山西医科大学学报2025，Vol.56Issue(5)：461-468,8.DOI:10.13753/j.issn.1007-6611.2025.05.001
氯乙醛致HepG2细胞染色体损伤及DNA甲基化相关机制研究
Mechanisms related to chromosomal damage and DNA methylation in HepG2 cells induced by chloroacetaldehyde
摘要
Abstract
Objective To explore the mechanisms related to chromosomal damage of HepG2 cells and DNA methylation induced by chloroacetaldehyde.Methods This study conducted in vitro experiments on human hepatocellular carcinoma HepG2 cells from two aspects:exposure to chloroacetaldehyde and intervention with the methyltransferase inhibitor(SGI-1027).The experiment on chloroa-cetaldehyde exposure was divided into blank control group,low chloroacetaldehyde group(2.25 μmol/L),medium chloroacetaldehyde group(4.5 μmol/L),and high chloroacetaldehyde group(9 μmol/L),and the cells were exposed to the agents for 24 h and 48 h,re-spectively.SGI-1027 intervention experiment was divided into solvent control group(DMSO),SGI-1027 group(10 μmol/L),chloroa-cetaldehyde group(4.5 μmol/L),and chloroacetaldehyde+SGI-1027 group(4.5 μmol/L chloroacetaldehyde+10 μmol/L SGI-1027),and the cells were exposed to the agents for 24 h.CCK-8 assay was used to detect the effects of different concentrations of chloroacetal-dehyde and SGI-1027 on the cell viability.Immunofluorescence was used to detect the fluorescence intensity of γ-H2AX to reflect the degree of DNA double-strand breaks.Western blot was used to detect the protein expression levels of DNA damage repair-related genes BRIP1,TOP3,RAD52,and CtIPs.Results After HepG2 cells were exposed to chloroacetaldehyde at various concentrations for 24 h and 48 h,the fluorescence intensity of γ-H2AX increased,and it increased with the increase of chloroacetaldehyde concentration(P＜0.05).The expression level of BRIP1 protein showed no obvious change after 24 h exposure,while it was higher in high-dose group than in control group after 48 h exposure(P＜0.05).After 24 h and 48 h exposure to high chloroacetaldehyde,the expression levels of TOP3 protein showed a downward trend(P＜0.05).The expression levels of RAD52 and CtIP proteins increased after exposure to low chloroacetaldehyde(P＜0.05),while the expression of CtIP protein decreased after 24 h exposure to high chloroacetaldehyde(P＜0.05).Compared with chloroacetaldehyde group,the fluorescence intensity of γ-H2AX decreased in chloroacetaldehyde+SGI-1027 group(P＜0.05),the expressions of CtIP and BRIP1 proteins increased significantly(P＜0.05),while the expressions of RAD52 and TOP3 pro-teins decreased significantly(P＜0.05).Conclusion Chloroacetaldehyde can cause DNA breaks in HepG2 cells,which is related to the low expression of DNA damage repair genes RAD52,TOP3,and CtIP at the protein level.The low expression of CtIP is related to hypermethylation.关键词
氯乙烯/氯乙醛/DNA损伤修复/DNA甲基化/DNA甲基转移酶/SGI-1027Key words
vinyl chloride/chloroacetaldehyde/DNA damage repair/DNA methylation/DNA methyltransferase/SGI-1027分类
医药卫生引用本文复制引用
王静,赵晓田,李树元,王龙美,仇玉兰..氯乙醛致HepG2细胞染色体损伤及DNA甲基化相关机制研究[J].山西医科大学学报,2025,56(5):461-468,8.基金项目
国家自然科学基金资助项目(81773405) （81773405）
