期刊
ONCOTARGET
卷 7, 期 44, 页码 71922-71936出版社
IMPACT JOURNALS LLC
DOI: 10.18632/oncotarget.12431
关键词
microRNA-130a; PTEN; PI3K/Akt/eNOS signaling pathway; coronary artery endothelial cells; cell injury
资金
- National Natural Science Foundation of China Youth Science Fund [51103059]
- Natural Science Foundation of Jilin Province [201115071, 20140101054JC]
- Leading Talents and Team Project of Science [20150519025JH]
- Guiding Fund for Strategic Adjustment of Economic Structure Special Projects of Jilin Province [2015Y030-3]
- Science and Technology Research Projects of the 12th Five-Year Plan for Jilin Province Department of Education [[2014] B016, [2015] 524]
- Industrial technology research and development project of Jilin provincial industry innovation special fund project (part of high tech industry) [2016C044-2]
Our study aims to investigate the roles of microRNA-130a (miR-130a) in human coronary artery endothelial cells (HCAECs) injury and inflammatory responses by targeting PTEN through the PI3K/Akt/eNOS signaling pathway. HCAECs were treated with 1.0 mmol/L homocysteine (HCY) and assigned into eight groups: the blank group, the negative control (NC) group, the miR-130a mimics group, the miR-130a inhibitors group, the si-PTEN group, the Wortmannin group, the miR-130a inhibitors + si-PTEN group and the miR-130a mimics + Wortmannin group. Luciferase reporter gene assay was used to validate the relationship between miR-130a and PTEN. The expressions of miR-130a, PTEN and PI3K/Akt/eNOS signaling pathway-related proteins were detected by qRT-PCR assay and Western blotting. MTT assay and Hoechst 33258 staining were adopted to testify cell growth and apoptosis. The NO kit assay was used to detect the NO release. ELISA was conducted to measure serum cytokine levels. Luciferase reporter gene assay confirmed the target relationship between miR-130a and PTEN. Compared with the blank and NC groups, the miR-130a mimics and si-PTEN groups showed significant increases in the expressions of PI3K/Akt/eNOS signaling pathway-related proteins, cell viability and the NO release, while serum cytokine levels and cell apoptosis were decreased; by contrast, an opposite trend was observed in miR-130a inhibitors and Wortmannin groups. However, no significant difference was found in the miR-130a inhibitors + si-PTEN and miR-130a mimics + Wortmannin groups when compared with the blank group. These results indicate that miR-130a could alleviate HCAECs injury and inflammatory responses by down-regulating PTEN and activating PI3K/Akt/eNOS signaling pathway.
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