Mechanism Of DSG2 Gene In Etoposide-induced DNA Damage Signaling Pathway In Cervical Cancer Hela Cells

Objective: In order to understand the mechanism of Desmoglein2(DSG2)gene signaling pathway involved in DNA damage of cervical cancer Hela cells,and to screen the molecular targets related to DNA damage of cervical cancer,it provides a new research direction for clinical diagnosis and treatment of cervical cancer.Methods:(1)The expression of DSG2 protein in cervical carcinoma and paracancerous tissue microarray was detected by immunohistochemistry;(2)Western blot showed that the overexpression of DSG2 was significantly increased in the overexpressed Hela cell line,and the expression of DSG2 was significantly decreased in the knocked out Hela Cell Line;(3)DNA damage was induced by Etoposide in three groups of DSG2 overexpression,knock-out and vector Hela cell lines,and then transcribed separately to obtain differentially expressed genes.The genes were enriched by GO and KEGG,and the corresponding results were obtained;(4)The protein-protein Interaction(PPI)network was plotted using the STRING database and Cytoscape 3.6.1 software to identify key genes.Results:(1)Immunohistochemical results showed that DSG2 expression was significantly increased in cervical cancer compared with adjacent tissues(P<0.05);(2)After constructing a DSG2 overexpressed and knocked out Hela cell line with CRISPR/Cas9,Western blot showed that the overexpression of DSG2 was significantly increased in the overexpressed Hela cell line,and the expression of DSG2 was decreased considerably in the knocked out Hela Cell Line(P<0.05);(3)Hela cells with vector were induced by Etoposide and Dimethyl Sulfoxide respectively.Compared with the DMSO group,Etoposide induced vector Hela cells had 960 differentially expressed genes,including 374 upregulated genes and 586 downregulated genes.After the differentially expressed genes were enriched in the KEGG database,the p53 signal pathway and cancer-related signal pathway were the most abundant;(4)DSG2 overexpressed,knocked out,and vector Hela cell lines were all induced by sequenced,respectively.Compared with vector Hela cell line,DSG2 overexpression cell line had 6,514 differentially expressed genes,of which 3,260 genes were upregulated and 3,254 genes downregulated;Compared with vector Hela cell lines,DSG2 knocked out cell lines had 5,444 differentially expressed genes,of which2,828 genes were upregulated and 2,616 genes downregulated;(5)The enrichment of the differentially expressed gene GO in the two groups with DSG2 overexpression and knocked out showed that the differentially expressed genes could be enriched in both intracellular part and intracellular part,and the binding in the molecular function.KEGG results showed that the differentially expressed genes were mainly enriched in the ribosomal signaling pathway;(6)A visual PPI network based on Dsg2 overexpression Group differentially expressed genes.Among them,ten upregulated hub genes such as RPS23 and RPS19 and ten downregulated hub genes such as PSMB8 and HLA-A are the central targets of the network.Conclusion:(1)DSG2 gene is overexpressed in cervical cancer and may be a potential tumor marker of Cervical Cancer;(2)KEGG and PPI analysis showed that Etoposide induced DGS2 gene overexpression in the cell.In the meantime,RPS23 and RPS19 genes are upregulated in the ribosomal pathway,promoting DNA damage in tumor cells.