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MicroRNA-148a is silenced by hypermethylation and interacts with DNA methyltransferase 1 in gastric cancer

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Abstract

Studies have shown that microRNA-148a (miR-148a) was proved to be silenced while DNA methyltransferase 1 (DNMT1) was over-expressed in gastric cancer. But the mechanism of aberrant expression of miR-148a and DNMT1 and their relationships in gastric cancer are still unknown. The aims of this study were to investigate the expression profile of miR-148a and DNMT1 and reveal whether they have any relationships. We used reverse-transcriptase quantitative real-time PCR, methylation-specific PCR and Western blot to measure the level of miR-148a expression, DNA methylation level and DNMT1 expression, respectively. Gastric cancer cells were transfected with plasmid or siRNA or treated with 5-aza-2′-deoxycytidine. Cell proliferation and apoptosis were detected by cell counting and flow cytometric analysis. In this study, we demonstrated that gastric cancer tissues and cell lines displayed a consistent down-regulation of miR-148a and hypermethylation of promoter region. DNMT1 was over-expressed in primary tumors and cell lines, while knockdown of DNMT1 using siRNA could decrease methylation level of miR-148a promoter and restore its expression. Furthermore, ectopic over-expression of miR-148a in cancer cell lines caused reduction in DNMT1 expression and inhibited cell proliferation, but no obvious change was found in apoptosis rate. These results suggest that miR-148a is inactivated by DNA hypermethylation of promoter region in gastric cancer, which is mediated through DNMT1 over-expression. Additionally, the silence of miR-148a reduces its suppression to DNMT1 in gastric cancer, and this may in turn result in over-expression of DNMT1 and promote DNA hypermethylation.

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References

  1. Parkin DM, Bray F, Ferlay J, et al. Global cancer statistics, 2002. CA Cancer J Clin. 2005;55:74–108.

    Article  PubMed  Google Scholar 

  2. Dykxhoorn DM. MicroRNAs and metastasis: little RNAs go a long way. Cancer Res. 2010;70:6401–6.

    Article  PubMed  CAS  Google Scholar 

  3. Ardekani AM, Naeini MM. The Role of microRNAs in human diseases. Avicenna J Med Biotech. 2010;2:161–79.

    CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  5. Ueda T, Volinia S, Okumura H, et al. Relation between microRNA expression and progression and prognosis of gastric cancer: a microRNA expression analysis. Lancet Oncol. 2010;11:136–46.

    Article  PubMed  CAS  Google Scholar 

  6. Tsukamoto Y, Nakada C, Noguchi T, et al. MicroRNA-375 is downregulated in gastric carcinomas and regulates cell survival by targeting PDK1 and 14-3-3zeta. Cancer Res. 2010;70:2339–49.

    Article  PubMed  CAS  Google Scholar 

  7. Nguyen T, Kuo C, Nicholl MB, et al. Downregulation of microRNA-29c is associated with hypermethylation of tumor-related genes and disease outcome in cutaneous melanoma. Epigenetics. 2011;6:388–94.

    Article  PubMed  CAS  Google Scholar 

  8. Datta J, Kutay H, Nasser MW, et al. Methylation mediated silencing of microRNA-1 gene and its role in hepatocellular carcinogenesis. Cancer Res. 2008;68:5049–58.

    Article  PubMed  CAS  Google Scholar 

  9. Braconi C, Huang N, Patel T. MicroRNA-dependent regulation of DNA methyltransferase-1 and tumor suppressor gene expression by interleukin-6 in human malignant cholangiocytes. Hepatology. 2010;51:881–90.

    PubMed  CAS  Google Scholar 

  10. Leonhardt H, Page AW, Weier HU, et al. A targeting sequence directs DNA methyltransferase to sites of DNA replication in mammalian nuclei. Cell. 1992;71:865–73.

    Article  PubMed  CAS  Google Scholar 

  11. Bestor T, Laudano A, Mattaliano R, et al. Cloning and sequencing of a cDNA encoding DNA methyltransferase of mouse cells. The carboxyl-terminal domain of the mammalian enzymes is related to bacterial restriction methyltransferases. J Mol Biol. 1988;203:971–83.

    Article  PubMed  CAS  Google Scholar 

  12. Okano M, Bell DW, Haber DA, et al. DNA methyltransferases Dnmt3a and Dnmt3b are essential for de novo methylation and mammalian development. Cell. 1999;99:247–57.

    Article  PubMed  CAS  Google Scholar 

  13. Chedin F, Lieber MR, Hsieh CL. The DNA methyltransferase-like protein DNMT3L stimulates de novo methylation by Dnmt3a. Proc Natl Acad Sci USA. 2002;99:16916–21.

    Article  PubMed  CAS  Google Scholar 

  14. Ooi SK, Qiu C, Bernstein E, et al. DNMT3L connects unmethylated lysine 4 of histone H3 to de novo methylation of DNA. Nature. 2007;448:714–7.

    Article  PubMed  CAS  Google Scholar 

  15. Garzon R, Liu S, Fabbri M, et al. MicroRNA-29b induces global DNA hypomethylation and tumor suppressor gene reexpression in acute myeloid leukemia by targeting directly DNMT3A and 3B and indirectly DNMT1. Blood. 2009;113:6411–8.

    Article  PubMed  CAS  Google Scholar 

  16. Wang H, Wu J, Meng X, et al. MicroRNA-342 inhibits colorectal cancer cell proliferation and invasion by directly targeting DNA methyltransferase 1. Carcinogenesis. 2011;32:1033–42.

