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Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen

Oncogene volume 30, pages 3875–3886 (2011)Cite this article

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Abstract

The introduction of the Simian virus 40 (SV40) early region, the telomerase catalytic subunit (hTERT) and an oncogenic allele of H-Ras directly transforms primary human cells. SV40 small T antigen (ST), which forms a complex with protein phosphatase 2A (PP2A) and inhibits PP2A activity, is believed to have a critical role in the malignant transformation of human cells. Recent evidence has shown that aberrant microRNA (miRNA) expression patterns are correlated with cancer development. Here, we identified miR-27a as a differentially expressed miRNA in SV40 ST-expressing cells. miR-27a is upregulated in SV40 ST-transformed human bronchial epithelial cells (HBERST). Suppression of miR-27a expression in HBERST cells or lung cancer cell lines (NCI-H226 and SK-MES-1) that exhibited high levels of miR-27a expression lead to cell growth arrested in the G0–G1 phase. In addition, suppression of miR-27a in HBERST cells attenuated the capacity of such cells to grow in an anchorage-independent manner. We also found that suppression of the PP2A B56γ expression resulted in upregulation of miR-27a similar to that achieved by the introduction of ST, indicating that dysregulation of miR-27a expression in ST-expressing cells was mediated by the ST–PP2A interaction. Moreover, we discovered that Fbxw7 gene encoding F-box/WD repeat-containing protein 7 was a potential miR-27a target validated by dual-luciferase reporter system analysis. The inverse correlation between miR-27a expression levels and Fbxw7 protein expression was further confirmed in both cell models and human tumor samples. Fbxw7 regulates cell-cycle progression through the ubiquitin-dependent proteolysis of a set of substrates, including c-Myc, c-Jun, cyclin E1 and Notch 1. Thus, promotion of cell growth arising from the suppression of Fbxw7 by miR-27a overexpression might be responsible for the viral oncoprotein ST-induced malignant transformation. These observations demonstrate that miR-27a functions as an oncogene in human tumorigenesis.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

ASO:

antisense oligonucleotide

Fbxw7:

F-box/WD repeat-containing protein 7

HBE cells:

human bronchial epithelial cells

hTERT :

human telomerase catalytic subunit

NC:

negative control

OA:

okadaic acid

PP2A:

protein phosphatase 2A

qRT–PCR:

quantitative real-time PCR

shB56γ:

small-hairpin RNA interference against the B56γ subunit

ST:

SV40 small T antigen

HBER cells:

cells expressing hTERT, the SV40 large T antigen (LT) and an oncogenic allele of H-Ras

References

  • Akhoondi S, Sun D, von der Lehr N, Apostolidou S, Klotz K, Maljukova A et al. (2007). FBXW7/hCDC4 is a general tumor suppressor in human cancer. Cancer Res 67: 9006–9012.

    Article  CAS  PubMed  Google Scholar 

  • Ali SH, DeCaprio JA . (2001). Cellular transformation by SV40 large T antigen: interaction with host proteins. Semin Cancer Biol 11: 15–23.

    Article  CAS  PubMed  Google Scholar 

  • Ambros V . (2004). The functions of animal microRNAs. Nature 431: 350–355.

    Article  CAS  PubMed  Google Scholar 

  • Arnold HK, Sears RC . (2006). Protein phosphatase 2A regulatory subunit B56alpha associates with c-myc and negatively regulates c-myc accumulation. Mol Cell Biol 26: 2832–2844.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arroyo JD, Hahn WC . (2005). Involvement of PP2A in viral and cellular transformation. Oncogene 24: 7746–7755.

    Article  CAS  PubMed  Google Scholar 

  • Bartel DP . (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116: 281–297.

    Article  CAS  PubMed  Google Scholar 

  • Cegielska A, Shaffer S, Derua R, Goris J, Virshup DM . (1994). Different oligomeric forms of protein phosphatase 2A activate and inhibit simian virus 40 DNA replication. Mol Cell Biol 14: 4616–4623.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen W, Hahn WC . (2003). SV40 early region oncoproteins and human cell transformation. Histol Histopathol 18: 541–550.

    CAS  PubMed  Google Scholar 

  • Chen W, Possemato R, Campbell KT, Plattner CA, Pallas DC, Hahn WC . (2004). Identification of specific PP2A complexes involved in human cell transformation. Cancer Cell 5: 127–136.

    Article  CAS  PubMed  Google Scholar 

  • Choi YW, Lee IC, Ross SR . (1988). Requirement for the simian virus 40 small tumor antigen in tumorigenesis in transgenic mice. Mol Cell Biol 8: 3382–3390.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cohen P . (1989). The structure and regulation of protein phosphatases. Annu Rev Biochem 58: 453–508.

