Study on the mechanism of miR-141-5p affecting the biological behavior of mucosal melanoma cells
Objective To explore the expression of miR-141-5p and its significance in mucosal melanoma(MM)tissues and A375 cells,and to analyze the mechanism by which it affects the biological behavior of MM cells.Methods MM and pigmented nevus specimens surgically excised from the First Affiliated Hospital of Wenzhou Medical University were collected from January 2015 to December 2020.qRT-PCR was performed to detect the expression levels of miR-141-5p,and extracellular signal-regulated kinase(ERK)1/2 mRNA in MM tissues and pigmented nevus tissue,MM cell line A375 and human keratinocyte cell line HaCaT,and Pearson correlation analysis was used to anaiyze the correlation between miR-141-5p and the relative expression of ERK1/2 mRNA.A375 was treated with miR-141-5p mimic(mimic group),miR-141-5p mimic blank sequence(mimic control group),miR-141-5p inhibitor(inhibitor group),and miR-141-5p inhibitor blank sequence(inhibitor control group),respectively;and qRT-PCR was performed to detect the expression levels of miR-141-5p in the cells of the four groups;Cell counting kit(CCK)-8 method,clone formation assay,flow cytometry,scratch assay and Transwell invasion assay were used to observe and compare the survival rate,proliferation ability,apoptosis rate and cell migration and invasion ability of the four groups of cells;Western blot was used to observe and compare the relative expressions of ERK1/2 and phosphorylated ERK1/2(p-ERK1/2)proteins in A375 cells of the four groups.HE staining and immunohistochemical staining were used to observe the morphology and detect the expressions of p-ERK1/2 and ERK1/2.The differences in p-ERK1/2 and ERK1/2 expression between MM patients with different clinicopathological characteristics were compared,respectively.Results miR-141-5p and ERK1/2 mRNA expression were down-regulated in MM tissues and A375 cells(all P＜0.01);Pearson correlation analysis showed that miR-141-5p was positively correlated with the expression level of ERK1/2 mRNA(P＜0.05).Compared with the control group,the cell proliferation and migration and invasion abilities were significantly reduced and apoptosis rate was elevated in the mimic group,while the cell proliferation and migration and invasion abilities were elevated and apoptosis rate was reduced in the inhibitor group(all P＜0.05).Compared with the control group,the relative expression levels of p-ERK1/2 proteins in the mimic group were significantly increased,while significantly lowered in the inhibitor group(all P＜0.05).The expressions of ERK1/2 and p-ERK1/2 proteins in MM tissues were significantly reduced(both P＜0.05).There were no significant differences in clinicopathological features between p-ERK1/2 negative group and positive group or ERK1/2 negative group and positive group(all P＞0.05).Conclusion miR-141-5p may inhibit the biological behavior of MM cells through the ERK1/2 pathway.
万方数据
维普