    Article  PubMed  CAS  Google Scholar 

  17. Tsuruta T, Kozaki K, Uesugi A, et al. miR-152 is a tumor suppressor microRNA that is silenced by DNA hypermethylation in endometrial cancer. Cancer Res. 2011;71(20):6450–62.

    Article  PubMed  CAS  Google Scholar 

  18. Huang J, Wang Y, Guo Y, et al. Down-regulated microRNA-152 induces aberrant DNA methylation in hepatitis B virus-related hepatocellular carcinoma by targeting DNA methyltransferase 1. Hepatology. 2010;52:60–70.

    Article  PubMed  CAS  Google Scholar 

  19. Duursma AM, Kedde M, Schrier M, et al. miR-148 targets human DNMT3b protein coding region. RNA. 2008;14:872–7.

    Article  PubMed  CAS  Google Scholar 

  20. Chen Y, Song Y, Wang Z, et al. Altered expression of miR-148a and miR-152 in gastrointestinal cancers and its clinical significance. J Gastrointest Surg. 2010;14:1170–9.

    Article  PubMed  Google Scholar 

  21. Katada T, Ishiguro H, Kuwabara Y, et al. MicroRNA expression profile in undifferentiated gastric cancer. Int J Oncol. 2009;34:537–42.

    PubMed  CAS  Google Scholar 

  22. Tseng CW, Lin CC, Chen CN, et al. Integrative network analysis reveals active microRNAs and their functions in gastric cancer. BMC Syst Biol. 2011;5:99.

    Article  PubMed  Google Scholar 

  23. Hino R, Uozaki H, Murakami N, et al. Activation of DNA methyltransferase 1 by EBV latent membrane protein 2A leads to promoter hypermethylation of PTEN gene in gastric carcinoma. Cancer Res. 2009;69:2766–74.

    Article  PubMed  CAS  Google Scholar 

  24. Mutze K, Langer R, Schumacher F, et al. DNA methyltransferase 1 as a predictive biomarker and potential therapeutic target for chemotherapy in gastric cancer. Eur J Cancer. 2011;47:1817–25.

    Article  PubMed  CAS  Google Scholar 

  25. Hanoun N, Delpu Y, Suriawinata AA, et al. The silencing of microRNA 148a production by DNA hypermethylation is an early event in pancreatic carcinogenesis. Clin Chem. 2010;56:1107–18.

    Article  PubMed  CAS  Google Scholar 

  26. Berdiev BK, Xia J, McLean LA, et al. Acid-sensing ion channels in malignant gliomas. J Biol Chem. 2003;278:15023–34.

    Article  PubMed  CAS  Google Scholar 

  27. Esteller M. CpG island hypermethylation and tumor suppressor genes: a booming present, a brighter future. Oncogene. 2002;21:5427–40.

    Article  PubMed  CAS  Google Scholar 

  28. Lehmann U, Hasemeier B, Christgen M, et al. Epigenetic inactivation of microRNA gene hsa-mir-9-1 in human breast cancer. J Pathol. 2008;214:17–24.

    Article  PubMed  CAS  Google Scholar 

  29. Pan W, Zhu S, Yuan M, et al. MicroRNA-21 and microRNA-148a contribute to DNA hypomethylation in Lupus CD4+T cells by directly and indirectly targeting DNA methyltransferase 1. J Immunol. 2010;184:6773–81.

    Article  PubMed  CAS  Google Scholar 

  30. Braconi C, Kogure T, Valeri N, et al. microRNA-29 can regulate expression of the long non-coding RNA gene MEG3 in hepatocellular cancer. Oncogene. 2011;30:4750–6.

    Article  PubMed  CAS  Google Scholar 

  31. Lim LP, Lau NC, Garrett-Engele P, et al. Microarray analysis shows that some microRNAs down-regulate large numbers of target mRNAs. Nature. 2005;433:769–73.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Key Program for Technology Development Fund of Nanjing Medical University (NJMU04).

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The authors have no conflict of interest to declare.

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Authors and Affiliations

  1. Department of General Surgery, Nanjing Medical University Affiliated Wuxi Second Hospital, No. 68, ZhongShan Road of Wuxi City, Wuxi, 214002, JiangSu Province, People’s Republic of China

    Akao Zhu, Jiazeng Xia, Junbo Zuo, Hong Zhou & Lubin Yao

  2. Department of Gastroenterology, Nanjing Medical University Affiliated Wuxi Second Hospital, Wuxi, 214002, JiangSu Province, People’s Republic of China

    Shimao Jin

  3. Central Laboratory, Nanjing Medical University Affiliated Wuxi Second Hospital, Wuxi, 214002, JiangSu Province, People’s Republic of China

    Hongyu Huang & Zhijun Han

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  1. Akao Zhu
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  2. Jiazeng Xia
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  3. Junbo Zuo
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  4. Shimao Jin
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  5. Hong Zhou
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  7. Hongyu Huang
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Correspondence to Jiazeng Xia.

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Zhu, A., Xia, J., Zuo, J. et al. MicroRNA-148a is silenced by hypermethylation and interacts with DNA methyltransferase 1 in gastric cancer. Med Oncol 29, 2701–2709 (2012). https://doi.org/10.1007/s12032-011-0134-3

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  • DOI: https://doi.org/10.1007/s12032-011-0134-3

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