Regulation Of LncRNA-177922 On Melanogenesis,Proliferation,Migration,and Autophagy Of Melanoma Cells B16-F10 By Targeting MAPK15

LncRNAs play important roles in the life activities of normal tissue cells and tumors.The differentially expressed LncRNA-177922 was obtained,based on the the previous transcriptome data analysis of C57/BL6 J mouse melanocytes and melanoma cells B16-F10.Mitogen-activated protein kinase15(MAPK15)is predicted to be the target gene of LncRNA-177922,which regulates the occurrence and development of tumors.Therefore,LncRNA-177922 may be related to melanoma cell biology.In order to investigate the role of LncRNA-177922 in melanoma cells,LncRNA-177922 was overexpressed in B16-F10 cells to analyze the biological changes and related molecular mechanisms of B16-F10 cells.The result were as follows:1.MAPK15-pmir GL0,LncRNA-177922-pc DNA3.1 or empty plasmid were co-transfected into 293 T cells.The results of dual luciferase activity showed that compared with the control group,the luciferase activity of LncRNA-1779252 and MAPK15 were decreased with significant different(P < 0.001).2.The effects of LncRNA-177922 on melanin production in B16-F10 cells were detected by q PCR,Western blot,Fontana-Masson staining.The results showed that LncRNA-177922 upregulated tyrosinase activity,and the expression levels of MITF and downstream TYR,TYRP1,and TYRP2 were increaseed,as a result of the increase of melanins production with significant different(P < 0.01).3.The related effects of LncRNA-177922 on cell proliferation of B16-F10 cells was detected by q PCR,Western blot,and CCK-8 assays.The results showed that LncRNA-177922 upregulated the expression levels of MEK1,ERK,and MNK1,resulting in proliferation with significant different compared with control cells(P < 0.01).4.The effect of LncRNA-177922 on the migration of B16-F10 cells was detected by Western blot and scratch healing test.The results showed that compared with the control group,LncRNA-177922 significantly up-regulated the expression levels of Rho and FAK proteins,promoting the migration of B16-F10 cells(P < 0.01).5.The effect of LncRNA-177922 on the autophagy of B16-F10 cells was detected by q PCR,Western blot and cell immunofluorescence.The results showed that compared with the control group,LncRNA-177922 down-regulated the expression level of LC3 B,while up-regulating m TOR PI3 K and AKT,which caused the inhibitor of autophagy in B16-F10 cells(P <0.01).In summary,the results suggested that LncRNA-177922 was involved in regulating the biological characteristics of B16-F10 cells such as melanogenesis,proliferation,migration,and autophagy by targeting MAPK15,which might be a new therapeutic target and diagnostic marker.