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目的探索miR-590-5p在A375恶性黑素瘤细胞中的表达及其对A375细胞侵袭和迁移能力的影响和机制。方法应用实时定量PCR技术检测A375细胞与人正常黑素细胞中miR-590-5p的表达水平;应用TranswellTM实验和划痕实验检测miR-590-5p对A375细胞侵袭和迁移的影响;应用荧光素酶报告基因实验明确yes相关蛋白(YAP1)是否为miR-590-5p的直接靶分子;应用实时定量PCR及Western blot法检测miR-590-5p对YAP1表达水平的影响。结果 miR-590-5p在A375细胞中的表达水平显著低于正常黑素细胞;与对照(miR-590-5p-NC)组相比,miR-590-5p mimcs组细胞侵袭和迁移能力均受到显著抑制;YAP1为miR-590-5p的直接靶分子;与miR-590-5p-NC组相比,miR-590-5p mimcs组细胞中YAP1 mRNA及蛋白的表达均受到抑制。结论 miR-590-5p在A375细胞中低表达,并通过靶向抑制YAP1的表达调控A375细胞的侵袭和迁移能力。
Objective To explore the expression of miR-590-5p in A375 malignant melanoma cells and its effect on the invasion and migration of A375 cells. Methods Real-time quantitative PCR was used to detect the expression of miR-590-5p in A375 cells and human normal melanocytes. The effects of miR-590-5p on invasion and migration of A375 cells were detected by TranswellTM assay and scratch assay. Fluorescein Enzyme reporter assay confirmed whether yes-related protein (YAP1) is a direct target of miR-590-5p. The effect of miR-590-5p on the expression of YAP1 was detected by real-time PCR and Western blot. Results The miR-590-5p expression level in A375 cells was significantly lower than that in normal melanocytes. Compared with miR-590-5p-NC group, miR-590-5p mimcs cells were affected by invasion and migration YAP1 was a direct target of miR-590-5p. Compared with miR-590-5p-NC group, the expression of YAP1 mRNA and protein in miR-590-5p mimcs group was inhibited. Conclusion miR-590-5p is down-regulated in A375 cells and regulates the invasion and migration of A375 cells by targeting YAP1 expression.