Study On The Inhibition Of Colorectal Cancer Liver Metastasis By Injected Allogeneic Bone Marrow Cells And Lymphocyte

Objective:Using allogeneic hematopoietic stem cell transplantation(Allo-HSCT)for the prevention and treatment of relatively early stage of the colorectal cancer liver metastasis murine model.The model murine were injected with cyclophosphamide after transplanted with semi-allogeneic bone marrow and lymphocytes,then were transplanted with semi-allogeneic lymphocytes.Firstly the relationship between infusion time point of the allogeneic bone marrow cells combined lymphocyte and anti-colon cancer liver metastasis effect was established,then to investigate the inhibiting effect and condition of the chimera level and GVHD of the colorectal cancer liver metastasis murine model received allogeneic hematopoietic stem celltransplantation(Allo-HSCT),and to study its tumor suppressor mechanism,for providing a new method of immunotherapy for colorectal cancer liver metastases prevention.Methods:The bone marrow cells and lymphocytes of C57BL/6(H-2Kb)mice were used as donor,while(BALB/exC57BI/6)FI mice(CB6FI,H-2Kb/d)used as receptor.The CB6 FI were injected with CT26 cells in spleen to build the colorectal cancer liver metastasis murine model,randomly divided into four groups according to the different time of the caudal vein infusion with bone marrow cells and splenocytes.The clinical manifestation of GVHD,tumor liver metastasis and survival time of mice in each group were observed.(BALB/exC57BI/6)FI mice(CB6FI,H-2Kb/d),with CT26 cells injection in spleen were randomly divided into four groups,two of that were injected with bone marrow and lymphocytes from C57BL/6 mice CB6F1 mice from caudal vein at the best time according to the prior experiment.The survival condition,inhibition rate of liver metastasis and the GVDH after transplantation were observed.The chimeras were analysed by Flow Cytometer;The concentration levels of IL-2?INF-??IL-4?TGF-? were measured by ELISA.Results:1.The inhibition rate of the tumors and the survival period in the group which was injected into the caudal vein with bone marrow cells and splenocytes at 3 days after model built were significantly increased than the groups that was injected at 7days,14 days.But there was no significant difference in the GVHD score.The inhibition rate of liver metastasis of the group injected with cyclophosphamide after bone marrow and lymphocytes transplatation from C57BL/6 mice,followed by lymphocytes from C57BL/6 mice transplatation,was significantly higher than that in other groups,with the inhibition rate of the tumors and the survival period was significantly increased(P<0.05).Since 7 days after transplantation,the chimeric rate of the donor mouse cells gradually increased,until 28 days after transplantation,the receptor cells were almost replaced by the donor cells,the donor chimerism is more than 99%.At the same time,The GVT of the experimental mice was significantly enhanced with no serious GVDH symptoms.The concentration levels of IL-2?INF-? were higher than other groups,the concentration levels of IL-4 showed no obvious differences,the concentration levels of TGF-?were lower than other groups.Conclusion:Mice which were injected into the caudal vein with bone marrow cells and splenocytes at 3 days after model built can get a high tumor suppressor rate with no serious GVDH symptoms,and can live longer.The donor chimerism of the mice which transplanting bone marrow and lymphocytes from C57BL/6 mice after chemotherapy were significantly increased until 28 days after transplantation,with obvious GVT effect,bearing colorectal cancer liver metastasis.The change of the concentration levels of IL-2?INF-??IL-4?TGF-? were directly related to tumor growth inhibition.