LncRNA LOC100360491 overexpression inhibits hypoxia-reoxygenation-induced apoptosis of H9c2 cells by upregulating PAK3 transcription

Abstract

Objective To investigate the effect of long noncoding RNA (LncRNA) LOC100360491 on apoptosis of H9c2 cardiomyocytes induced by hypoxia-reoxygenation injury and explore the possible mechanism. Methods H9c2 cells were infected with lentiviral vectors carrying LOC100360491 gene or its inhibitor (shLncRNA) or with a negative control vector, and the changes in LOC100360491 expression in the cells were detected by qRT-PCR. The infected H9c2 cells were exposed to hypoxia for 24 h followed by reoxygenation for 6 h, and early apoptosis and reactive oxygen species (ROS) production of the cells were analyzed by flow cytometry. The expression of pro-caspase-3, cleaved caspase-3, Bax, and Bcl-2 proteins in the cells were detected using Western blotting; the cellular expression levels of PAK3 and CNNM2 mRNA were detected using qRT-PCR. Results The results of qRT-PCR confirmed successful overexpression or inhibition of LOC100360491 in H9c2 cells (P < 0.05). Compared with the cells infected with the control lentiviral vector, the cells with lentivirus-mediated LOC100360491 overexpression showed significantly decreased early apoptosis and ROS production with obviously down-regulated expressions of cleaved caspase-3 and Bax and up-regulated Bcl-2 expression following hypoxia-reoxygenation (P < 0.05). Inhibition of LOC100360491 in H9c2 cells resulted in the opposite changes, which were not significant compared with negative control group. Compared with the cells infected with the negative control vector, the cells with LOC100360491 suppression showed significantly down-regulated expressions of PAK3 and CNNM2 mRNA (P < 0.05), while LOC100360491 overexpression resulted in obviously up-regulated PAK3 expression (P < 0.05) without significantly affecting CNNM2 expression. Conclusion Overexpression of LOC100360491 inhibits apoptosis of H9c2 cells induced by hypoxia-reoxygenation possibly by inhibiting mitochondrial release of ROS through upregulating the transcription of PAK3.

Keywords

• long noncoding rna
• lancrna loc100360491
• myocardial cells
• hypoxia reoxygenation
• apoptosis