Treatment efficacy of DC-CIK with different loading combined with 5-FU on nude mice bearing colon cancer HT-29 cell transplanted tumor / 中国肿瘤生物治疗杂志

@# Objective: To investigate the therapeutic effect of dendritic cell-induced killer cells (DC-CIK) combined with 5-fluorouracil (5-FU) and loaded with CD133+ HT-29 cell lysate or RNA on mice bearing colon cancer HT-29 cell transplanted tumor, and to explore the underlying mechanism. Methods: Colon cancer xenograft model was established in BALB /c nude mice by using human colon cancer HT-29 cells at logarithmic growth phase; Antigen-free DC-CIK, 5-FU+DC-CIK, R+DC-CIK (loaded with total RNA of CD133+ cells), L+DC-CIK (loaded with CD133+ cell lysate), 5-FU and normal saline were respectively injected into transplanted mice, and the treatment efficacies on the growth of transplanted tumor in each group after three treatment cycles were observed, and the tumor growth curve was drawn. The nude mice were sacrificed by cervical dislocation and the tumor volume and body weight were measured. qPCR was used to detect the expression of AKT mRNA in transplanted tumor tissue, and WB was used to detect the expression of phosphorylated AKT protein. Results: After treatment, the body mass of nude mice in R+DC-CIK group, L+DC-CIK group and 5-FU+DC-CIK group increased steadily, while the body mass of nude mice in DC-CIK group and 5-FU group decreased gradually; the tumor growth speed of nude mice in R+DC-CIK group, 5-FU+DC-CIK group and L+DC-CIK group was significantly slower than that of the control group (P<0.05). Compared with 5-FU and DC-CIK alone, the combined treatment with loaded lysate/RNA had more sig nificant effect on mRNA and protein expressions of AKT(P<0.05). Conclusion: The effect of DC-CIKwithdifferentloadingoritscombinationwith5-FUisbetterthanthatofchemotherapy alone. One of the mechanisms is related to the down-regulation ofAKT level.

Clinical efficacy of DC-CIK immunotherapy in treatment of advanced pancreatic adenocarcinoma / 中国肿瘤生物治疗杂志

@#［ ［Abstract］ ］Objective: To analyze and compare the clinical efficacy and safety of dendritic cell cytokine-induced killer cells (DCCIK) combined with palliative therapy or chemotherapy in the treatment of advanced pancreatic carcinoma. Methods: A retrospective study was carried on 50 patients with advanced pancreatic carcinoma who were hospitalized in department of oncology of Shanxi Dayi Hospital during September 2012 to February 2016. The patients were divided into four groups according to the therapy they received (palliative treatment group, palliative+DC-CIK treatment group, chemotherapy group and chemotherapy+DC-CIK treatment group); the immunological function, quality of life and survival time of patients were analyzed; and the efficacy and safety of DC-CIK cell therapy was also evaluated. Results: The percentages of CD8+ T cells and NKT cells in DC-CIK combined therapy groups were significantly improved compared with that of pre-treatment, and the percentages of CD3+, CD8+, NK, NKT cells were increased compared with control groups (P<0.05). The quality of life of patients was significantly improved (P<0.05), while median PFS and median OS were improved but without statistical significance (P>0.05). Conclusion: Compared with palliative therapy and chemotherapy alone, combined DC-CIK immunotherapy can effectively improve the cellular immunity function and quality of life in patients with advanced pancreatic cancer. However, there was no significant extension in overall survival.

