Dong Li1, Yi An2. 1. Department of Emergency, Linyi People's Hospital, Linyi 276000, Shandong Province, China. 2. Department of Cardiovascular Medicine, Affiliated Hospital of Qingdao University, Qingdao 266071, Shandong Province, China. Electronic address: Any2018@qdu.edu.cn.
Abstract
BACKGROUND: It is reportedly demonstrated that miR-135a-5p plays a critical role in cancer cells, macrophages, and endothelia cells. However, little is known concerning the function of miR-135a-5p in vascular smooth muscle cells (VSMCs) and atherosclerosis (AS). METHODS: Human VSMCs and male C57BL/6 mice were used for establishing AS cell models and animal models. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expressions of miR-135a-5p, forkhead box O1 (FOXO1) mRNA, and Janus kinase 2 (JAK2) mRNA. CCK-8, BrdU, and Transwell assays were used to detect cell migration and proliferation. Cell cycle and apoptosis were analyzed using flow cytometry. The interactions among miR-135a-5p, FOXO1 and JAK2 were validated employing Western blot, qRT-PCR and Luciferase reporter gene assay. RESULTS: The expression of miR-135a-5p was significantly decreased in serum samples of AS patients, VSMCs treated with ox-LDL and AS mice models. The overexpression of miR-135a-5p induced VSMCs cycle arrest and apoptosis, and inhibited proliferation and migration. Further experiments confirmed that miR-135a-5p could target and repress FOXO1/CyclinD1 and JAK2/STAT3 pathway. Additionally, the associations among miR-135a-5p, FOXO1/Cyclin D1 and JAK2/STAT3 were validated using animal models. CONCLUSION: MiR-135a-5p suppresses VSMCs proliferation and migration induced by ox-LDL via targeting and activating FOXO1/Cyclin D1 and JAK2/STAT3 signaling pathways.
BACKGROUND: It is reportedly demonstrated that miR-135a-5p plays a critical role in cancer cells, macrophages, and endothelia cells. However, little is known concerning the function of miR-135a-5p in vascular smooth muscle cells (VSMCs) and atherosclerosis (AS). METHODS: Human VSMCs and male C57BL/6 mice were used for establishing AS cell models and animal models. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expressions of miR-135a-5p, forkhead box O1 (FOXO1) mRNA, and Janus kinase 2 (JAK2) mRNA. CCK-8, BrdU, and Transwell assays were used to detect cell migration and proliferation. Cell cycle and apoptosis were analyzed using flow cytometry. The interactions among miR-135a-5p, FOXO1 and JAK2 were validated employing Western blot, qRT-PCR and Luciferase reporter gene assay. RESULTS: The expression of miR-135a-5p was significantly decreased in serum samples of AS patients, VSMCs treated with ox-LDL and AS mice models. The overexpression of miR-135a-5p induced VSMCs cycle arrest and apoptosis, and inhibited proliferation and migration. Further experiments confirmed that miR-135a-5p could target and repress FOXO1/CyclinD1 and JAK2/STAT3 pathway. Additionally, the associations among miR-135a-5p, FOXO1/Cyclin D1 and JAK2/STAT3 were validated using animal models. CONCLUSION: MiR-135a-5p suppresses VSMCs proliferation and migration induced by ox-LDL via targeting and activating FOXO1/Cyclin D1 and JAK2/STAT3 signaling pathways.