Abstract
Objective To explore the effects and mechanism of cytokine induced killer (CIK)cells and dendritic cells (DC) from peripheral blood mononuclear cells (PBMC) of patients with human colorectal cancer on prloferation and apoptosis of human liver cancer cell line SW620.Methods Adherent PBMC were induced by granulocyte macrophage-colony stimulating factor (GM-CSF),tumor necrosis factor (TNF)-α and interleukin (IL)-4 to prepare DC.CIK cells were obtained from suspended PBMC induced by interferon (IFN)-γ,IL-2 and human CD3 monoclonal antibody.The colorectal cancer SW620 cells that were cultured without DC-CIK cells served as the controls.DC-CIK cells and SW620 cells were cultured in a well cell cultrue cluster for 24,48 and 72 h,respectively.The proliferation of SW620 cells that were cultured with DC-CIK cells for 24,48 and 72 h was detected by methyl thiazol tetrazolium (MTT) assay.The telomerase activity was determined by tartrate resistant acid phosphatase (TRAP)-enzyme linked immunosorbent assay (ELISA).Human telomerase reverse transcriptase (hTERT) gene was evaluated by Western blotting.Results In colorectal cancer cells treated with DC-CIK after 72 h treatment,cell relative growth rate in control group,and DC-CIK-a group and DC-CIK-b group was (95.6 ± 2.2) ％,(73.5 ±1.8)％ and (51.8 ± 1.6)％ respectively.The results from TRAP-ELISA showed that telomerase activity in control group at 24,48 and 72 h was 1.568 ±0.078,1.535 ±0.056,and 1.527 ±0.039,that in DCCIK-a group was 1.356 ±0.056,1.108 ±0.039,and 0.578 ±0.028,and that in DC-CIK-b was 1.181 ±0.075,0.718 ± 0.039,and 0.386 ± 0.036,respectively.The Western blotting results showed that the hTERT protein in DC-CIK-a group and DC-CIK-b group was down-regulated as compared with control group.Conclusion DC-CIK cells could inhibit the proliferation of colon cancer cells through down-regulating telomerase activity. Key words: Colorectal cancer; Dendritic cell-cytokine induced killer; Proliferation; Telomerase
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