Objective: To investigate the in vitro and in vivo inhibitory effects of DC (dendritic cell)-CIK (cytokine-induced killer cell) co-cultured cells combined with sorafenib against hepatocellular carcinoma cell line BEL-7402. Methods: DC and CIK cells were generated in vitro by stimulating human peripheral blood mononuclear cells with different cytokines, and then they were co-cultured. The cytotoxicity of DC-CIK co-cultured cells (DC-CIK) combined with sorafenib against BEL-7402 cells was determined by CCK8 kit. The apoptosis of BEL-7402 cells was measured by Annexin V-FITC Kit. BEL-7402-implanted tumor model was established by subcutaneous injection in nude mouse. Tumor-bearing mice were divided into normal saline control group, sorafenib group, DC-CIK group and DC-CIK+sorafenib group. The inhibitory effects were observed in different groups. Results: The cytotoxicity rate of BEL-7402 cells in DC-CIK+sorafenib group was significantly higher than those in the other two groups, with cytotoxicity rate in DC-CIK+sorafenib group being (75.24±1.91)%, which was 1.8 times that in DC-CIK group and 2.1 times that in sorafenib group (P<0.01). The apoptosis rate of BEL-7402 cells in DC-CIK+sorafenib group was significantly higher than those in the sorafenib and DC-CIK groups, with the apoptosis rate in DC-CIK+sorafenib group being (78.32±2.54)% (P<0.05). The volume of tumor in the combination group was significantly smaller than those in the other groups (P<0.05). In vivo results showed that DC-CIK+sorafenib treatment significantly inhibited the growth of BEL-7402-implanted tumors, and the inhibitory rate was (83.37 ±0.16)%, which was significantly higher than those of the other groups (P<0.01). Conclusion:DC-CIK co-cultured 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 immunotherapy may be a new way for the comprehensive treatment of hepatocellular carcinoma.

Objective To observe whether hepatitis B vaccine enhance the treating effect of cyto-kine induced kill(CIK) cells on hepatitis B virus transgenic(HBV-Tg) mice. Methods The HBV-Tg mice were treated with CIK cells by peritoneal injection and hepatitis B vaccine by hypodermic injection. The HBV DNA level were tested by real-time PCR,T lymphocyte subgroup were detected by flow cytometry and the pathological diversify of hepatic tissue were observed by HE staining. Results The HBV DNA loading in peripheral blood of HBV-Tg mice decreased after CIK cells were treated and CD3~+ , CD4~+ and CD8~+ cells increased which were enhanced after CIK cells combined with hepatitis B vaccine. Conclusion Hepa-titis B vaccine enhanced the treating effect of CIK on HBV-Tg mice which may be implemented by increased the blood level of CD3~+, CD4~+ and CD8~+ cells, especially CD8~+ cells level.

Objective To investigate the biological effects of Wnt gene in kidney cancer Caki?2 cells. Methods The Wnt gene was silenced in kidney cancer Caki?2 cells by lentivirus vector. The cell proliferate ability of cells in each group were assayed by CCK?8 kit at different time points. The apoptosis of Caki?2 cells was observed after silencing Wnt gene by transmission electron microscope. The invasion ability of each group cells was tested using Transwell chambers. The genes expression changes of Wnt/β?catenin signaling pathway and apoptosis related gene were determined by realtime PCR. Results Compared with the other two groups,the cell proliferate ability of the cells after silencing Wntgene was suppressed,and the difference was statistically significant(P<0.05). Apoptosis increased significantly in shRNA+Caki?2+Wnt group cells with silencing of Wntgene, and apoptotic body appeared in these cells. In invasive experiment,the number of emigrated cells in shRNA+Caki?2+Wnt group were significantly lower than other groups(P<0.05). The expression of Wnt mRNA,β?catenin mRNA and Bcl?2 mRNA in shRNA+Caki?2+Wnt group cells was lower than other groups(P<0.05). Conclusion Silencing of Wnt gene of kidney cancer Caki?2 cells can affect the proliferation rate of the cells, promote the cell apoptosis,and inhibit the invasion ability,which provide certain theoretical basis for the research and development of new drugs and new therapeutic targets.

