Mobile website
Basic Research
Hypermethylation of the transmembrane protein125 in lung adenocarcinoma mediates the activation of NF- κB signaling pathway and reduces the sensitivity to decitabine
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To explore the expression of transmembrane protein 125(TMEM125) in lung adenocarcinoma (LUAD) tissues and A549 cells, and the molecular mechanism that affects the proliferation and invasion of A549 cells. Methods: Collected and downloaded the lung adenocarcinoma clinical information and gene expression profile data from The Cancer Genome Atlas (TCGA) database, and analyzed the correlation between the expression of TMEM125 in lung adenocarcinoma and the overall survival of patients.Constructed TMEM125 overexpressing A549 cell line, detected the effect of TMEM125 overexpression on the proliferation and migration of A549 cells by CCK-8 assay and Wound healing assay, and detected the effect of TMEM125 overexpression on the cell cycle and apoptosis of A549 cells by using flow cytometry. WB was used to detect the effect of TMEM125 overexpression on downstream NF- κB signaling pathways and apoptotic proteins. Co-immunoprecipitation (Co-IP) was used to detect the interaction between TMEM125 and NF-κB inhibitor interacting Ras-like 2 (NKIRAS2). TMEM125 overexpressing cells was treated with TNFα（10 ng/ml）and then CCK-8 assay, flow cytometry and WB were used to detect its effects on cell proliferation, apoptosis and NF- κB signaling pathway proteins. A549 cells were treated with demethylation reagent decitabine, and the expression of TMEM125 gene and protein was detected by qPCR and WB. Results: The expression level of TMEM125 mRNA in LUAD tissue was significantly lower than that in normal tissues (P<0.001), the promoter methylation level was significantly higher than that in normal tissues (P<0.001), and the overall survival of patients with low and medium expression was significantly lower than that of patients with high expression (P<0.001).Overexpression of TMEM125 inhibited the proliferation and migration of A549 cells (P<0.01), increased cell G2/M phase and promoted cell apoptosis (P<0.01). Overexpression of TMEM125 could interact with NKIRAS2 and inhibit the activity of NF- ΚB (P<0.01).Treatment of A549 cells with decitabine could promote the expression of TMEM125 and inhibit cell proliferation (P<0.01).Conclusion:Promoter hypermethylation inhibits TMEM125 gene expression, leading to a decline in its function of inhibiting NF-κB activity and inhibiting cell proliferation and therefore reduce the sensitivity to decitabine, and resulting in the decreased sensitivity to decitabine.
Keywords:
Project Supported:
Project supported by the Health Industry Scientific Research Project of Hainan Province（No.19A200014）
Clc Number:
