Tumor Biology

, Volume 36, Issue 4, pp 3009–3015 | Cite as

Preliminary screening of differentially expressed genes involved in methyl-CpG-binding protein 2 gene-mediated proliferation in human osteosarcoma cells

  • Gang Meng
  • Yi Li
  • YangFan lv
  • Huanzi Dai
  • Xi Zhang
  • Qiao-Nan Guo
Research Article

Abstract

Methyl-CpG-binding protein 2 (MeCP2) is essential in human brain development and has been linked to several cancer types and neuro-developmental disorders. This study aims to screen the MeCP2 related differentially expressed genes and discover the therapeutic targets for osteosarcoma. CCK8 assay was used to detect the proliferation and SaOS2 and U2OS cells. Apoptosis of cells was detected by flow cytometry analysis that monitored Annexin V-APC/7-DD binding and 7-ADD uptake simultaneously. Denaturing formaldehyde agarose gel electrophoresis was employed to examine the quality of total RNA 18S and 28S units. Gene chip technique was utilized to discover the differentially expressed genes correlated with MeCP2 gene. Differential gene screening criteria were used to screen the changed genes. The gene up-regulation or down-regulation more than 1.5 times was regarded as significant differential expression genes. The CCK8 results indicated that the cell proliferation of MeCP2 silencing cells (LV-MeCP2-RNAi) was significantly decreased compared to non-silenced cells (LV-MeCP2-RNAi-CN) (P < 0.05). MeCP2 silencing could also induce significant apoptosis compared to non-silenced cells (P < 0.05); 107 expression changed genes were screened from a total of 49,395 transcripts. Among the total 107 transcripts, 34 transcripts were up-regulated and 73 transcripts were down-regulated. There were five significant differentially expressed genes, including IGFBP4, HOXC8, LMO4, MDK, and CTGF, which correlated with the MeCP2 gene. The methylation frequency of CpG in IGFBP4 gene could achieve 55 %. In conclusion, the differentially expressed IGFBP4, HOXC8, LMO4, MDK, and CTGF genes may be involved in MeCP2 gene-mediated proliferation and apoptosis in osteosarcoma cells.

Keywords

Methyl-CpG-binding protein 2 Osteosarcoma Proliferation Differentially expressed genes 

Notes

Acknowledgments

This work was supported by a grant from the National Natural Science Foundation of China (81001198 and 81372864).

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Gang Meng
    • 1
  • Yi Li
    • 2
  • YangFan lv
    • 1
  • Huanzi Dai
    • 3
  • Xi Zhang
    • 1
  • Qiao-Nan Guo
    • 1
  1. 1.Department of Pathology, Xinqiao HospitalThird Military Medical UniversityChongqingChina
  2. 2.Department of OncologyKunming General Hospital of Chengdu Military CommandKunmingChina
  3. 3.Department of Nephrology, Daping HospitalThird Military Medical UniversityChongqingChina

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