Effect of LINC00612 on Hypoxia/Reoxygenation Induced Cardiomyocytes Injury by Targeting miR-30d
CHENG Lin1, FENG Li2, ZHANG Hua3, XU Aiguo1*
In order to investigate the effect of LINC00612 (long intergenic non-coding RNA 00612) targeting miR (microRNA)-30d on hypoxia/reoxygenation injury of cardiomyocytes, the relative levels of LINC00612 and miR-30d in the plasma of patients with myocardial infarction were detected by real-time quantitative PCR. Meanwhile, a rat cardiomyocyte H9C2 hypoxia/reoxygenation injury model was established. The pcDNA (empty vector plasmid), pcDNA-LINC00612 (LINC00612 overexpression vector), anti-miR-NC (miRNA inhibitor negative control), anti-miR-30d (miR-30d inhibitor), pcDNA-LINC00612+miR-30d minics were transfected into H9C2 cells, respectively. After hypoxia/reoxygenation treatment, CCK-8 was used to detect cell viability. Flow cytometry was applied to detect cell apoptosis, and commercial kits were employed to detect activity of SOD (superoxide dismutation) in cells and the levels of CK (creatine kinase), LDH (lactate dehydrogenase) in cell culture fluid. In this study, compared with healthy controls, the relative level of LINC00612 in the plasma of patients with myocardial infarction was significantly reduced (P<0.05), and the relative level of miR-30d was significantly increased (P<0.05). Hypoxia/reoxygenation treatment significantly down-regulated LINC00612 expression (P<0.05), upregulated miR-30d expression (P<0.05), decreased cell viability and SOD activity (P<0.05) and increased apoptosis rate of H9C2 cells, as well as increased the levels of CK and LDH in cell culture medium (P<0.05). LINC00612 overexpression or miR-30d inhibition significantly increased cell viability and SOD activity (P<0.05), and reduced the apoptosis rate and the levels of CK and LDH in cell culture (P<0.05). miR-30d overexpression significantly reduced cell viability and SOD activity (P<0.05), and increased apoptosis rate, as well as increased and the levels of CK and LDH in cell culture medium (P<0.05). Overexpression of miR-30d could significantly reduce the effects of LINC00612 overexpression on the viability, apoptosis and oxidative injury of hypoxia/reoxygenation cardiomyocytes (P<0.05). In conclusion, LINC00612 can reduce hypoxia/reoxygenation induced cardiomyocyte apoptosis and oxidative stress injury by targeting miR-30d.
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