Coexpression analysis of angiogenesis, proliferation, apoptosis, autophagy and SHH pathway genes involved in skin expansion

Abstract

Skin and soft tissue expansion is a widely used technique in plastic surgery. However, the regulatory mechanisms associated with cellular processes involved in skin expansion are not well elucidated. In the present study, we aimed at exploring the transcriptome changes associated with skin expansion and profiling the difference in gene expression between the skin tissue in the top of the dilator and the skin tissue in the side of the dilator. A mouse model of skin expansion was established and RNA sequencing (RNA-Seq) was performed on samples collected at different time points. Differential expression analysis was performed using the DESeq2 package while STEM was used for time series clustering profiling. The regulatory networks were established and the functions of sets of genes were analyzed. The mRNA expression levels of candidate genes were validated by the quantitative RT-PCR. Among the skin tissue in the top of the dilator and normal samples at days 1, 3, 7, 14 and 28, 53 commonly upregulated and 7 commonly downregulated genes were identified while among the skin tissue in the side of the dilator and normal samples, 98 downregulated and 255 upregulated genes were identified. Genes differentially expressed among the skin tissue in the top of the dilator and normal samples were involved in coagulation and proliferation-associated pathways while those among the skin tissue in the side of the dilator and normal samples were involved in the inflammation, immune response, and defense response. Among the skin tissue in the top of the dilator and the skin tissue in the side of the dilator samples, 161 were constantly upregulated while 27 were constantly downregulated; these genes were enriched in the biological processes of cell adhesion and regulation of cell proliferation (n = 11). Furthermore, we identified that SHH signaling genes formed a coexpression regulatory network with cellular proliferation, apoptosis, autophagy and angiogenesis-related genes in the expanded skin. In conclusion, our findings can promote research and understanding of the mechanism of skin expansion and will find application in plastic surgery.

Keywords: Angiogenesis; Apoptosis; Autophagy; Cell proliferation; Shh signaling; Skin expansion; Transcriptome.