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Tumor Biology

, Volume 35, Issue 12, pp 11819–11827 | Cite as

Epigenetic silencing of methyl-CpG-binding protein 2 gene affects proliferation, invasion, migration, and apoptosis of human osteosarcoma cells

  • Gang Meng
  • YangFan Lv
  • Huanzi Dai
  • Xi Zhang
  • Qiao-Nan Guo
Research Article

Abstract

Methyl-CpG-binding protein 2 (MeCP2) is a DNA methylation-related gene of the methyl-CpG-binding protein family. Here, we investigated the epigenetic function of the MeCP2 in SaOS2 and U2OS cell lines, and explored the antitumor effects of the gene silencing for osteosarcoma. In this study, chromatin immunoprecipitation assay was used to detect MeCP2 binding activity with TSSC3 gene. RT-PCR and western blot assay were used to analyze the MeCP2 expression in osteosarcoma cell lines after transfection with LV-MECP2-RNAi. Transwell invasion and migration assays were used to detect the cell invasion and migration. The cell apoptosis was examined by using the flow cytometry assay. The tumor size was also assessed to determine the therapeutic effects of gene silencing. The results indicated that MeCP2 indicated the highest combining power with TSSC3 gene. LV-MECP2-RNAi could decrease MeCP2 level in tumor cells compared with the untreated cells (P < 0.05). LV-MECP2-RNAi inhibited the U2OS and SaOS2 cells invasion and migration compared with the control cells (P < 0.05). LV-MECP2-RNAi triggered the U2OS and SaOS2 cell apoptosis, and inhibited the cell proliferation significantly compared with the control cells (P < 0.05). The gene silencing of RNAi could also decreased the tumor size significantly compared with untreated cells (P < 0.05). In conclusion, silencing the MeCP2 gene could block the MeCP2 expression and inhibit the tumor cell migration, invasion, and proliferation, and decreases the tumor size by inducing the apoptosis of the tumor cells.

Keywords

Methyl-CpG-binding protein 2 Osteosarcoma Apoptosis Proliferation 

Notes

Acknowledgments

This research was supported by a grant from the National Natural Science Foundation of China (No. 81001198 and 81372864).

