Department of Cardiac Surgery, Zhongshan Hospital of Fudan University, Shanghai, China.
Int J Exp Pathol. 2020 Dec;101(6):196-202. doi: 10.1111/iep.12377. Epub 2020 Oct 14.
Cardiomyocyte differentiation is a multi-step process which involves a number of signalling pathways. microRNAs exhibit regulatory functions in various diseases and are involved in the signalling pathways in multiple physiological processes, but the specific functions of particular mRNAs is often not fully understood. of an example of this is that the role of miR-590-3p in the differentiation of cardiomyocytes remains unclear. In the current study, RT-qPCR was used to determine the expression of miR-590-3p in cardiomyocytes differentiated from the embryonic carcinoma cell line P19CL6. MTT, EdU, caspase-3 activity and flow cytometry assays were performed to examine the influence of miR-590-3p on cell behaviour. A luciferase assay was used to confirm binding between miR-590-3p and PTPN1. Western blotting was used to determine the relationship between the JNK/STAT/NF-kB pathway and PTPN1. The results inferred that miR-590-3p became heavily expressed in differentiated P19CL6. Knockdown miR-590-3p suppressed the cell proliferation while at the same time, accelerated apoptosis. Moreover, PTPN1 was identified as the target of miR-590-3p. More importantly, PTPN1 overexpression activated the JNK/STAT/NF-kB pathway and limited the differentiation of P19CL6. Thus the conclusions from this study are that miR-590-3p has the potential to regulate the proliferation, apoptosis and differentiation of cardiomyocyte P19CL6 in vitro by targeting PTPN1 via the JNK/STAT/NF-kB pathway.
心肌细胞分化是一个多步骤的过程，涉及许多信号通路。microRNAs 在各种疾病中表现出调节功能，并且参与多个生理过程的信号通路，但特定 mRNA 的具体功能通常并不完全清楚。例如，miR-590-3p 在心肌细胞分化中的作用尚不清楚。在本研究中，通过 RT-qPCR 确定了胚胎癌细胞系 P19CL6 分化的心肌细胞中 miR-590-3p 的表达。通过 MTT、EdU、caspase-3 活性和流式细胞术检测来检查 miR-590-3p 对细胞行为的影响。通过荧光素酶测定来确认 miR-590-3p 与 PTPN1 之间的结合。通过 Western blot 来确定 JNK/STAT/NF-kB 通路与 PTPN1 之间的关系。结果推断 miR-590-3p 在分化的 P19CL6 中大量表达。下调 miR-590-3p 抑制细胞增殖，同时加速细胞凋亡。此外，PTPN1 被鉴定为 miR-590-3p 的靶标。更重要的是，PTPN1 的过表达激活了 JNK/STAT/NF-kB 通路，并限制了 P19CL6 的分化。因此，本研究的结论是，miR-590-3p 通过靶向 JNK/STAT/NF-kB 通路靶向 PTPN1，具有调节体外 P19CL6 心肌细胞增殖、凋亡和分化的潜力。
