Experimental Study On MiR-138 Inhibiting The Invasion Of Melanoma Cell Invasion By Targeted Downregulation Of Endoglin

ObjectiveThis study analyzed the effect of miR-138 targeting endoglin in inhibiting the invasion of melanoma cells through in vitro cell experiments and animal models,and studied the related mechanisms in order to clarify the mechanism of melanoma occurrence and development.The development and application of targeted therapeutic drugs lay a theoretical and experimental foundation.MethodsThe melanoma cell lines WM451,WM266-4,SK-MEL-2,A375 and normal epidermal melanoma cell lines HEMA-LP were cultured,and the expression level of miR-138 was detected by fluorescence quantitative PCR to screen out the melanoma cell lines suitable for this study.WM451 cells were divided into groups and transfected with negative control(NC)mimic,miR-138 mimic,NC si RNA,endoglin si RNA,pc DNA3.1 plasmid and pc DNA3.1-endoglin plasmid,respectively.The expression levels of miR-138 and endoglin m RNA in each group were detected by fluorescence quantitative PCR,and the expression levels of endoglin protein were detected by Western blot.The effects of miR-138 and endoglin on the invasion of melanoma cells were detected by Transwell assay.Bioinformatics method was used to predict the target genes of miR-138,and dual luciferase reporter genes were used to verify the prediction results.WM451 cells stably transfected with negative control lentivirus or miR-138 lentivirus were injected into mice through tail vein to observe the number of tumor nodules in the lungs of mice.ResultsThe expression level of miR-138 in WM451,WM266-4,SK-MEL-2 and A375 cells was lower than that in HEMA-LP cells.Among all the melanoma cells,the expression level of miR-138 in WM451 cells was the lowest,so WM451 cells were used for subsequent experimental studies.When the expression of miR-138 was upregulated,the invasion ability of WM451 cells was significantly inhibited.Endoglin was predicted to be a target gene of miR-138.The luciferase reporter gene system confirmed that the fluorescence activity of WM451 cells transfected with wild-type endoglin double luciferase reporter gene plasmid was significantly lower than that transfected with mutant endoglin.When the expression level of miR-138 was upregulated,the expression levels of endoglin m RNA and protein in WM451 cells were significantly inhibited.The invasion number of WM451 cells in si-endoglin group was lower than that in si-NC group.The invasion number of WM451 cells in the pc DNA3.1-endoglin+miR-138 group was higher than that in the pc DNA3.1+miR-138 group.Mice injected with WM451 cells upregulated with miR-138 showed fewer lung tumor nodules than those in the NC group.ConclusionsmiR-138 can inhibit the invasion of melanoma cells,and targeting endoglin is one of the molecular mechanisms that mediate this inhibition.