    Article  CAS  PubMed  Google Scholar 

  • Connolly E, Melegari M, Landgraf P, Tchaikovskaya T, Tennant BC, Slagle BL et al. (2008). Elevated expression of the miR-17-92 polycistron and miR-21 in hepadnavirus-associated hepatocellular carcinoma contributes to the malignant phenotype. Am J Pathol 173: 856–864.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Crawford LV, Cole CN, Smith AE, Paucha E, Tegtmeyer P, Rundell K et al. (1978). Organization and expression of early genes of simian virus 40. Proc Natl Acad Sci USA 75: 117–121.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Crusio KM, King B, Reavie LB, Aifantis I . (2010). The ubiquitous nature of cancer: the role of the SCF(Fbw7) complex in development and transformation. Oncogene 29: 4865–4873.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eichhorn PJ, Creyghton MP, Bernards R . (2009). Protein phosphatase 2A regulatory subunits and cancer. Biochim Biophys Acta 1795: 1–15.

    CAS  PubMed  Google Scholar 

  • Engels BM, Hutvagner G . (2006). Principles and effects of microRNA-mediated post-transcriptional gene regulation. Oncogene 25: 6163–6169.

    Article  CAS  PubMed  Google Scholar 

  • Esquela-Kerscher A, Slack FJ . (2006). Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer 6: 259–269.

    Article  CAS  PubMed  Google Scholar 

  • Ferrigno P, Langan TA, Cohen P . (1993). Protein phosphatase 2A1 is the major enzyme in vertebrate cell extracts that dephosphorylates several physiological substrates for cyclin-dependent protein kinases. Mol Biol Cell 4: 669–677.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Godshalk SE, Bhaduri-McIntosh S, Slack FJ . (2008). Epstein-Barr virus-mediated dysregulation of human microRNA expression. Cell Cycle 7: 3595–3600.

    Article  CAS  PubMed  Google Scholar 

  • Grochola LF, Vazquez A, Bond EE, Wurl P, Taubert H, Muller TH et al. (2009). Recent natural selection identifies a genetic variant in a regulatory subunit of protein phosphatase 2A that associates with altered cancer risk and survival. Clin Cancer Res 15: 6301–6308.

    Article  CAS  PubMed  Google Scholar 

  • Guo H, Ingolia NT, Weissman JS, Bartel DP . (2010). Mammalian microRNAs predominantly act to decrease target mRNA levels. Nature 466: 835–840.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guttilla IK, White BA . (2009). Coordinate regulation of FOXO1 by miR-27a, miR-96, and miR-182 in breast cancer cells. J Biol Chem 284: 23204–23216.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hahn WC, Counter CM, Lundberg AS, Beijersbergen RL, Brooks MW, Weinberg RA . (1999). Creation of human tumour cells with defined genetic elements. Nature 400: 464–468.

    Article  CAS  PubMed  Google Scholar 

  • Hahn WC, Dessain SK, Brooks MW, King JE, Elenbaas B, Sabatini DM et al. (2002). Enumeration of the simian virus 40 early region elements necessary for human cell transformation. Mol Cell Biol 22: 2111–2123.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hirakawa T, Ruley HE . (1988). Rescue of cells from ras oncogene-induced growth arrest by a second, complementing, oncogene. Proc Natl Acad Sci USA 85: 1519–1523.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Huang S, He X, Ding J, Liang L, Zhao Y, Zhang Z et al. (2008). Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells. Int J Cancer 123: 972–978.

    Article  CAS  PubMed  Google Scholar 

  • Janssens V, Goris J . (2001). Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochem J 353: 417–439.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Khalili K, Brady J, Khoury G . (1987). Translational regulation of SV40 early mRNA defines a new viral protein. Cell 48: 639–645.

    Article  CAS  PubMed  Google Scholar 

  • Knuutila S, Aalto Y, Autio K, Bjorkqvist AM, El-Rifai W, Hemmer S et al. (1999). DNA copy number losses in human neoplasms. Am J Pathol 155: 683–694.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Koepp DM, Schaefer LK, Ye X, Keyomarsi K, Chu C, Harper JW et al. (2001). Phosphorylation-dependent ubiquitination of cyclin E by the SCFFbw7 ubiquitin ligase. Science 294: 173–177.

    Article  CAS  PubMed  Google Scholar 

  • Lai EC . (2004). Predicting and validating microRNA targets. Genome Biol 5: 115.

    Article  PubMed  PubMed Central  Google Scholar 

  • Lewis BP, Burge CB, Bartel DP . (2005). Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120: 15–20.

    Article  CAS  PubMed  Google Scholar 

  • Liu T, Tang H, Lang Y, Liu M, Li X . (2009). MicroRNA-27a functions as an oncogene in gastric adenocarcinoma by targeting prohibitin. Cancer Lett 273: 233–242.

