Screening and identification of suppressor proteins targeting dendritic cells in peripheral blood of hepatocellular carcinoma
Objective To screen and identify the inhibitory proteins targeting dendritic cells in peripheral blood of hepatocellu-lar carcinoma.Methods CD14+Monocyte from healthy human peripheral blood were sorted and induced to differentiate into immature dendritic cells(iDCs),and then into mature dendritic cells(mDCs).The purity of iDCs was identified.Enzymatic digestion of binding proteins of iDCs incubated with serum from healthy individuals and liver cancer patients,recovery of peptides after enzymatic digestion,and screening of potential interacting candidate proteins that have potential regulatory effects on the differentiation and maturation of iDCs.Targeted identification of differential candidate proteins.The overexpression protein particles were constructed,transfected into nor-mal hepatocytes(HL7702)and cultured.The supernatant of overexpression protein hepatocytes and normal hepatocytes were co cultured with iDCs to induce the maturation of iDCs.The hepatoma derived serum proteins that specifically inhibit the maturation of iDCs were screened and identified.Results The proportion of pre induction antigen marker positive cells conforms to the antigen phenotype charac-teristics of iDCs;The proportion of antigen marker positive cells after induction conforms to the antigen indication characteristics of mDCs.Preliminary identification of over 40 potential interacting candidate proteins that have potential regulatory effects on the differentia-tion and maturation of iDCs.Select 10 differential candidate proteins.Verification of its overexpression showed that overexpression of AFP protein,AGXT protein,and ATP1A1 protein significantly inhibited the differentiation and maturation of iDCs into mDCs.Conclusion Ten inhibitory proteins targeting dendritic cells in peripheral blood of hepatocellular carcinoma were screened.AFP,AGXT and ATP1A1 could significantly inhibit the maturation of dendritic cells.
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