Expression And Methylation Of PRDM2、PRDM5、PRDM16Gene In Lung Cancer Cell Lines

Objective In order to study of expression and methylation status of PRDM2、PRDM5、RDM16gene in A549(lung adenocarcinoma cancer cell lines), HTB-182(lungsquamous cell carcinoma cell lines).Methods Cultured A549(lung adenocarcinoma cell lines), HTB-182(lung squamouscell carcinoma cell lines) in vitro,with HBE (normal bronchial cell lines) as control celllines, we detect methylation state of PRDM2、PRDM5、PRDM16gene by MSP, detectmRNA expression level of PRDM2、PRDM5、PRDM16gene by RT-PCR, detect proteinexpression level of PRDM2、PRDM5、PRDM16gene by Western-blot test；with differentconcentration gradient’smethylation drugs (5-aza-2dc) we demethylated the lung cancercell lines, observed cell growth curve and detect methylation status changes mRNA、protein expression level changes of PRDM2、PRDM5, PRDM16gene before and afterprocessingResults1. There is high methylation in A549(lung adenocarcinoma cancer cell lines) PRDM2、PRDM5、PRDM16gene；there is high methylation in HTB-182(lung squamous cellcarcinoma cell line) PRDM2、PRDM5、PRDM16gene；there is low methylation inHTB-182(lung squamous cell carcinoma cell line) PRDM2、PRDM5、PRDM16gene.2. After different concentrations demethylation drugs (5-aza-2dc) processing,PRDM2、PRDM5、PRDM16gene methylation degree gradually weakened in A549(lungadenocarcinoma cell line), PRDM2、 PRDM5、 PRDM16gene methylation degreegradually weakened in turn in HTB-182(lung squamous carcinoma cell line), PRDM2、PRDM5、PRDM16gene methylation degree is no significant change in HBE (normalbronchial epithelium cell lines)With the demethylating drug concentration increases,PRDM2、PRDM5、PRDM16genemethylation degree gradually weakened, mRNA and protein expression level gradually rise in A549(lung adenocarcinoma cell line), HTB-182(lung squamous cell carcinomacell line)，with the demethylating drug concentration increases,PRDM2、PRDM5、PRDM16gene methylation degree, mRNA and protein expression level did not changesignificantly in HBE (normal bronchial epithelial cell line).3.MTT results showed that through different concentrations’s demethylation drugs(5-aza-2dc) processing，between A549(lung adenocarcinoma cell line), HTB-182(lungsquamous cell carcinoma cell line) proliferation inhibition and demethylationdrugs(5-aza-2dc) concentration was positively correlated,with HBE (normal bronchialepithelium cell lines) proliferation caused by5-aza-2dc did not show significant inhibitoneffect.ConclusionsPRDM2、PRDM5、PRDM16gene exists methylation in A549(lung adenocarcinomacell line), HTB-182(lung squamous cell carcinoma cell line), and gene methylation led toa decline to mRNA and protein expression level of PRDM2、PRDM5、PRDM16gene.Demethylation drugs (5-aza-2dc) be able to reverse PRDM2、PRDM5、PRDM16gene methylation status and can restore PRDM2、PRDM5、PRDM16gene mRNA andprotein expression level.