Research For Effect Of CIK Cells Combined With Dendritic Cells On The Cycle, Poptosis, Proliferation And Migration Of Hepatoma Carcinoma Cells

Objective: To explorate the effect of cytokine induced killer (CIK）celland dendritic cell（DC）from peripheral blood mononuclear cell (PBMC) ofpatient with primary liver cancer on the cycle, apoptosis, proliferation andmigration of human HEPG2cell line of liver cancer.Methods: The PBMC were induced by GM-CSF、TNF-αand IL-4inorder to produce DC, which were sensitized with antigen of autologoustissue of primary liver cancer. Meanwhile, CIK cells were obtained from thesuspended PBMC induced by IFN-γ、IL-2and human CD3monoclonalantibody（hCD3mAb). The DC, CIK cells and HEPG2cell line werecultured in a same circumstance but different parts of a culturing cabinnamed Transwell for24h and48h respectively. Subsequently the cycle ofhuman HEPG2cell line were tested by flow cytometry (FCM). The AnnexinV-PI staining was employed to analysis the apoptosis of the same cell line.The expression of Bax gene which related with apoptosis of this cells and itscoded protein were inspected with the methods of RT-PCR and Western blot. The proliferations of human HEPG2cell line were determined with cellcount kit-8(CCK-8), and the proliferation curve was made. The scratch testwas employed to analysis the proliferation and migration of the same cellline. The expression of PCNA gene related closely proliferation of this cellsand its coded protein were inspected with reverse transcriptase-polymerasechain reaction (RT-PCR) and Western blot respectively.Results: DC-CIK cells can markedly influence the cycle and theproliferation of HEPG2cell line，noticeably block it in the phase of G0andinduce its apoptosis (compared with control, P<0.01). The results of CCK-8shows DC-CIK cells can markedly influence the proliferation of HEPG2cellline (compared with control, P<0.01). The tests of scratch, gene and proteinindicated DC-CIK cells can significantly inhibit proliferation and migrationof human HEPG2cell line and down-regulate the expression of PCNA generelated closely the proliferation of the same cell line.Conclusions: DC-CIK cells of patient with primary liver cancer canobviously inhibit the growth of human HEPG2cell line, down-regulate theexpression of PCNA gene of human HEPG2cell line, inhibit its proliferationand migration and noticeably induce its apoptosis which posses thecharacteristics of time-dependence and cell population-dependence.