Conflicts of interest

None

References

  1. 1.
    Huang J, Gao K, Lin J, Wang Q. MicroRNA-100 inhibits osteosarcoma cells proliferation by targeting Cyr61. Tumor Biol. 2014;35(2):1095–100.CrossRefGoogle Scholar
  2. 2.
    Ando K, Heymann MF, Stresing V, Mori K, Redini F, Heymann D. Current therapeutic strategies and novel approaches in osteosarcoma. Cancer (Basel). 2013;5(2):591–616.CrossRefGoogle Scholar
  3. 3.
    Meyers PA, Gorlick R. Osteosarcoma. Pediatr Clin North Am. 1997;44(4):973–89.PubMedCrossRefGoogle Scholar
  4. 4.
    Miao J, Wu S, Peng Z, Tania M, Zhang C. MicroRNAs in osteosarcoma: diagnostic and therapeutic aspects. Tumor Biol. 2013;34(4):2093–8.CrossRefGoogle Scholar
  5. 5.
    Stiller CA. International patterns of cancer incidence in adolescents. Cancer Treat Rev. 2007;33(7):631–45.PubMedCrossRefGoogle Scholar
  6. 6.
    Coventry MB, Mahlin DC. Osteogenic sarcoma: a critical analysis of 430 cases. J Bone Joint Surg Am. 1957;39(4):741–57.PubMedGoogle Scholar
  7. 7.
    Kager L, Zoubek A, Potschger U, Kastner U, Flege S, Kempf-Bielack B, et al. Primary metastatic osteosarcoma: presentation and outcome of patients treated on neoadjuvant cooperative osteosarcoma study group protocols. J Clin Oncol. 2003;21(10):2011–8.PubMedCrossRefGoogle Scholar
  8. 8.
    Jones PA. DNA methylation and cancer. Oncogene. 2002;2:5358–60.CrossRefGoogle Scholar
  9. 9.
    Siedlecki P, Zielenkiewicz P. Mammalian DNA methyltransferases. Acta Biochim Pol. 2006;53(2):245–56.PubMedGoogle Scholar
  10. 10.
    Lyko F, Brown R. DNA methyltransferase inhibitors and the development of epigenetic cancer therapies. J Natl Cancer Inst. 2005;97(20):1498–506.PubMedCrossRefGoogle Scholar
  11. 11.
    Mirza S, Sharma G, Parshad R, Gupta SD, Pandya P, Ralhan R. Expression of DNA methyltransferases in breast cancer patients and to analyze the effect of natural compounds on DNA methyltransferases and associated proteins. J Breast Cancer. 2013;16(1):23–31.PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Yi JM, Dhir M, Guzzetta AA, Lacobuzio-Donahue CA, Heo K, Yang KM, et al. DNA methylation biomarker candidates for early detection of colon cancer. Tumor Biol. 2013;33(2):363–72.CrossRefGoogle Scholar
  13. 13.
    Muotri AR, Marchetto MC, Coufal NG, Oefner R, Yeo G, Nakashima K, et al. L1 retrotransposition in neurons is modulated by MeCP2. Nature. 2011;468(7322):443–6.CrossRefGoogle Scholar
  14. 14.
    Konduri SD, Srivenugopal KS, Yanamadra N, Dinh DH, Olivero WC. Promoter methylation and silencing of the tissue factor pathway inhibitor-2 (TFPI-2), a gene encoding an inhibitor of matrix metalloproteinases in human glioma cells. Oncogene. 2003;22(29):4509–16.PubMedCrossRefGoogle Scholar
  15. 15.
    Murphy DM, Buckley PG, Das S, Watters KM, Bryan K, Stallings RL. Co-localization of the oncogenic transcription factor MYCN and the DNA methyl binding protein MeCP2 at genomic sites in neuroblastoma. PloS One. 6(6):e21436.Google Scholar
  16. 16.
    Wang X, Shi Q, Xu K, Gao C, Chen C, Li XL, et al. Familial CJD associated PrP mutants within transmembrane region induced Ctm-PrP retention in ER and triggered apoptosis by ER stress in SH-SY5Y cells. PLoS One. 2011;6(1):e14602.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Reynolds A, Leake D, Boese Q, Scaringe S, Marshall WS, Khvorova A. Rational siRNA design for RNA interference. Nat Biotechnol. 2004;22(3):326–30.PubMedCrossRefGoogle Scholar
  18. 18.
    Squillaro T, Alessio N, Cipollaro M, Renieri A, Giordano A, Galderisi U. Partial silencing of methyl cytosine protein binding 2 (MECP2) in mesenchymal stem cells induces senescence with an increase in damaged DNA. FASEB J. 2010;24(5):1593–603.PubMedCrossRefGoogle Scholar
  19. 19.
    Esteller M. Epigenetics in cancer. N Engl J Med. 2008;358:1148–59.PubMedCrossRefGoogle Scholar
  20. 20.
    Li Y, Tollefsbol TO. Impact on DNA methylation in cancer prevention and therapy by bioactive dietary components. Curr Med Chem. 2010;17(20):2141–51.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Prokhortchouk E, Hendrich B. Methyl-CpG binding proteins and cancer: are MeCpGs more important than MBDs? Oncogene. 2002;21(35):5394–9.PubMedCrossRefGoogle Scholar
  22. 22.
    Pontes TB, Chen ES, Gigek CO, Calcagno DQ, Wisnieski F, Leal MF, et al. Reduced mRNA expression levels of MBD2 and MBD3 in gastric carcinogenesis. Tumor Biol. 2014;35(4):3447–53.CrossRefGoogle Scholar
  23. 23.
    Garinis GA, Patrinos GP, Spanakis NE, Menounos PG. DNA hypermethylation: when tumour suppressor genes go silent. Hum Genet. 2002;111(2):115–27.PubMedCrossRefGoogle Scholar
  24. 24.
    Leoh LS, van Heertum B, De Rijck J, Filippova M, Rios-Colon L, Basu A, et al. The stress oncoprotein LEDGF/p75 interacts with the methyl CpG binding protein MeCP2 and influences its transcriptional activity. Mol Cancer Res. 2012;10(3):378–91.PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Takao T, Asanoma K, Tsunematsu R, Kato K, Wake N. The maternally expressed gene Tssc3 regulates the expression of MASH2 transcription factor in mouse trophoblast stem cells through the AKT-Sp1 signaling pathway. J Biol Chem. 2012;287(51):42685–94.PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Sanchez-Carbayo M. Hypermethylation in bladder cancer: biological pathways and translational applications. Tumor Biol. 2012;33(2):347–61.CrossRefGoogle Scholar
  27. 27.
    Peddada S, Yasui DH, LaSalle JM. Inhibitors of differentiation (ID1, ID2, ID3 and ID4) genes are neuronal targets of MeCP2 that are elevated in Rett syndrome. Hum Mol Genet. 2006;15(12):2003–14.PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Darwanto A, Kitazawa R, Maeda S, Kitazawa S. MeCP2 and promoter methylation cooperatively regulate E-cadherin gene expression in colorectal carcinoma. Cancer Sci. 2003;94(5):442–7.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Gang Meng
    • 1
  • YangFan Lv
    • 1
  • Huanzi Dai
    • 2
  • Xi Zhang
    • 1
  • Qiao-Nan Guo
    • 1
  1. 1.Department of Pathology, Xinqiao HospitalThird Military Medical UniversityChongqingChina
  2. 2.Department of Nephrology, Daping HospitalThird Military Medical UniversityChongqingChina

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