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Huadong Liu, Wei Xiong, Qiyun Liu, Jian Zhang, Shaohong Dong
(Department of Cardiovascular Medicine, The Second Clinical College of Jinan University, Shenzhen People’s Hospital, Shenzhen, Guangdong, China (mainland))
Med Sci Monit 2016; 22:4054-4061
We aimed to explore how chemokine-like receptor 1 (CMKLR1) influences the proliferation and migration of vascular smooth muscle cells (VSMCs).
MATERIAL AND METHODS: Normal VSMCs, negative control VSMCs interfered by CMKLR1 gene, and VSMCs with stable knockdown of CMKLR1 gene were divided into the control group, PDGF group, negative-shRNA group, and CMKLR1-shRNA group. Both cell number counting and BrdU incorporation assays were employed to investigate the proliferation status of VSMCs. Transwell migration assay was used to measure the migration status of VSMCs. Inflammation markers, including cytokines IL-1β, IL-6, TNF-α, and chemokines MCP-1 in VSMCs, were detected by real-time quantitative RT-PCR. Western blotting assay was used to detect protein expressions of the MAPK pathway in VSMCs.
RESULTS: The number of VSMCs and the OD value of BrdU in PDGF group were significantly higher than those in the control group (both P<0.05). Compared with the control and negative-shRNA group, the CMKLR1-shRNA group exhibited significantly reduced VSMCs number and BrdU OD value (both P<0.05). Transwell migration assay indicated that PDGF-BB promoted whereas CMKLR1-shRNA inhibited the migration of VSMCs. The expression of IL-1β, IL-6, TNF-α, and MCP-1 were up-regulated in the PDGF group but down-regulated in the CMKLR1-shRNA group. Compared with normal VSMCs, the protein level of p-ERK1/2 was up-regulated in VSMCs treated with PDGF-BB, while it was down-regulated in the CMKLR1-shRNA group.
CONCLUSIONS: CMKLR1 exacerbated the proliferation and migration of VSMCs by activating ERK1/2.
Keywords: Cell Proliferation - physiology, Cell Movement - physiology, Animals, Cells, Cultured, Chemokines - metabolism, Cytokines - metabolism, Gene Expression, Gene Knockdown Techniques, MAP Kinase Signaling System, Mice, Muscle, Smooth, Vascular - metabolism, Phosphorylation, Receptors, Chemokine - genetics, Receptors, G-Protein-Coupled - genetics