Construction and functional study of vascular smooth muscle cells with extracellular matrix metalloproteinase inducer mutation
Objective To explore the effects of extracellular matrix metalloproteinase inducer(EMMPRIN)on the proliferation,migration and inflammatory response of vascular smooth muscle cells(VSMCs).Methods CRISPR/Cas9 technology was used to construct EMMPRIN c.889C>A-point mutation VSMC line.Real-time qPCR and Western blot were performed to detect the expression level of EMMPRIN.Cell proliferation activity was measured using the CCK-8 assay,while cell migration capability was evaluated using the Transwell assay.Secretion of interleukin-6(IL-6),monocyte chemoattractant protein-1(MCP-1),and interleukin-1β(IL-1β)was detected using enzyme-linked immunosorbent assay(ELISA).Results Compared to the wild-type(WT)cells,the expression level of EMMPRIN did not exhibit significant changes in c.889C>A mutant cells(t=0.732,P>0.05).There was no statistically significant difference in cell proliferation activity(P>0.05),while cell migration capacity significantly decreased(t=5.121,P<0.05).Secretion of inflammatory cytokines IL-6,MCP-1 and IL-1β induced by TGF-β decreased markedly in the c.889C>A mutant cells(t=9.371,5.690,6.192;P<0.05).Conclusion The EMMPRIN c.889C>A mutation does not affect the expression of EMMPRIN or VSMCs proliferation.However,it can supress the migration of VSMCs and the secretion of inflammatory cytokines.
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