Bioactivity Of Cytokine-Induced Killer Cells On Hepatocellular Carcinoma Cells In Vitro

Objective To seperate mononuclear cells from healthy volunteers' peripheral bloo d, the cells come into CIK cells after induced by cytokines and anti-CD3 antibodies,Analyze the phenotype of CIK cells.Method Extract peripheral blood from healthy volunteers' ulnar vein, density gra dient centrifugation, peripheral blood monocyte(PBMC), To grow the cells into culture bottle packaged by CD3 monoclonal antibody, then put IFN-??IL-1? and IL-2 into t he bottle. Analyze the phenotype of and CIK cells in different days.Results The quantity of CIK cells were very few at the first 3 days, the quantit y of cells become fold increase at the 6 day, the cell proliferate ability riches its pea k at the 15 th day, The rate of CD3CD56 CIK cells goes up with the incubation time.Conclusion Our way of cultural CIK cell method is reliable, the CIK cells have high activity and multiplicate largely.Obiective Culture the CIK cells together with 3 hepatocellular carcinoma(HCC) cells. Test the apoptosis and colon formation of 3 hepatocellular carcinoma(HCC) cell lines in votro.Methods The CIK cells were cultured with 3 kinds of HCC cell lines(Hep G2, Hep3 B and Huh7 cells) in the same circumstance for 24 h and 48 h, respectively. And then the proliferation of these cancer cells was determined with cell count kit-8(CCK-8). The cell colon fomation was analysised by agrose cloning formation asssay and Annexin V-PI double staining was employed to analysis the apoptosis of cancer cell lines. m RNA and protein expression of Bax and Caspase-3 was determined by reverse transcriptase-polumerase chain reaction(RT-PCR) and Western blotting, respectively.Results The results of CCK-8 showed that CIK cells markedly inhibited the proliferation of cancer cell lines(compared with control, P<0.01). The results of Annexin V-PI staining showed that CIK cells increased the apoptosis of cancer cells(increased proportion of apoptotic HCC cells, and elevated expression of Bax and Caspase-3) and the effects of CIK on HEPG2 cell line were stronger than the other 2 cancer cell lines.Conclusion CIK cells from healthy volunteers with can obviously inhibit the growth of cancer cells, inhibit their proliferation and induce their apoptosis in a time-dependent way.