Genome-wide analysis of histone H3 lysine 4 trimethylation by ChIP-chip in rat lung fibroblast transdifferentiation
10.3867/j.issn.1000-3002.2016.07.004
- VernacularTitle:采用ChIP-chip技术分析大鼠肺成纤维细胞转分化前后全基因组蛋白H3K4三甲基化水平的改变
- Author:
Suna LIU
;
Wu YAO
;
Lei BAO
;
Juan LI
;
Hongyi ZHANG
;
Jianyong HOU
;
Di WANG
;
Huiting CHEN
;
Changfu HAO
- Publication Type:Journal Article
- Keywords:
fibroblast;
silicon dioxide;
alveolar macrophages;
chromatin co-immunoprecipitation;
histone H3 lysine 4;
methylation
- From:
Chinese Journal of Pharmacology and Toxicology
2016;30(7):728-735
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To analyze trimethylation of genome-wide histone H3 lysine 4(H3K4met3) induced by silicon dioxide(SiO2)through chromatin immunoprecipitation linked to microarrays(ChIP-chip)in lung fibroblast(LF)of rats. METHODS A primary co-culture model of rat alveolar macrophages (AM)and LF in vitro. AM were exposed to 100 mg · L-1 free SiO2 for 24 h,before LF were collected and the phenotype of LF was determined after transdifferentiation by immunohistochemistry. ChIP-chip was used to profile the variations of trimethylation in H3K4 of lung fibroblasts in CpG island regions. ChIP-qPCR was used to validate the microarray results. The mRNA expression of nfib and kpna3 was analyzed by qRT-PCR. RESULTS Totally 1815 (518 increased and 1297 decreased) genes of H3K4met3 displayed significant differences in SiO2 100 mg·L-1 group compared with control group(Cy3/Cy5 value>2.0 or <0.5,NimbleScan V2.5 software). The results of ChIP-qPCR were quite consistent with those of microarray. CONCLUSION There are significant differences in methylation of genome-wide H3K4 between SiO2 100 mg·L-1 group and control group. These novel candidate genes may become potential biomarkers or new interfered targets.
