Mechanism research of PAD4-mediated H3cit to promotes the proliferation,invasion and epidermal-mesenchymal transition of colon cancer cells
Objective To explore the role and mechanism of peptidyl arginine deiminase 4(PAD4)-mediated citrullinated histones H3(H3cit)in the proliferation,invasion and epidermal-mesenchymal transition of colon cancer cells.Methods Serum samples were collected from 30 colon cancer patients and 30 healthy subjects in our hospital.The levels of PAD4 and H3cit in serum were detected by ELISA,and the correlation between the two was analyzed.oe-NC,overexpressed PAD4(oe-PAD4),sh-NC and sh-NLRP3 vectors were transfected into human colon cancer SW480 cells and divided into the oe-NC group,the oe-PAD4 group,the oe-PAD4+sh-NC group,and the oe-PAD4+ sh-NLRP3 group.The gene expression levels of PAD4,NLRP3,IL-1β and IL-18 were detected by qRT-PCR,the protein expression of PAD4,H3cit,NLRP3,E-cadherin and N-cadherin were detected by Western blot.The cell proliferation level was detected by CCK-8,and the level of cell invasion was detected by Transwell assay.The level of H3cit enrichment in NLRP3 transcription start sites(TSS)was detected by ChIP-qPCR.Results Compared with healthy subjects,the levels of PAD4 and H3cit in serum of colon cancer patients were increased,and there was a positive correlation between the two(P<0.05).Compared with the oe-NC group,the gene expression levels of PAD4,NLRP3,IL-1β and IL-18 were increased,the protein expression of PAD4,H3cit,NLRP3 and N-cadherin were increased,the protein expression of E-cadherin was decreased,the cell proliferation and invasion levels were increased,and the enrichment of H3cit in NLRP3 TSS zone was increased in the oe-PAD4 group(P<0.05).Compared with the oe-PAD4+sh-NC group,the gene expression levels of NLRP3,IL-1β and IL-18 were decreased,the protein expression of NLRP3 and N-cadherin were decreased,the protein expression of E-cadherin was increased,and the cell proliferation and invasion levels were decreased in the oe-PAD4+sh-NLRP3 group(P<0.05).Conclusion The levels of PAD4 and H3cit in colon patients are increased,and the PAD4-mediated H3cit promotes the proliferation,invasion and epithelial-mesenchymal transition of colon cancer cells by transcriptively regulating the expression of NLRP3.
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维普
