Background/Aims: Myocardial ischemia (MI) is a serious threat to human health. Circular RNAs (circRNAs) play an important role in many diseases including MI. The effect and mechanism of circDENND2A in MI have not been studied. Methods: We used oxygen glucose deprivation (OGD) treatment to simulate MI in vitro. We detected circDENND2A and microRNA (miR)-34a levels by RT-qPCR. The transfection process used INTERFER and jetPRIME. Cell growth indexes including viability, apoptosis, and migration were detected by CCK8, flow cytometry, and transwell assays, respectively. In addition, the Bax, Cleaved-Caspase-3, matrix metalloproteinase (MMP)-2, MMP-9 and pathway-related protein levels were tested by Western blot. Results: OGD upregulated circDENND2A expression in H9c2 cells. Overexpression of circDENND2A enhanced cell viability and migration but declined apoptosis under OGD. Silenced circDENND 2A played the opposite effects. circDENND2A negatively regulated miR-34a. miR-34a overexpression weakened the protective effects of circDENND2A in OGD-injury. Moreover, we considered circDENND2A and miR-34a may work via β-catenin and Ras/Raf/MEK/ERK pathways. Conclusion: circDENND2A overexpression enhanced OGD-inhibited cell viability and migration but declined OGD-promoted apoptosis by downregulating miR-34a and via β-catenin and Ras/Raf/MEK/ERK pathways.
Background/Aims: Myocardial ischemia (MI) is a serious threat to human health. Circular RNAs (circRNAs) play an important role in many diseases including MI. The effect and mechanism of circDENND2A in MI have not been studied. Methods: We used oxygen glucose deprivation (OGD) treatment to simulate MI in vitro. We detected circDENND2A and microRNA (miR)-34a levels by RT-qPCR. The transfection process used INTERFER and jetPRIME. Cell growth indexes including viability, apoptosis, and migration were detected by CCK8, flow cytometry, and transwell assays, respectively. In addition, the Bax, Cleaved-Caspase-3, matrix metalloproteinase (MMP)-2, MMP-9 and pathway-related protein levels were tested by Western blot. Results: OGD upregulated circDENND2A expression in H9c2 cells. Overexpression of circDENND2A enhanced cell viability and migration but declined apoptosis under OGD. Silenced circDENND 2A played the opposite effects. circDENND2A negatively regulated miR-34a. miR-34a overexpression weakened the protective effects of circDENND2A in OGD-injury. Moreover, we considered circDENND2A and miR-34a may work via β-catenin and Ras/Raf/MEK/ERK pathways. Conclusion: circDENND2A overexpression enhanced OGD-inhibited cell viability and migration but declined OGD-promoted apoptosis by downregulating miR-34a and via β-catenin and Ras/Raf/MEK/ERK pathways.
Authors: Mehran Haidari; Wei Zhang; Amy Caivano; Zhenping Chen; Leila Ganjehei; Ahmadreza Mortazavi; Christopher Stroud; Darren G Woodside; James T Willerson; Richard A F Dixon Journal: J Biol Chem Date: 2012-07-25 Impact factor: 5.157