BACKGROUND: Tumor-adopted immunity and gene transduction technique are used to introduce tumor necrosis factor-α vector into carrier cells, which are then re-infused into the body so that cancer cells can be killed by tumor necrosis factor-α more directly and effectively with fewer side effects on the other tissues due to high local expression.OBJECTIVE: To study the bioactivity of in vitro cultured tumor necrosis factor-α transduced tumor-infiltrating lymphocytes as well as the inhibitory effects on cancer cells of cancer-loaded rats infused in different ways.DESIGN: A randomized controlled study based on experimental animals.SEETING: Cancer Research Institute of China Medical University.MATERIALS: This study was carried out at the Cancer Research Institute and the Experimental Animal Department, China Medical University,between January 2000 and December 2001. TJ8510 cell line (human brain glioblastoma cell line) was provided by the Neurological Research Institute of Tianjin Medical University Affiliated Hospital. The experimental animals were 36 BALB/C nude mice congenitally having no thymius.METHODS: Based on the establishment of tumor necrosis factor-α retroviral transduction system and the preparation of cartier cells tumor-infil-trating lymphocytes, the monoclonal virus cell line PLC-2 and PLJC-5available were used to introduce marked gene NeoR and targeted gene tumor necrosis factor-α into tumor-infiltrating lymphocytes, respectively.Then cell proliferation, tumor necrosis factor expression and in vitro antitumor activity were examined. After cancer cell inoculation, the 36 nude mice were randomly divided into 6 groups: local infusion control group, local tumor-infiltrating lymphocytes infusion group, local tumor necrosis factor-tumor-infiltrating lymphocytes infusion group, venous infusion control group, venous tumor-infiltrating lymphocytes infusion group and venous tumor necrosis factor-tumor-infiltrating lymphocytes infusion group, and the therapeutic effects on the cancer-loaded mice were observed.proliferation and tumor necrosis factor-α expression in tumor-infiltrating oR-tumor-infiltrating lymphocytes and tumor necrosis factor-tumor-infiltrating lymphocytes was not significantly different from each other (P ＞ 0.05).NeoR-tumor-infiltrating lymphocytes, though not significantly different (P ＞0.05), significantly differ from that of tumor necrosis factor-tumor-infiltrating lymphocytes (P ＜ 0.01); moreover, tumor necrosis factor-tumor-infiltrating lymphocytes were found to express higher tumor necrosis factor-α conactivity did not significantly differ between tumor-infiltrating lymphocytes and NeoR-tumor-infiltrating lymphocytes (P ＞ 0.05), but obviously increased come of the animal experiment: 40 days after tumor necrosis factor-tumorinfiltrating lymphocytes infusion, cancer size in local tumor necrosis factortumor-infiltrating lymphocytes infusion group was found smaller than that in local infusion control group [(307±42) and (2 048±278) mm3, P ＜ 0.01],and it was also smaller in venous tumor necrosis factor-tumor-infiltrating lymphocytes infusion group than that in venous control group [(954±195)and (1 989±305) mm3 , P ＜ 0.05].CONCLUSION: Tumor necrosis factor-α gene transduced tumor-infiltrating lymphocytes could effectively express tumor necrosis factor, exerting higher and in vivo anti-tumor effects than tumor-infiltrating lymphocytes in cancer-loaded nude mice. No obvious inhibitory effects on the growth of subcutaneous solid carcinoma could be observed in nude mice after venous infusion of human brain glioblastoma tumor-infiltrating lymphocytes, but the inhibitory effects became obvious due to venous infusion of tumor necrosis factor-tumor-infiltrating lymphocytes and significant due to local tumor necrosis factor-tumor-infiltrating lymphocytes infusion, indicating that local infusion is the preferable way in the treatment of glioblastoma by immuno-gene therapy.