Killing effect of Huaier combined with DC-CIK on nude mice bearing colon cancer HT29 stem cells / 中国中药杂志

To compare the therapeutic effects of different treatment methods on the nude mice bearing colon cancer HT29 cells. BalB/C nude mice colon cancer stem cell models were established and randomly divided into the following four groups, with 8 nude mice in each group: blank control group, DC-CIK group, Huaier group, and Huaier combined with DC-CIK group (combined treatment group). The mice in DC-CIK group and combined treatment group received 1×10⁶ DC-CIK cells treatment by tail vein injectionafter the tumor stem cells were inoculated for 4 days,2 times a week for three weeks. The mice in Huaier group and combined treatment group received intragastric administration at the dose of 20 g/60 kg body weight, 0.2 mL/time, once a day for a total of three weeks. The mice in control group received equal volume of normal saline. Tumor size and body weight of nude mice were measured every 2 days during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to measure the tumor weight and the tumor inhibition rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in the signal pathway. After the end of the treatment, the quality of the tumor in the Huaier group, DC-CIK group and combined treatment group was significantly lower than that in the control group; the quality in combined treatment group was significantly lower than that in Huaier group and DC-CIK group.Among them, the tumor inhibition rate reached 46.77% in the combined treatment group. In respect of changes in expression levels of key genes in the signaling pathway, the mRNA expression levels of key genes PI3KR1 and Akt in PI3K/Akt pathway, key genes Wnt1 and CTTNB1 in Wnt/-catenin pathway, and key genes Notch1, Notch2, Notch3 in Notch pathway in the combined treatment group were lower than those in DC-CIK group and Huaier group. The Huaier combined with DC-CIK group showed best therapeutic effect among different treatment methods for HT29 stemcell colon tumors in nude mice, providing a new idea for clinical treatment of colon cancer.

In vivo antitumor effect of Chinese yam extract combined with DC-CIK on nude mice bearing colon cancer HT29 stem cells / 中草药

Objective: To compare the therapeutic effects of different treatments on the nude mice bearing colon cancer HT29 cells. Methods: BalB/C nude mice colon cancer stem cell model was established and the mice were randomly divided into following four groups: blank control group, DC-CIK group, yam group, and yam combined with DC-CIK group (combined treatment group), each group of 10 nude mice. In DC-CIK group and combined treatment group, after tumor stem cells were inoculated in nude mice for 4 d, 1 × 106 DC-CIK cells were used through the tail vein injection for treatment, twice a week for three weeks. The mice in Chinese yam group and combined treatment group were ig administered with 125 mg/kg Chinese yam extract, once daily for three weeks, and the mice in control group were treated with equal volume of saline instead. Tumor size and body weight of nude mice were measured every 2 d during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to take out the tumor weight and the tumor inhibitory rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in signal pathway. Results: After treatment, the quality of tumor in yam group, DC-CIK group and combined treatment group was significantly lower than that in control group. Among them, the inhibitory rate was 51.26% in combined treatment group. In the changes of expression levels of key genes in signaling pathway, the key genes of PI3KR1 in PI3K/Akt pathway, the key gene of Wnt1 in Wnt/beta-catenin pathway, and the mRNA expression of key gene Notch1 in Notch pathway in combined treatment group were decreased compared with DC-CIK group and yam group. Compared with the DC-CIK group, the mRNA expression changes were not statistically significant. Conclusion: The effect of Chinese yam combined with DC-CIK group is the best on the treatment of HT29 cell stem cell tumor of colon cancer in nude mice. It provides a new idea for clinical treatment of colon cancer.

Therapeutic effect of DC-CIK cells combined with S-1 in the treatment of non-small-cell lung cancer / 国际生物医学工程杂志

Objective To observe the clinical effect of DC-CIK cells combined with S-1 in the treatment of non-small-cell lung cancer.Methods 76 patients with non-small-cell lung cancer were randomly divided into the observation group and the control group on average.The control group was treated with S-1,and the observation group was treated with DC-CIK cells combined with S-1.For the observation group,the peripheral venous blood was collected before the treatment for DC cells and CIK cells cultivation.The expression of CD4+/CD8+,CD4+ and NK cells in the peripheral blood of the two groups was detected by flow cytometry before the treatment and after one week of the treatments.Besides,the number of white blood cells and platelet count were also measured and the symptoms of non-small-cell lung cancer were observed.Results The percentage of CD4+/CD8+,CD4+ and NK cells in the peripheral blood of the observation group after the treatment was (1.65±1.03),(34.56±8.90) and (18.68±7.98),respectively,which was significantly higher than (1.32±0.70),(29.07±7.15) and (15.28±8.23) in the control group (all P＜0.05).There was no significant difference in the percentage of CD8+ cells between the observation group (25.56± 8.90) and the control group (26.64±6.77) (P＞0.05).The vomiting,leukopenia and thrombocytopenia in the observation group were less than those in the control group (all P＜0.05).The one-year survival rate was 52.63％ in the observation group which was significant higher than 42.11％ in the control group (P＜0.05).Conclusions DC-CIK cells combined with S-1 is a safe and effective treatment for non-small-cell lung cancer,which can effectively improve the clinical efficacy and prolong the survival time of patients.