    Article  CAS  PubMed  Google Scholar 

  • Ma Y, Yu S, Zhao W, Lu Z, Chen J . (2010). miR-27a regulates the growth, colony formation and migration of pancreatic cancer cells by targeting Sprouty2. Cancer Lett 298: 150–158.

    Article  CAS  PubMed  Google Scholar 

  • Mao JH, Perez-Losada J, Wu D, Delrosario R, Tsunematsu R, Nakayama KI et al. (2004). Fbxw7/Cdc4 is a p53-dependent, haploinsufficient tumour suppressor gene. Nature 432: 775–779.

    Article  CAS  PubMed  Google Scholar 

  • Margolis SS, Perry JA, Forester CM, Nutt LK, Guo Y, Jardim MJ et al. (2006). Role for the PP2A/B56delta phosphatase in regulating 14-3-3 release from Cdc25 to control mitosis. Cell 127: 759–773.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mertens-Talcott SU, Chintharlapalli S, Li X, Safe S . (2007). The oncogenic microRNA-27a targets genes that regulate specificity protein transcription factors and the G2-M checkpoint in MDA-MB-231 breast cancer cells. Cancer Res 67: 11001–11011.

    Article  CAS  PubMed  Google Scholar 

  • Michalovitz D, Fischer-Fantuzzi L, Vesco C, Pipas JM, Oren M . (1987). Activated Ha-ras can cooperate with defective simian virus 40 in the transformation of nonestablished rat embryo fibroblasts. J Virol 61: 2648–2654.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Moreno CS, Ramachandran S, Ashby DG, Laycock N, Plattner CA, Chen W et al. (2004). Signaling and transcriptional changes critical for transformation of human cells by simian virus 40 small tumor antigen or protein phosphatase 2A B56gamma knockdown. Cancer Res 64: 6978–6988.

    Article  CAS  PubMed  Google Scholar 

  • Mumby M . (1995). Regulation by tumour antigens defines a role for PP2A in signal transduction. Semin Cancer Biol 6: 229–237.

    Article  CAS  PubMed  Google Scholar 

  • Pallas DC, Shahrik LK, Martin BL, Jaspers S, Miller TB, Brautigan DL et al. (1990). Polyoma small and middle T antigens and SV40 small t antigen form stable complexes with protein phosphatase 2A. Cell 60: 167–176.

    Article  CAS  PubMed  Google Scholar 

  • Pang Y, Li W, Ma R, Ji W, Wang Q, Li D et al. (2008). Development of human cell models for assessing the carcinogenic potential of chemicals. Toxicol Appl Pharmacol 232: 478–486.

    Article  CAS  PubMed  Google Scholar 

  • Perez-Losada J, Mao JH, Balmain A . (2005). Control of genomic instability and epithelial tumor development by the p53-Fbxw7/Cdc4 pathway. Cancer Res 65: 6488–6492.

    Article  CAS  PubMed  Google Scholar 

  • Pichler K, Schneider G, Grassmann R . (2008). MicroRNA miR-146a and further oncogenesis-related cellular microRNAs are dysregulated in HTLV-1-transformed T lymphocytes. Retrovirology 5: 100.

    Article  PubMed  PubMed Central  Google Scholar 

  • Rhodes DR, Kalyana-Sundaram S, Mahavisno V, Varambally R, Yu J, Briggs BB et al. (2007). Oncomine 3.0: genes, pathways, and networks in a collection of 18 000 cancer gene expression profiles. Neoplasia 9: 166–180.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rundell K, Parakati R . (2001). The role of the SV40 ST antigen in cell growth promotion and transformation. Semin Cancer Biol 11: 5–13.

    Article  CAS  PubMed  Google Scholar 

  • Sablina AA, Hahn WC . (2008). SV40 small T antigen and PP2A phosphatase in cell transformation. Cancer Metastasis Rev 27: 137–146.

    Article  CAS  PubMed  Google Scholar 

  • Shenk TE, Carbon J, Berg P . (1976). Construction and analysis of viable deletion mutants of simian virus 40. J Virol 18: 664–671.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Shouse GP, Nobumori Y, Liu X . (2010). A B56gamma mutation in lung cancer disrupts the p53-dependent tumor-suppressor function of protein phosphatase 2A. Oncogene 29: 3933–3941.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sleigh MJ, Topp WC, Hanich R, Sambrook JF . (1978). Mutants of SV40 with an altered small t protein are reduced in their ability to transform cells. Cell 14: 79–88.

    Article  CAS  PubMed  Google Scholar 

  • Sontag E, Fedorov S, Kamibayashi C, Robbins D, Cobb M, Mumby M . (1993). The interaction of SV40 small tumor antigen with protein phosphatase 2A stimulates the map kinase pathway and induces cell proliferation. Cell 75: 887–897.

