The Anti-tumor Effects Of DC-CIK Co-cultured Cells Combination With Sorafenib Against Hepatocellular Carcinoma In Vitro And In Vivo

ObjectiveTo investigate the inhibitory effects of DC and CIK co-cultured cell(DC-CIK) combined with sorafenib on the expansion of Hepatocellular carcinoma cell line BEL-7402 in vitro and in vivo.Methods一. DC and CIK culture in vitroDC and CIK cells were generated in vitro by stimulation of peripheral blood mononuclear cells subsets with different cytokines and co-cultured on the eighth day. Phenotype of DC and CIK cells was detected by flow cytometry.二. specific kill effect and apoptosis rate detectionThe specific cytotoxicities of DC-CIK combined with sorafenib against Hepatocellular carcinoma cell line BEL-7402 was determined by Cell Counting Kit-8. The Annexin V-FITC Apoptosis Derection Kit was used to detect apoptosis. DNA nuclear morphology was examined with Hoechst 33258.三. Nude mouse xenograft assayHepatocellular carcinoma cell line BEL-7402 were injected subcutaneously into nude mice. On the 14th day,normal saline (control group),sorafenib group,DC-CIK group and combination group were administered respectively. Tumor volume was measured after treatments and tumor inhibitory was calculated.Results一. DC and CIK culture in vitro After adding tumor cell lysate and cytokine, the mature markers of DC increase, such as CD83, CD80, CD86, HLA-DR. CD3+/CD56+cell is the main effector cell of CIK cells,According to the total number of CIK cell proliferation the absolute counting of CD3+/CD56+cell was increased by over 1000 times.二. specific kill effect and apoptosis rate detectionThe results of CCK-8 demonstrated that the combination of DC-CIK and sorafenib possessed more obvious Cytotoxic activity to BEL-7402 cells.DC-CIK combined with sorafenib can improve the inhibition rate and also promote apoptosis(P<0.05).三. In vivo antitumor effect of DC-CIK and sorafenibThe antitumor activity in nude mouse xenograft assays was evaluated. DC-CIK combined with sorafenib inhibited 83.37% of BEL-7402 tumor growth in nude mouse xenograft assays. Volume of implanted tumor in combination group was significantly smaller than in the other groups(P<0.05).ConclusionDC-CIK cells combined with sorafenib can inhibit the growth of Hepatocellular carcinoma cell line BEL-7402 in vitro and in vivo. Molecular targeting therapy combined with cytoimmunity therapy may be a new way for the comprehensive treatment of Hepatocellular carcinoma.