Purpose: Ischemia-reperfusion (I/R) injury exacerbates myocardial cell death (including apoptosis and necrosis), leading to complications such as arrhythmias, myocardial stenosis, microvascular obstruction and heart f...Purpose: Ischemia-reperfusion (I/R) injury exacerbates myocardial cell death (including apoptosis and necrosis), leading to complications such as arrhythmias, myocardial stenosis, microvascular obstruction and heart failure, and it is particularly important to seek new strategies to mitigate reperfusion injury. In this paper, we will investigate whether atorvastatin can alleviate myocardial ischemia-reperfusion injury and verify its molecular mechanism. Methods: We successfully constructed a hypoxia-reperfusion (H/R) H9c2 cell model and transfected miR-26a-5p mimic, miR-26a-5p inhibitor and its negative control NC-mimic or NC-inhibitor into H9c2 cells using a transfection kit. The expression of miR-26a-5p and FOXO1 were detected by RT-qPCR assay, the expression of related proteins by Western blot assay, the cell viability of H9c2 cells by CCK-8 assay, the apoptosis rate of H9c2 cells by flow cytometry, the CK and LDH activity in cells by CK and LDH assay kits. The targeting relationship between miR-26a-5p and FOXO1 was verified by dual luciferase reporter gene assay. Results: MiR-26a-5p expression was decreased in H/R-induced cells and FOXO1 expression was increased in H/R-induced cells. Atorvastatin alleviated H/R injury in cardiomyocytes and was most effective at a concentration of 1 μM. Atorvastatin alleviated H/R injury in cardiomyocytes by upregulating miR-26a-5p expression, miR-26a-5p and FOXO1 were negatively regulated by targeting. Conclusion: Atorvastatin can alleviate H/R injury in cardiomyocytes by regulating miR-26a-5p/FOXO1.展开更多
Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-sp...Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-special 5(GAS5)on proliferation and apoptosis of hepatocellular carcinoma(HCC)cells through miR-26a-5p action.Methods Expression levels of GAS5 were detected in cancerous and paracancerous tissue of 80 HCC patients by RT-qPCR.The starBase tool predicted that GAS5 had binding sites for the miRNA miR-26a-5p,which was also highly expressed in HCC tissue.The relationship between GAS5 and miR-26a-5p was confirmed using a luciferase reporter assay.The role of these lncRNAs was further explored by transfecting plasmids into SMMC-7721 cells and classifying the cells as follows:NC group,GAS5 group,anti-miR-26a-5p group,and GAS5+miR-26a-5p group.Cell proliferation,cell cycle,and apoptosis were detected in each group.The relationship between miR-26a-5p and phosphatase and tensin homolog deleted on chromosome 10(PTEN)was analyzed by TargetScan database prediction and luciferase reporter assay.Western blotting was used to quantify PTEN,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),cyclin D1,and human P27 protein(P27).Results GAS5 was downregulated,while miR-26a-5p was upregulated in HCC tissue compared to in paracancerous tissue.High GAS5 levels and low miR-26a-5p levels inhibited cell proliferation,increased the number of G0/G1 phase cells,promoted cell apoptosis,promoted PTEN and P27 expression,and inhibited PI3K,P-Akt,and cyclin D1 expression at the protein level.Upregulation of miR-26a-5p attenuated the effects of GAS5 upregulation on the proliferation,cell cycle,and apoptosis of HCC cells and on the expression of PTNE/PI3K/Akt signaling pathway-related proteins.Conclusion Low GAS5 levels regulate the proliferation and apoptosis of HCC cells via the PTNE/PI3K/Akt signaling pathway and are linked to upregulation of miR-26a-5p.展开更多
文摘Purpose: Ischemia-reperfusion (I/R) injury exacerbates myocardial cell death (including apoptosis and necrosis), leading to complications such as arrhythmias, myocardial stenosis, microvascular obstruction and heart failure, and it is particularly important to seek new strategies to mitigate reperfusion injury. In this paper, we will investigate whether atorvastatin can alleviate myocardial ischemia-reperfusion injury and verify its molecular mechanism. Methods: We successfully constructed a hypoxia-reperfusion (H/R) H9c2 cell model and transfected miR-26a-5p mimic, miR-26a-5p inhibitor and its negative control NC-mimic or NC-inhibitor into H9c2 cells using a transfection kit. The expression of miR-26a-5p and FOXO1 were detected by RT-qPCR assay, the expression of related proteins by Western blot assay, the cell viability of H9c2 cells by CCK-8 assay, the apoptosis rate of H9c2 cells by flow cytometry, the CK and LDH activity in cells by CK and LDH assay kits. The targeting relationship between miR-26a-5p and FOXO1 was verified by dual luciferase reporter gene assay. Results: MiR-26a-5p expression was decreased in H/R-induced cells and FOXO1 expression was increased in H/R-induced cells. Atorvastatin alleviated H/R injury in cardiomyocytes and was most effective at a concentration of 1 μM. Atorvastatin alleviated H/R injury in cardiomyocytes by upregulating miR-26a-5p expression, miR-26a-5p and FOXO1 were negatively regulated by targeting. Conclusion: Atorvastatin can alleviate H/R injury in cardiomyocytes by regulating miR-26a-5p/FOXO1.
文摘Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-special 5(GAS5)on proliferation and apoptosis of hepatocellular carcinoma(HCC)cells through miR-26a-5p action.Methods Expression levels of GAS5 were detected in cancerous and paracancerous tissue of 80 HCC patients by RT-qPCR.The starBase tool predicted that GAS5 had binding sites for the miRNA miR-26a-5p,which was also highly expressed in HCC tissue.The relationship between GAS5 and miR-26a-5p was confirmed using a luciferase reporter assay.The role of these lncRNAs was further explored by transfecting plasmids into SMMC-7721 cells and classifying the cells as follows:NC group,GAS5 group,anti-miR-26a-5p group,and GAS5+miR-26a-5p group.Cell proliferation,cell cycle,and apoptosis were detected in each group.The relationship between miR-26a-5p and phosphatase and tensin homolog deleted on chromosome 10(PTEN)was analyzed by TargetScan database prediction and luciferase reporter assay.Western blotting was used to quantify PTEN,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),cyclin D1,and human P27 protein(P27).Results GAS5 was downregulated,while miR-26a-5p was upregulated in HCC tissue compared to in paracancerous tissue.High GAS5 levels and low miR-26a-5p levels inhibited cell proliferation,increased the number of G0/G1 phase cells,promoted cell apoptosis,promoted PTEN and P27 expression,and inhibited PI3K,P-Akt,and cyclin D1 expression at the protein level.Upregulation of miR-26a-5p attenuated the effects of GAS5 upregulation on the proliferation,cell cycle,and apoptosis of HCC cells and on the expression of PTNE/PI3K/Akt signaling pathway-related proteins.Conclusion Low GAS5 levels regulate the proliferation and apoptosis of HCC cells via the PTNE/PI3K/Akt signaling pathway and are linked to upregulation of miR-26a-5p.
