The Effects Of CIK Cells On Hypoxia Inducible Factor-1a(HIK-1a) And T Cell Subsets In Colon26Cancer Xenograft Mice

Objective: Cytokine induced killer cells (CIK) is a kind of immuneactive cells which is fast breeder, wide antineoplastic spectrum and highantitumor activity. They can promote the reconstruction of the patients’immune system and eliminate the residual lesions, which have certain curativeeffect for a variety of malignant tumors. Chemotherapy has bone marrowtoxicity, which is easy to cause the immunosuppression and adverse to theimmunotherapy. However, recent studies have found that there are not onlydirectly cytotoxic function, but also used as adjuvant in antitumor immuneresponse in many chemotherapy drugs. Immunotherapy has the potential ofenhance the sensitiveness of chemotherapy. It has the potential of mutualsynergy between chemotherapy and immunotherapy. The synergisticmechanism is complex between immunotherapy and chemotherapy, for whichchemotherapy and immune escape for more research. It has few researchabout tumor hypoxia microenvironment influence between immunotherapyand chemotherapy now. Los platinum (LBP) is the third generation ofplatinum antitumor drugs, quick effect, long duration, high concentration inthe tumor tissue and low in plasma. This study detect the the expression ofhypoxia inducible factor-1a (HIF-1a) and T cell subgroup in colon26tumor-burdened mice. Discussion the relationship between CIK andtumor-burdened hypoxia microenvironment、immunity, then explore the theinfluence of platinum and CIK joint application to both of tumor-burdenedhypoxia microenvironment and immunity. To provide theoretical basis for theclinical application of chemotherapy combined with immunotherapy.Methods:1Establish murine colon26carcinoma tumor models,With the processi-ng of CIK、LBP+CIK、LBP,and normal saline (NS) treatment as control group from the first eight days.Put the mice to death after treatment for1week,stripping tumors, weigh tumor weight respectively and take out the smallintestine, spleen tissue.2Detection HIF-1a mRNA of tumor tissue and the small intestine in miceby RT-PCR.3Detection the percentage of CD3+、CD4~+and CD4~+/CD8~+ratio in micespleen by FCM.4Changes of the surface of the small intestine in each group wereobserved by Scanning electron microscopy.5Analyze the data with SPSS16.0.Quantitative data were expressed asmean±standard deviation (χ±s), the two groups were compared using the t-test,P≤0.05for the difference was statistically significant.Results:1Effects of CIK and LBP on tumor volumes:Compared with NSgroup,tumor volume in CIK group are reduced, but not statistically significant(P>0.05);tumor volume in LBP group and LBP+CIK group are reduced,statistically significant(P<0.05);compared with CIK group,tumor volume shrinkagein LBP group are more obvious, statistically significant(P<0.05);comparedwith CIK group, tumor volume in LBP+CIK group are statistically significant(Ρ<0.05)(Table1).2Changes of HIF-1a gene expression and T lymphocytes in spleen ofmice: there were HIF-1a gene expression both in tumor tissue and smallintestine in Xenograft mice,and HIF-1a gene expression in tumor tissue wassignificantly higher than the small intestine (P<0.05),but its expression wasnot seen in the small intestine of healthy mice(Figure3,4,Table2);spleenCD4~+T-cell percentage and CD4~+/CD8~+T ratio of xenograft mouse wassignificantly lower than that in normal mice (P<0.05)(Figure5-9,Table3).3Effect on HIF-1a gene expression in tumor tissue by treatment of CIKand LBP: Compared with the NS group, the HIF-1a gene expression of micetumor tissue in CIK group decreased,but not statistically significant (Ρ>0.05);HIF-1a gene expression have reduced both in LBPgroup and LBP+CIK group, were statistically significant; compared with LBP group, LBP+CIK groupsignificantly decreased, there is a statistically significant (Ρ<0.05); comparedwith the CIK group, HIF-1a in LBP group significantly decreased, statisticallysignificant (Ρ<0.05)(Table2)4Influence of CIK and LBP on spleen T lymphocytes in xenograftmice:Compared with the NS group, CD4~+T-cell percentage is higher, whilethe CD8~+T-cell percentage is lower in CIK group, there is statisticallysignificant (P <0.05); compared with the NS group, CD4~+T cell percentage islower, and CD8~+T cell percentage is higher in LBP group, it has statisticallysignificant (P <0.05); compared with the NS group, CD4~+T-cell percentage ishigher, CD8~+T-cell percentage is lower in LBP+CIK group, there is nostatistically significant (Ρ>0.05). T lymphocytes in CIK group are higher thanLBP+CIK group and LBP group (P<0.05)(Fig.5,9, Table3).5Effects of CIK and LBP on small intestine mucosal in mice: Asscanning electron microscopy showed, compared with the NS group, smallintestinal in CIK mice mucosal thickness, the midgut gland depth, villusheight and surface area increases(Fig.10,11); the small intestinal mucosalthickness, the depth of the midgut gland, villus height and surface area toreduce in LBP group(Fig.12,13); pathological changes of small intestinalmucosal between LBP+CIK group and NS group has little change.(Fig.14,15).Conclusion:1Colon26transplanted tumor mice, whose immune function is decreased,can be detected HIF-1a both in tumor and small intestine tissue. Los platinumchemotherapy could further reduce immune function of transplanted tumormice.2It can improve the microenvironment of tumor hypoxia and promoteimmune reconstitution, and enhance the the CIK anti-tumor effect by the wayof combined with LBP.3CIK can improve the intestinal mucosal injury produced by tumor andchemotherapy.