Profiling changes in dendritic cells pulsed by hepatocellular carcinomas with different metastatic potentials

Abstract

Objective To explore the mechanism of HCC immune escape, we utilized proteomic approach to analyze the relations between expression profiles changes of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) lysates of different metastatic potentials and the changes of DC's function. Methods DCs were derived from human peripheral blood monocytes (PBMC) and pulsed with HCC lysates of different metastatic potentials. The normal liver cell line lysate-pulsed DCs and norrlysate pulsed DCs were employed to serve as the control. The total protein of these DCs was identified by immobilized pH gradient two-dimensional poly-acrylamide gel electrophoresis (2DE), silver stain, Pdquest 2DE analysis software, ESI, and IPI database search to yield the different profiles among them. Results The match ratio of all samples' 2DE was higher than 80% while the correlation coefficient over 0. 8. There were 21 spots exhibiting quantitative changes that more than 3 folds among DCs pulsed with different lysates, 12 of them were identified by ESI. These proteins were related to cytoskeleton ( spot 3: Keratin type 1 cytoskeletal 10 ; spot 16: TPMsK3; and spot 21: p-centractin), signal transduction ( spot 8: Cadherin-related tumor suppressor homolog precersor; spot 11 Annexin A2 Iso-form 2; spot 17: Late endosomal/Lysosomal Mp 1 interacting precursor), cytoplasmic movement and energic metabolism ( spot 6:Ubiquinol-cytochrome C reductase complex core protein I, mitochondrial precursor;spot 9: Biliverdin reductase A precersor ; spot 13: Cytosolic malate dehydrogenase ; and spot 20: Methylmalonate-semialdehyde dehydrogenase [cylating], mitochondrial precursor) and others ( spot 10: Anti-colorectal carcinoma heavy chain ;spot 18: Transgelin-2 ). Conclusion These 12 spots represent proteins of cytoskeleton, signal transduction as well as proteins involved in cytoplasmic movement and energic metabolism. Some of them might be related to the function of DCs and metastatic potential of HCC. Key words: Carcinoma, hepatocellular; Dendritic cell; PAGE-2DE; ESI