DNA demethylation with 5-aza-2'-deoxycytidine induces the senescence-associated secretory phenotype in the immortal fish cell line, EPC

Gene
Kunihiko FutamiTakayuki Katagiri

Abstract

Fish cell lines are known to be immortal and do not show the signs of cellular senescence despite the absence of transformation. Furthermore, high telomerase activities responsible for maintenance of telomere length are detected in many organs in live fish, irrespective of fish age. On the other hand, although it is reported that cytosine methylation at CpG island shores decreases as zebrafish age, the relationship between DNA methylation and cellular senescence in fish has not been explored. In this study, we investigated the induction of cellular senescence and senescence-associated secretory phenotype (SASP) in a fathead minnow Pimephales promelas immortal cell line, Epithelioma papulosum cyprini (EPC) treated with the DNA demethylating agent 5-Aza-2'-deoxycytidine (5-Aza-dC). DNA demethylation by 10 μM of 5-Aza-dC caused cell growth arrest, morphological senescence-like phenotypes and induction of senescence-associated β-galactosidase (SA-β-gal) activity, likely due to a mitotic catastrophe caused by disruption of chromosome segregation. Furthermore, RT-qPCR analyses revealed significant up-regulation of senescence markers such as p53-p21 and p16-Rb pathways as well as several SASP factors in 5-Aza-dC treated cells. Meanwhi...Continue Reading

Citations

Nov 26, 2019·Bioscience, Biotechnology, and Biochemistry·Akifumi ShiomiMasato Umeda

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