Objective To investigate the effect of long-wave ultraviolet on the expression of opsin 3 in human melanoma cells lines(A375 cells and MV3 cells)and its impact on cell proliferation,as well as to explore the molecular mechanism of opsin 3 regulation on human melanoma cell proliferation.Methods Human melanoma cell lines(A375 cells and MV3 cells)were exposed to various doses of UVA,and cell proliferation was assessed by the 5-ethynyl-2-deoxyuridine(EdU)and Cell Counting Kit-8(CCK-8)experiments.The transcription levels of opsin in human melanoma cell lines(A375 cells and MV3 cells)were assessed by real time quantitative PCR(RT-qPCR)and Western blot.Lentivirus infection were utilized to silence or overexpress opsin 3 in A375 cells and MV3 cells,with subsequent assessment of cell proliferation levels by the EdU method and CCK-8 assay.After A375 cells and opsin 3 in MV3 cells were silenced,the changes of cell proliferation before and after ultraviolet A(UVA)irradiation by EdU method and CCK-8 assay.RNA-seq was used to detect differential gene expression in OPN3-overexpressing or silenced MV3 cells,with subsequent analysis conducted on KEGG enrichment-related signaling pathways.Western blot was used to verify purposes regarding the expression of Hippo signaling pathway-related proteins.Results Compared with the control group,the cell proliferation ability of UVA group was significantly enhanced,and this difference was statistically significant(P＜0.05).RT-qPCR revealed that opsins 1,2,3,4,and 5 were expressed in A375 and MV3 cells.The transcriptional expression and protein expression of opsin 3 were significantly higher than those of other opsins(P＜0.05).Long wave ultraviolet irradiation significantly increased the expression of opsin 3 protein in A375 and MV3 cells.With the silencing of opsin 3,the proliferation ability was weakened while overexpression enhanced it(P＜0.05).The proliferation ability of A375 cells and MV3 cells after the silence of OPN3 by UVA irradiation was significantly attenuated compared with that by UVA irradiation alone.RNA-seq screened out 141 differential genes upon OPN3 silencing or over-expression in MV3 cells.KEGG analysis further revealed 9 enrichment signaling pathways of differential genes with Hippo signaling pathway as the highest-Log10(FDR)value.With the silencing of OPN3,large tumor suppressor 1(LATS1)expression levels associated with Hippo signaling pathway increased while Phospho-Yes-associated protein(p-YAP),Yes-associated protein(YAP),ras homolog family member A(RhoA)protein expression levels decreased.Conclusion UVA can promote the expression of opsins,specifically OPN3 in human melanoma cell lines(A375 cells and MV3 cells),thereby regulating cell proliferation through the Hippo signaling pathway.
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