The relationship between DC-CIK and colorectal cancer prognoses and its influen-cing factors / 药学实践杂志

Objective To discuss dendritic cell-cytokine-induced killer (DC-CIK ) cell therapy effects and clinical out-comes in patients with colorectal cancer in order to have better clinical treatment .Methods A retrospective analysis of the data of 66 patients with colorectal cancer from the Biological Therapy Department of the Eastern Hepatobiliary Surgery Hospital was performed from January 2012 to January 2014 ,and then was followed up .Taking gender ,age ,degree of pathological differen-tiation ,TNM staging ,surgical methods ,and targeted therapy as the research basis ,by the Kaplan-Meier single factor and Cox multiple factors analysis we mainly discuss the DC-CIK cell treatment′s effect on the prognoses of patients .Results Kaplan-Meier single factor analysis results indicate :to a certain extent ,DC-CIK cell therapy can improve the prognoses of patients ;Cox multi-factor analysis results indicate whether accepting DC-CIK cell therapy is an independent factor influencing the prog-noses of patients .Conclusion DC-CIK cells therapy can improve the prognoses of patients with colorectal cancer .

Clinical controlled trial of the advanced gastric patients with ascites by intra-abdominal infusion of DC-CIK or rmhTNF / 实用肿瘤学杂志

Objective This study is to explore the differences in the curative effect of intra -abdominal infusion between Recombinant mutant human tumor necrosis factor and Dendritic cell -Cytokine induced killer . Methods We selected 48 advanced gastric cancer patients with ascites .Those patients were randomized into two groups:DC-CIK group and rmhTNF group .After one month treatment ,we observed the adverse events both two groups,and evaluated the clinical beneficial responses ,the responsive rate ,the tumor indicators′level,the immune indexes and the time of tumor progression .Results In the DC-CIK group,the RR and the CBR were 83.3%and 66.67%,respectively;while in the rmhTNF group,they were 58.33%and 75%,respectively.The difference between two groups was statistically significant (P0.05).The ratios of CD3 +、CD4 +、NK cells distinctly increased after treatment (P0.05).One year follow-up of the time to progression(TTP)was 7.1 months in DC-CIK group,and 5.8 months in TNF group,which was statistically significant (P=0.02).Conclusion Both rmhTNF and DC-CIK can improve the efficacy for the patients with malignant ascites caused by gastric cancer .However ,DC-CIK immunotherapy shows better active effect on improvement of specific immune function .

Transarterial chemoembolization combined with autologous DC-CIK cells for the treatment of hepatocellular carcinoma of BCLC C-stage：a randomized controlled study / 介入放射学杂志

Objective To investigate the efficacy of transarterial chemoembolization (TACE) combined with autologous DC-CIK cells in treating hepatocellular carcinoma(HCC) of BCLC C-stage. Methods A total of 60 cases with HCC in BCLC C-stage were randomly and equally divided into the study group (n=30) and the control group (n=30). TACE combined with autologous DC-CIK cells was employed in the patients of the study group, while only TACE was adopted in the patients of the control group. The immune function, six-month and one-year survival rates were determined, and the results were compared between the two groups. Results In the study group, the blood T lymphocyte subsets of CD3+CD8+ were significantly increased, while CD3+CD4+ were obviously decreased. When compared with the pretreatment levels, the differences were statistically significant (P<0.05). The six-month survival rate of the study group and the control group was 67.9% and 48.1% respectively (P<0.05), and the one-year survival rate of the study group and the control group was 53.6%and 29.6%respectively (P<0.05). Conclusion For the treatment of HCC in BCLC C-stage, the therapeutic effect of TACE combined with autologous DC-CIK cells is much better than that of pure TACE. Therefore, this therapy is an effective treatment for HCC in BCLC C-stage.

