Screening of differentially expressed transcription factors in keratinocytes with overexpression of EGFR induced by rhPTH(1-34)

Abstract

Objective Using transcriptome sequencing, we screened differentially expressed transcription factors (TFs) in keratinocyte (KC) with overexpression of the epidermal growth factor receptor (EGFR) induced by recombinant human parathyroid hormone (1-34) in order to investigate the mechanism of rhPTH(1-34) in the treatment of psoriasis. Methods Overexpression of EGFR in HaCaT cells was achieved by lentiviral transfection. Cells in rhPTH (1-34) group were incubated with 0.01 μmol/L of rhPTH (1-34), while the control group were treated with DMEM solution alone. After extraction of RNA, the quantity and quality of RNA were assessed using UV absorbance assay and denaturing agarose gel electrophoresis, respectively. The cDNA was synthesized and amplified by real-time quantitative PCR for accurate quantification of the effective concentration of the library. DESeq2 software was used to analyze the differentially expressed genes, and significance was defined as |log2foldchange|≥1 and P<0.05. Results A total of 12 455 genes were detected in the rhPTH (1-34) and control groups, with 421 genes in the rhPTH (1-34) group and 461 genes only in the control group. Of 185 differentially expressed genes between the rhPTH (1-34)-treated cells and the control group, 108 genes were up-regulated and 77 genes were down-regulated. Among the down-regulated genes, top five differentially expressed transcription factors were ZNF525, ZNF774, ZNF576, ZBTB42 and POU4F1 (P<0.05). ZNF774 and POU4F1 were mainly involved in the regulation of Notch2 and MAPK signaling pathways, possibly regulating KC proliferation. Conclusions Inhibition of KC proliferation by rhPTH (1-34) may be associated with down-regulation of ZNF525, ZNF774, ZNF576, ZBTB42, and POU4F1 expression. ZNF774 and POU4F1 may be associated with KC proliferation.

Keywords

• rhpth(1-34)
• egfr
• keratinocyte
• transcription factor