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Preliminary screening of differentially expressed genes involved in methyl-CpG-binding protein 2 gene-mediated
1Department of Pathology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China.
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.
Insights
Methyl-CpG-binding protein 2 (MeCP2) silencing inhibits osteosarcoma cell proliferation and induces apoptosis. Five key genes, including IGFBP4, were identified as potential therapeutic targets for osteosarcoma treatment.
Area of Science:
- Molecular Biology
- Oncology
- Epigenetics
Background:
- Methyl-CpG-binding protein 2 (MeCP2) plays a crucial role in brain development and is implicated in various cancers and neurodevelopmental disorders.
- Understanding MeCP2's role in osteosarcoma is vital for identifying novel therapeutic strategies.
Purpose of the Study:
- To screen MeCP2-related differentially expressed genes in osteosarcoma.
- To identify potential therapeutic targets for osteosarcoma based on MeCP2 gene expression.
Main Methods:
- Cell proliferation was assessed using the CCK8 assay in SaOS2 and U2OS cells.
- Apoptosis was analyzed via flow cytometry (Annexin V-APC/7-ADD).
- Gene expression profiling identified differentially expressed genes using gene chip technology.
Main Results:
- MeCP2 silencing significantly reduced osteosarcoma cell proliferation and induced apoptosis (P < 0.05).
- Gene chip analysis identified 107 differentially expressed genes (34 upregulated, 73 downregulated) from 49,395 transcripts.
- Five significant differentially expressed genes (IGFBP4, HOXC8, LMO4, MDK, CTGF) correlated with MeCP2, with IGFBP4 showing 55% CpG methylation frequency.
Conclusions:
- MeCP2 influences osteosarcoma cell proliferation and apoptosis.
- The identified genes (IGFBP4, HOXC8, LMO4, MDK, CTGF) are potential mediators of MeCP2's effects in osteosarcoma.
- These genes represent promising therapeutic targets for osteosarcoma treatment.