    Article  CAS  PubMed  Google Scholar 

  • Sontag JM, Sontag E . (2006). Regulation of cell adhesion by PP2A and SV40 small tumor antigen: an important link to cell transformation. Cell Mol Life Sci 63: 2979–2991.

    Article  CAS  PubMed  Google Scholar 

  • Tan Y, Sangfelt O, Spruck C . (2008). The Fbxw7/hCdc4 tumor suppressor in human cancer. Cancer Lett 271: 1–12.

    Article  CAS  PubMed  Google Scholar 

  • Tomita M, Tanaka Y, Mori N . (2009). MicroRNA miR-146a is induced by HTLV-1 tax and increases the growth of HTLV-1-infected T-cells. Int J Cancer (in press).

  • Tsunematsu R, Nakayama K, Oike Y, Nishiyama M, Ishida N, Hatakeyama S et al. (2004). Mouse Fbw7/Sel-10/Cdc4 is required for notch degradation during vascular development. J Biol Chem 279: 9417–9423.

    Article  CAS  PubMed  Google Scholar 

  • Wei W, Jin J, Schlisio S, Harper JW, Kaelin Jr WG . (2005). The v-Jun point mutation allows c-Jun to escape GSK3-dependent recognition and destruction by the Fbw7 ubiquitin ligase. Cancer Cell 8: 25–33.

    Article  CAS  PubMed  Google Scholar 

  • Welcker M, Orian A, Jin J, Grim JE, Harper JW, Eisenman RN et al. (2004). The Fbw7 tumor suppressor regulates glycogen synthase kinase 3 phosphorylation-dependent c-Myc protein degradation. Proc Natl Acad Sci USA 101: 9085–9090.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yang SI, Lickteig RL, Estes R, Rundell K, Walter G, Mumby MC . (1991). Control of protein phosphatase 2A by simian virus 40 small-t antigen. Mol Cell Biol 11: 1988–1995.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yu J, Boyapati A, Rundell K . (2001). Critical role for SV40 small-t antigen in human cell transformation. Virology 290: 192–198.

    Article  CAS  PubMed  Google Scholar 

  • Zeng C, Wang R, Li D, Lin XJ, Wei QK, Yuan Y et al. (2010). A novel GSK-3 beta-C/EBP alpha-miR-122-insulin-like growth factor 1 receptor regulatory circuitry in human hepatocellular carcinoma. Hepatology 52: 1702–1712.

    Article  CAS  PubMed  Google Scholar 

  • Zerrahn J, Knippschild U, Winkler T, Deppert W . (1993). Independent expression of the transforming amino-terminal domain of SV40 large I antigen from an alternatively spliced third SV40 early mRNA. EMBO J 12: 4739–4746.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao Y, Ransom JF, Li A, Vedantham V, von Drehle M, Muth AN et al. (2007). Dysregulation of cardiogenesis, cardiac conduction, and cell cycle in mice lacking miRNA-1-2. Cell 129: 303–317.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by program of Distinguished Young Scholar of NSFC (30925029, 30925036), a Key NSFC Program (30630055) and NSFC (30800930, 30771832, 30901211), National Key Basic Research and Development Program (2010CB912803), National High Technology Research and Development Key Program of China (2008AA062504), Ministry of Health of China (200902006), the Fundamental Research Funds for the Central Universities (10ykjc05 and 10lgzd10), Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme GDUPS (2010).

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  1. Q Wang and D-C Li: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Toxicology, Guangdong Provincial Key Laboratory of Food, Nutrition and Health, School of Public Health, Sun Yat-sen University, Guangzhou, China

    Q Wang, D-C Li, Z-F Li, C-X Liu, Y-M Xiao, B Zhang, L-P Chen, X-M Xing, S-F Tang, Y-C Lin, Y-D Lai, P Yang, J-L Zeng, Q Xiao, X-W Zeng, Z-N Lin, Z-X Zhuang, S-M Zhuang & W Chen

  2. Department of Thoracic Surgery, Sun Yat-sen University Cancer Center, Guangzhou, China

    X-D Li

  3. Department of Thoracic Surgery, The Affiliated Cancer Hospital of Guangzhou Medical University, Guangzhou, China

    J Zhao

  4. Department of Toxicology, Shenzhen Center for Disease Control and Prevention, Shenzhen, China

    Z-X Zhuang

  5. Sun Yat-sen University, School of Life Sciences Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory of Biocontrol, Guangzhou, China

    S-M Zhuang

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Wang, Q., Li, DC., Li, ZF. et al. Upregulation of miR-27a contributes to the malignant transformation of human bronchial epithelial cells induced by SV40 small T antigen. Oncogene 30, 3875–3886 (2011). https://doi.org/10.1038/onc.2011.103

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