过表达miR-32削弱多氯联苯抑制P19细胞向心肌细胞的分化  

Overexpression of micro RNA-32 weakens the inhibition of polychlorinated biphenyls to the differentiation of P19 cells into cardiomyocytes

作　　者：张松雨[1] 李燕[1] 刘江波[1] 李纲[1] 

机构地区：[1]郑州大学附属南阳医院,南阳市中心医院心内科,河南省南阳市473009

出　　处：《中国组织工程研究》2016年第23期3464-3469,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:多氯联苯对P19细胞分化成心肌细胞有抑制作用。miR NAs在多氯联苯抑制P19细胞分化成心肌细胞过程中发挥了重要的作用。目的:探索mi R-32在调节多氯联苯对P19细胞分化成心肌细胞中的作用。方法:将P19细胞与1%二甲基亚砜和2.5μmol/L多氯联苯共孵育,通过Western blot鉴定分化标志物α-actinin、desmin、cT nI以及早期心脏发育的重要转录因子GATA4的变化。qR T-PCR鉴定多氯联苯对miR-32表达的影响。应用基因重组技术成功构建了小鼠miR-32真核表达载体,将其转染至P19细胞,与2.5μmol/L多氯联苯和1%二甲基亚砜共孵育,通过Western blot实验观察分化相关蛋白表达。结果与结论:(1)多氯联苯降低了miR-32的表达水平,抑制P19细胞向心肌细胞的分化;(2)成功构建了小鼠miR-32真核表达载体,建立了稳定过表达miR-32的P19细胞系;(3)过表达mi R-32发挥了削弱多氯联苯对P19细胞向心肌细胞分化的抑制作用。BACKGROUND： Polychlorinated biphenyls inhibit the differentiation of P19 cells into cardiomyocytes. In the meanwhile, micro RNAs play an important role in regulating cell differentiation. OBJECTIVE： To explore the effect of micro RNA-32（miR-32） on the polychlorinated biphenyls-inhibited differentiation of P19 cells into cardiomyocytes. METHODS： P19 cells were co-cultured with 2.5 μmol/L polychlorinated biphenyls and 1% dimethyl sulfoxide. Afterwards, α-actinin, desmin, c TnI and GATA4 were identified by western blot assay. Wddwxrof polychlorinated biphenyls on the expression of mi R-32 was evaluated by real-time PCR. Mouse eukaryotic vector expressing mi R-32 was constructed by gene recombination technology, and transfected into P19 cells followed by co-cultured with 2.5 μmol/L polychlorinated biphenyls and 1% dimethyl sulfoxide. Then, expressions of differentiation-related proteins were detected by western blot assay. RESULTS AND CONCLUSION： Polychlorinated biphenyls inhibited the differentiation of P19 cells into cardio myocytes and decreased the miR-32 expression. Cell lines overexpressing mi R-32 was successfully established in mice. Furthermore, we found that overexpression of mi R-32 weakens the inhibition of polychlorinated biphenyls to the differentiation of P19 cells into cardiomyocytes.

关 键 词：微RNAS 多氯联苯化合物 肌细胞 心脏 细胞分化 组织工程 干细胞 分化 多氯联苯 miR-32 P19细胞 心肌细胞 过表达 

分 类 号：R394.2[医药卫生—医学遗传学]