摘要
目的:探究环状RNA NCX1(circNCX1)对阿霉素(又称多柔比星,DOX)诱导的心肌细胞凋亡的调节作用。方法:通过RT-qPCR检测经DOX处理的H9c2细胞和注射DOX的小鼠心肌组织中circNCX1表达量的变化;TUNEL染色和凋亡相关蛋白caspase-3/-7的活性检测试剂盒检测细胞凋亡水平;台盼蓝染色检测细胞存活率;RNA pull-down实验验证circNCX1与其下游微小RNA-103-3p(miR-103-3p)的直接结合。结果:DOX诱导的H9c2细胞及小鼠心肌组织中circNCX1升高,并且DOX可诱导心肌细胞凋亡,表现为心肌细胞TUNEL阳性率及caspase-3/-7活性升高。抑制circNCX1表达能够显著减少DOX诱导的心肌细胞TUNEL阳性率,并且降低caspase-3/-7的活性。circNCX1通过直接结合miR-103-3p抑制其表达。此外,过表达miR-103-3p同样能够抑制DOX诱导的心肌细胞死亡和caspase-3/-7活性。结论:circNCX1通过靶向miR-103-3p促进DOX诱导的心肌细胞凋亡。
AIM:To explore the role of circular RNA transcribed from the sodium/calcium exchanger 1 gene(circNCX1)in doxorubicin(DOX)-induced cardiac myocyte apoptosis.METHODS:The circNCX1 expression levels in DOX-treated H9c2 cells and mouse hearts were detected by RT-qPCR.TUNEL assay and caspase-3/-7 activity determination were performed to assess apoptosis,while trypan blue staining was applied to measure cell viability.In addition,RNA pull-down assay was performed to verify the interaction of circNCX1 and miR-103-3p.RESULTS:circNCX1 in cardiac myocytes was up-regulated under DOX treatment in vitro and in vivo.DOX-induced H9c2 cell apoptosis was characterized by increases in TUNEL positive rate and caspase-3/-7 activity.circNCX1 inhibition significantly attenuated DOXinduced H9c2 cells apoptosis,and suppressed caspase-3/-7 activity.circNCX1 directly bound to miR-103-3p and suppressed its expression.Additionally,miR-103-3p mimic inhibited DOX-induced H9c2 cell apoptosis and caspase-3/-7 activity.CONCLUSION:circNCX1 promotes DOX-induced H9c2 cell apoptosis via targeting to miR-103-3p.
作者
法鸿鸽
李萌阳
常文光
肖丹丹
王建勋
FA Hong-ge;LI Meng-yang;CHANG Wen-guang;XIAO Dan-dan;WANG Jian-xun(School of Basic Medical Sciences,Qingdao University,Qingdao University,Qingdao 266021,China;Institute for Translational Medicine,Qingdao University,Qingdao 266021,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第3期450-457,共8页
Chinese Journal of Pathophysiology
基金
山东省自然科学杰出青年基金资助项目(No.JQ201815)
青岛市民生科技计划项目(No.18-6-1-80-nsh)。
