Department of Cardiology, First Affiliated Hospital, Nanchang University, Nanchang, Jiangxi 330033, P.R. China.
Mol Med Rep. 2022 Jul;26(1). doi: 10.3892/mmr.2022.12762. Epub 2022 Jun 3.
The long noncoding RNA LINC00961 plays a crucial role in cancer and cardiovascular diseases. In the present study, the role and underlying mechanism of LINC00961 in endothelial‑mesenchymal transition (EndMT) induced by transforming growth factor beta (TGF‑β), was investigated. Human cardiac microvascular endothelial cells were transfected with LV‑LINC00961 or short hairpin LINC00961 plasmids to overexpress or knock down LINC00961 in the cells, respectively. The cells were then exposed to TGF‑β in serum‑free medium for 48 h to induce EndMT. Flow cytometric analysis, Cell Counting Kit‑8 assay and immunofluorescence staining were performed to examine the cell apoptosis rate, assess cell viability, and identify CD31/α‑SMA double‑positive cells, respectively. Western blotting and reverse transcription‑ quantitative polymerase chain reaction were used to evaluate protein and mRNA expression, respectively. Injury to endothelial cells and EndMT was induced by TGF‑β in a time‑dependent manner. LINC00961 overexpression promoted injury and EndMT, whereas LINC00961 knockdown had the opposite effects. Knockdown of LINC00961 attenuated EndMT and injury to endothelial cells induced by TGF‑β via the PTEN‑PI3K‑AKT pathway. Inhibition of LINC00961 expression may prevent the occurrence of EndMT‑related cardiovascular diseases, such as myocardial fibrosis and heart failure. Therefore, LINC00961 shows potential as a therapeutic target for cardiovascular diseases.
长链非编码 RNA LINC00961 在癌症和心血管疾病中发挥着关键作用。在本研究中，研究了 LINC00961 在转化生长因子β（TGF-β）诱导的内皮-间质转化（EndMT）中的作用及其潜在机制。用 LV-LINC00961 或短发夹 LINC00961 质粒转染人心肌微血管内皮细胞，分别在细胞中转染 LINC00961 过表达或敲低。然后将细胞在无血清培养基中暴露于 TGF-β 中 48 h 以诱导 EndMT。通过流式细胞术分析、细胞计数试剂盒-8 测定和免疫荧光染色分别检测细胞凋亡率、评估细胞活力和鉴定 CD31/α-SMA 双阳性细胞。Western blot 和逆转录-定量聚合酶链反应分别用于评估蛋白质和 mRNA 表达。TGF-β以时间依赖性方式诱导内皮细胞损伤和 EndMT。LINC00961 过表达促进损伤和 EndMT，而 LINC00961 敲低则产生相反的效果。通过 PTEN-PI3K-AKT 通路，敲低 LINC00961 减弱了 TGF-β诱导的 EndMT 和内皮细胞损伤。抑制 LINC00961 的表达可能防止与 EndMT 相关的心血管疾病的发生，如心肌纤维化和心力衰竭。因此，LINC00961 可能成为心血管疾病的治疗靶点。
