RESUMO
Objective To detect the expression of miR-590-5p in malignant melanoma A375 cells, its effect on the invasion and migration of A375 cells, and underlying mechanism. Methods Real-time PCR was performed to detect the expression of miR-590-5p in A375 cells and human normal melanocyte (HM) cells; TranswellTM assay and wound healing assay were used to observe the effect of miR-590-5p on the invasion and migration of A375 cells; Luciferase assay was done to determine whether YAP1 was the direct target of miR-590-5p; quantitative real-time PCR and Western blotting were performed to investigate the effect of miR-590-5p on YAP1 expression. Results Compared with HM cells, miR-590-5p was significantly down-regulated in A375 cells. Compared with A375 cells transfected with miR-590-5p-NC, cell invasion and migration were significantly inhibited in A375 cells transfected with miR-590-5p mimics. Luciferase assay indicated that YAP1 was the direct target of miR-590-5p. Compared with A375 cells transfected with miR-590-5p-NC, mRNA and protein levels of YAP1 were significantly inhibited in A375 cells transfected with miR-590-5p mimics. Conclusion miR-590-5p is downregulated in A375 cells, and inhibits the invasion and migration of A375 cells by directly regulating the expression of YAP1.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Melanoma/genética , Melanoma/fisiopatologia , MicroRNAs/metabolismo , Fosfoproteínas/genética , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Humanos , Melanoma/metabolismo , Melanoma/patologia , MicroRNAs/genética , Invasividade Neoplásica , Fosfoproteínas/metabolismo , Fatores de Transcrição , Proteínas de Sinalização YAPRESUMO
OBJECTIVE: To find herbs with effects on adhesion and migration of melanocytes in vitro. MATERIALS AND METHODS: Ethanol extracts from 14 herbs were tested. Normal human melanocytes were obtained from neonatal foreskin, and the 48-well culture dish covered with fibronectin was used for the melanocyte adhesion assay. Motility was assessed by using the micropore filter method. RESULTS: The extracts of Danshen (Radix Salviae Miltiorrhizae), Tusizi (Semen Cuscutae) and Honghua (Flos Carthami) could enhance melanocyte adhesion to fibronectin, while Cijili (Fructus Tribuli) and Huangqi (Radix Astragali) promote melanocyte migration in vitro. Buguzhi (Fructus Psoraleae), Nü Zhen Zi (Fructus Ligustri Lucidi) and Baizhi (Radix Angelicae Dahuricae) could promote both adhesion and migration of melanocytes. CONCLUSION: The above herbs may play a role through promoting adhesion and/or migration of melanocytes in the treatment of vitiligo.