrough WG, Slebos RJ, Liebler D . (2006). Proteomics: clinical applications for head and neck squamous cell carcinoma. Head Neck 28: 549–558.
Yu J, Cao Q, Mehra R, Laxman B, Tomlins SA, Creighton CJ et al. (2007). Integrative genomics analysis reveals silencing of beta-adrenergic signaling by polycomb in prostate cancer. Cancer Cell 12: 419–431.
Yu J, Mani RS, Cao Q, Brenner CJ, Cao X, Wang X et al. (2010). An integrated network of androgen receptor, polycomb, and TMPRSS2-ERG gene fusions in prostate cancer progression. Cancer Cell 17: 443–454.
Zhang L, Chenwei L, Mahmood R, van Golen K, Greenson J, Li G et al. (2006a). Identification of a putative tumor suppressor gene Rap1GAP in pancreatic cancer. Cancer Res 66: 898–906.
Zhang Z, Mitra RS, Henson BS, Datta NS, McCauley LK, Kumar P et al. (2006b). Rap1GAP inhibits tumor growth in oropharyngeal squamous cell carcinoma. Am J Pathol 168: 585–596.
Zheng H, Gao L, Feng Y, Yuan L, Zhao H, Cornelius LA . (2009). Down-regulation of Rap1GAP via promoter hypermethylation promotes melanoma cell proliferation, survival, and migration. Cancer Res 69: 449–457.
Zuo H, Gandhi M, Edreira MM, Hochbaum D, Nimgaonkar VL, Zhang P et al. (2010). Downregulation of Rap1GAP through epigenetic silencing and loss of heterozygosity promotes invasion and progression of thyroid tumors. Cancer Res 70: 1389–1397.
Acknowledgements
We thank Dr T Giardano for tissue, and Dr Khalid Suleman and Vineeta Sharma for technical assistance for the FISH and methylation studies, respectively. R-SM is the recipient of StewartRahr–PCF Young Investigator Award from the Prostate Cancer Foundation. This work was supported by NIDCR DE16920-01, DE018512-01 grants and funding from University of Michigan Head and Neck Specialized Program of Research Excellence (SPORE) (NJD). This research was made possible, in part, by the use of the Cancer Center Flow Cytometry Core, which was supported (in part) by the National Institutes of Health through the University of Michigan's Cancer Center Support Grant (5 P30 CA46592).
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Banerjee, R., Mani, RS., Russo, N. et al. The tumor suppressor gene rap1GAP is silenced by miR-101-mediated EZH2 overexpression in invasive squamous cell carcinoma. Oncogene 30, 4339–4349 (2011). https://doi.org/10.1038/onc.2011.141
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DOI: https://doi.org/10.1038/onc.2011.141
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