F9 EC 세포에서 레티노산에 의해 유도되는 Hoxc 유전자의 발현에 히스톤 메틸화가 미치는 영향

Abstract

Department of Anatomy, Embryology Laboratory, Brain Korea 21 Pl us project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, KoreaReceived December 1, 2014/Revised March 11, 2015/Accepted March 12, 2015Hox genes encode a highly conserved family of homeodomain-containing transcription factors control-ling vertebrate pattern formation along the anteroposterior body axis during embryogenesis. Retinoic acid (RA) is a key morphogen in embryogenesis and a critical regulator of both adult and embryonic cellular activity. Specifically, RA regulates Hox gene expression in mouse- or human-derived embry-onic carcinoma (EC) cells. Histone modification has been reported to play a pivotal role in the process of RA-induced gene expression and cell differentiation. As histone modification is thought to play an essential role in RA-induced Hox gene expression, we examined RA-induced initiation of collinear ex-pression of Hox genes and the corresponding histone modifications in F9 murine embryonic ter-atocarcinoma (EC) cells. Hox expression patterns and histone modifications were analyzed by semi- quantitative RT-PCR, RNA-sequencing, and chromatin immuno-precipitation (ChIP)-PCR analyses. The Hoxc4 gene (D0) was initiated earlier than the Hoxc5 to –c10 genes (D3) upon RA treatment (day 0 [D0], day 1 [D1], and day 3 [D3]). The Hox nonexpressing D0 sample had a strong repressive marker, H3K27me3, than the D1 and D3 samples. In the D1 and D3 samples, reduced enrichment of the H3K27me3 marker was observed in the whole cluster. The active H3K4me3 marker was closely asso-ciated with the collinear expression of Hoxc genes. Thus, the Hoxc4 gene (D1) and all Hoxc genes (D3) expressed H3K4me3 upon transcription activation. In conclusion, these data indicated that removing H3K27me3 and acquiring H3K4me3 regulated RA-induced Hoxc gene collinearity in F9 cells. Keywords :