Effect of DC-CIK cells combined with oridonin on cytotoxicity against RPMI 8226 cells / 实用医学杂志

Objective To investigate the changes in cytotoxicity of DC-CIK cells to human multiple myeloma RPMI 8226 cells before and after treatment with oridonin. Methods Normal human peripheral blood mononuclear cells were isolated and induced to obtain DC-CIK cells. Cytotoxicity of DC-CIK cells against RPMI 8226 cells which were treated by oridonin was analyzed by LDH releasing assay. The variation for expression of NKG2D ligands on RPMI 8226 cells were measured by flow cytometry. Results DC-CIK cells were successfully induced from the peripheral blood mononuclear cells. At the same effector to target ratio, oridonin obviously enhanced the cytotocixity of DC-CIK cells against RPMI 8226 cells (P<0.01). Flow cytometry showed the expression of NKG2D ligands ULBP1 of RPMI8226 cells was most significantly increased as the cells were treated by oridonin [(9.19 ± 1.85) vs. (15.47 ± 0.67), P<0.01]. Correlation analysis indicated that cytotocixity was positively correlated with changes in ULBP1. Conclusions Oridonin can improve the cytotoxicity of DC-CIK cells against RPMI 8226 cells, which may be related with the increased expressions of NKG2D ligands on the tumor cell surface.

Mechanism of lymphocyte function-associated antigen-1/intercellular adhesion molecule-1 mediated anti-neoplastic effects of cytokine-induced killer cells / 白血病·淋巴瘤

Objective To investigate the molecular mechanism underlying lymphocyte functionassociated antigen-1 (LFA-1) / intercellular adhesion molecule-1 (ICAM-1) mediated anti-neoplastic effects of cytokine induced killer (CIK) cells. Methods Lymphocytes isolated from peripheral blood of children leukemia were induced with interferon-gamma (IFN-y), anti-CD3 monoclonal antibody (CD3McAb) and interleukin-2 (IL-2) and co-cultured with dendrite cells (DC) to generate DC-CIK cells. When treated with LFA-1 monoclonal antibody, cytotoxicity of DC-CIK cells against leukemia cell lines was measured by the MTT assay, while RT-PCR and Western blotting were used to determine mRNA and protein expressions of GATA-3 and T-bet in DC-CIK cells, respectively. IL-12, IFN-γ and tumor necrosis factor-α (TNF-α) levels released by DC-CIK cells were quantified by ELISA. Results Induced DC-CIK cells were regular, round and transparent with variable cell volume and cellular aggregation. When treated with mouse anti-human LFA-1 monoclonal antibody, the cytotoxicity decreased mostly towards B95 cells under administration of 20 μg/ml LFA-1 monoclonal antibody in comparison with the control group(t =10.138, P ＜0.05). It led to a highest elevation of GATA-3 mRNA and protein levels (t =16.386, P ＜ 0.05; t =22.652, P ＜ 0.05) and a most decrease of T-bet mRNA and protein levels (t =17.728, P ＜0.05; t =17.452, P ＜0.05) under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group. The expression levels of IL-12,IFN-γ, and TNF-o in supernatant were the lowest under 20 μg/ml LFA-1 monoclonal antibody in B95 cells group in comparison with the control group (t =21.621, P ＜0.05; t =13.739, P ＜0.05; t =15.278, P ＜0.05).Conclusion GATA-3 and T-bet were implicated in the LFA-1/ICAM-1 mediated anti-neoplastic effects of DC-CIK cells via activation of the Th1 pathway, with high secretion of Th1 cytokines, such as IL-12, IFN-γ and TNF-α.