Screening and analysis of differentially expressed genes of human melanocytes in skin cells mixed culture.

This study aims to screen the key genes and possible signaling pathways involved in the differentiation and proliferation of human melanocytes (MCs) by in vitro culture of mixed skin cells. This will be helpful to further study the mechanisms and treatment strategies of pigment-related diseases such as vitiligo.Mixed skin cells were obtained by digesting and separating normal human foreskin tissues. Ribonucleic acid (RNA) was extracted from sorting cells and high-throughput transcriptome sequencing was performed at different culture time points. Differentially expressed genes (DEGs) were obtained by comparing the expression abundance of genes at different culture time points. Then the key genes and signaling pathways involved in the differentiation and proliferation of MCs were screened and verified by real-time quantitative polymerase chain reaction (qPCR) test.Twenty one DEGs were finally screened for further qPCR validation, mainly involved in 4 signaling pathways. The expressions of Wnt5A, Wnt5B, FZD2 and FZD3 in Wnt pathway were continuously up-regulated, and that of Wnt4 gene was continuously down-regulated, however, all the above hadn't been verified by qPCR. The expressions of COL5A2, COL6A3, ITGB1, ITGA4, ITGAV, AKT3, PIK3CD, PIK3R1 and PIK3R2 in phosphoinositide 3-kinase (PI3K) pathway were continuously up-regulated, of which PIK3CD, PIK3R2, COL5A2, ITGA4, ITGAV and AKT3 were verified by qPCR. PDGFB and GRB2 gene expressions were down-regulated in platelet-derived growth factor (PDGF) pathway, while PDGFRB was continuously up-regulated, of which PDGFB and PDGFRB were verified. The DHRS3, DHRS9, RDH10 and SDR16C5 genes in retinol metabolic pathway were continuously down-regulated and verified by qPCR.We suggested that Wnt5A gene in Wnt/β-catenin classical pathway, integrin combining with extracellular matrix through PI3K signaling pathway, retinoic acid catabolism-related genes could promote the differentiation and proliferation of MCs; however, PDGFB gene might have a negative regulatory effect on the growth of MCs.