Zhao L-Y, Li X, Gao L, Xu Y
Medical Treatment Center, Weifang People's Hospital Brain Hospital, Weifang, China.
Eur Rev Med Pharmacol Sci. 2019 Aug;23(3 Suppl):334-340. doi: 10.26355/eurrev_201908_18665.
To explore the effects of long non-coding ribonucleic acid (lncRNA) maternally expressed gene 3 (MEG3) on the proliferation and apoptosis of hypoxic myocardial cells by regulating the expression of forkhead box O1 (FoxO1).
The myocardial A10 cell lines were divided into myocardial cell group (group A), hypoxic myocardial cell group (group B), and hypoxic myocardial cell + transfection with lncRNA MEG3 mimic group (group C). The correlations of the adenosine triphosphate (ATP) concentration, the degree of apoptosis, and the proliferation with FoxO1 and FoxO3a proteins in the cells were observed via ATP assay, Cell Counting Kit-8 (CCK-8) assay, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, Western blotting, and quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR), respectively.
The ATP concentration in myocardial cells was the highest in group A (p<0.05), and it was higher in group B than that in group C (p<0.05). The results of the CCK-8 assay showed that the proliferation rate of the myocardial cells was the highest in group A and the lowest in group C (p<0.05), and it was significantly increased in group B compared with that in group C (p<0.05). The results of the TUNEL assay revealed that the normal cells displayed the purple color, and apoptotic cells displayed the green color. The myocardial cells were arranged orderly, and the number of apoptotic cells was smaller in group A, the number of apoptotic cells was significantly larger in group B than that in group A, and it was the largest in group C (p<0.05). Moreover, the results of Western blotting manifested that the concentrations of FoxO1 and FoxO3 proteins in myocardial cells were the lowest in group A (p<0.05), and they were significantly higher in group C than those in group B (p<0.05). According to the results of qRT-PCR, the mRNA expressions of FoxO1 and FoxO3 in myocardial cells were the lowest in group A (p<0.05), and they were remarkably lower in group B than those in group C (p<0.05).
LncRNA MEG3 can increase the activity of FoxO1 to promote myocardial apoptosis in a hypoxic environment.
通过调节叉头框O1（FoxO1）的表达，探讨长链非编码核糖核酸（lncRNA）母源表达基因3（MEG3）对缺氧心肌细胞增殖和凋亡的影响。
将心肌A10细胞系分为心肌细胞组（A组）、缺氧心肌细胞组（B组）和缺氧心肌细胞+转染lncRNA MEG3模拟物组（C组）。分别通过ATP检测、细胞计数试剂盒-8（CCK-8）检测、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）检测、蛋白质印迹法和定量逆转录-聚合酶链反应（qRT-PCR）观察细胞中三磷酸腺苷（ATP）浓度、凋亡程度以及增殖与FoxO1和FoxO3a蛋白的相关性。
A组心肌细胞中的ATP浓度最高（p<0.05），B组高于C组（p<0.05）。CCK-8检测结果显示，心肌细胞增殖率A组最高，C组最低（p<0.05），B组较C组显著升高（p<0.05）。TUNEL检测结果显示，正常细胞呈紫色，凋亡细胞呈绿色。心肌细胞排列有序，A组凋亡细胞数量较少，B组凋亡细胞数量显著多于A组，C组最多（p<0.05）。此外，蛋白质印迹法结果表明，心肌细胞中FoxO1和FoxO3蛋白浓度A组最低（p<0.05），C组显著高于B组（p<0.05）。根据qRT-PCR结果，心肌细胞中FoxO1和FoxO3的mRNA表达A组最低（p<0.05），B组显著低于C组（p<0.05）。
lncRNA MEG3可增加FoxO1的活性，在缺氧环境中促进心肌细胞凋亡。
