毛壳素对绒山羊ADSCs组蛋白甲基化修饰的影响研究

doi:10.11843/j.issn.0366-6964.2022.07.034

Abstract

Abstract: The study aimed to explore the effects of chaetocin on histone methylation modification in adipose-derived stem cells (gADSCs) of cashmere goat. In this study, gADSCs were treated with different concentrations of chaetocin for different durations, and the concentration and duration of chaetocin treatment that had the least effects on cell viability and the greatest inhibition on histone methylation in gADSCs were screened. The optimal concentration and duration of chaetocin treatment was used as the experimental group, and the chaetocin-free treatment was used as the control group. The expression of H3K9 me2 and H3K9 me3 methylation-related enzymes and embryonic development pluripotency genes were detected by RT-qPCR, and the effects of chaetocin on histone H3K9 me2 and H3K9 me3 proteins expression were observed as well. The results showed that there was little effect on cell viability when gADSCs were treated with 20 nmol·L^-1 chaetin for 48 h, and the expression of histone methyltransferase G9A significantly reduced (P<0.05), the optimal chaetin treatment concentration and duration was confirmed. RT-qPCR results showed that the expression of H3K9 me2 and H3K9 me3 methyltransferase EHMT1, EHMT2, SUV39H1 and SUV39H2 significantly reduced(P<0.05), and the expression of H3K9 me2 and H3K9 me3 demethylase KDM3A, KDM3B, KDM4B and KDM 4Dsignificantly increased(P<0.05),the expression of multipotency-related genes SOX2, OCT4, NANOG increased in experimental group. The results of immunofluorescence and WB experiments showed that the H3K9 me2 protein level did not significantly change, while the H3K9 me3 protein level significantly reduced(P<0.05) in the experimental group. The modification of histone H3K9 me2 and H3K9 me3 in gADSCs treated with 20 nmol·L^-1 chaetocin for 48 h could be reduced significantly, and the expression of multipotency-related genes could be increased.

Key words: chaetocin, adipose-derived stem cells, epigenetic modification, H3K9 me2, H3K9 me3

CLC Number:

S827.2

SUN Hao, ZHE Xiaoshu, ZHANG Wenqi, HAO Fei, LI Wennan, LIU Jie, LIU Dongjun. Effects of Chaetocin on Histone Methylation Modification of Cashmere Goats ADSCs[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2380-2389.

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Effects of Chaetocin on Histone Methylation Modification of Cashmere Goats ADSCs

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