CARM1 is required to maintain stemness of amniotic fluid-derived stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies have shown that methylation modification using CARM1-catalyzed histone H3R17/R26 can maintain the stemness of embryonic stem cel s. However, mechanism underlying CARM1 effect on the stemness of amniotic fluid-derived stem cel s is stil unclear. OBJECTIVE:To investigate the function and underlying molecular mechanism of CARM1 to maintain stemness in the amniotic fluid-derived stem cel s. METHODS:Amniotic fluid-derived stem cel s from term pregnancy were isolated and cultured. RT-PCR was used to identify the stem cel mark and CARM1 gene expression. CARM1 expression in amniotic fluid-derived stem cel s was knocked down by using two shRNA. RT-qPCR was used to detect the silencing efficiency, and western blot employed to examine the methylation level of Arginines 17 at N terminus of histone 3 (H3mR17). Moreover, the expression of embryonic stem cel markers, including OCT4, SOX2 and NANOG, were detected. RESULTS AND CONCLUSION:Amniotic fluid-derived stem cel s from term pregnancy could express CARM1 and stem cel markers, including OCT4, SOX2, Nanog and KLF4. Both of the shRNAs could knock down the expression of CARM1 efficiently. When CARM1 was knocked down, the H3mR17 level was decreased and OCT4, SOX2 expression was also reduced, but NANOG expression had no change. Al these indicate that CARM1 is required for amniotic fluid-derived stem cel s to maintain stemness through regulating OCT4 and SOX2 expression.