Allogenetic Stem Cell Transplantation And NK Cell Combined Bone Marrow Transplantation For The Treatment Of Prostate Cancer In Murine Model

Partâ… Non-myeloablative Allogenetic Bone Marrow Transplantation for the Treatment of Prostate Cancer in Murine ModelObjective The objectives of this part are to explore the graft versus tumor (GVT) effects and the severity of graft versus host disease(GVHD) post allogenetic bone marrow transplantation in murine prostate cancer model.Methods Following nonmyeloablative regimens consisted of fludarabine(30mg/m²,-d^1-5) and cyclophosphamide(300mg/m²,-d^1-3),C57BL mice bearing RM-1(2×10⁶ each) tumors underwent an allogeneic bone marrow cells(5×10⁶ each) and spleen cells(1×10⁷ each) transplantation from BALB/c donors through the tail veins.The tumor volumes and the survival time of the recipients were recorded and compared with the control group and the severity of GVHD was graded by pathology morphological standard.The bone marrow cells from the dying recipients were cultured for the evidence of bone metastasis.Results All C57BL mice were observed formation of transplanted prostate cancer.The nonmyeloablative regimens did not inhibit the growth of the tumors (P＞0.05).Allogenetic bone marrow transplantation inhibited the growth of the tumors and prolonged the recipients' survival time significantly(P=0.028 and 0.00001,respectively).The recipients' GVHD were gradingⅢ～Ⅳ,and the evidences of bone metastasis were found in the dying recipients.Conclusions Under these given nonmyeloablative regimens,GVHD were well endured by the recipients.Allogenetic bone marrow transplantation inhibited the growth of the tumors and prolonged the recipients' survival time significantly.The recipients' significantly prolonged survival time would be the main reason for the prostate cancer bone metastasis.Partâ…¡Non-myeloablative Allogenetic Bone Marrow Derived Mesenchymal Stem Cell Transplantation for the Treatment of Prostate Cancer in Murine ModelObjective The objectives of this part are to explore the graft versus tumor (GVT)effects and the severity of graft versus host disease(GVHD) post allogenetic bone marrow derived mesenchymal stem cell(MSC) transplantation in murine prostate cancer model.Methods Bone marrow cells were cultured and purified by adherent method to the fourth generation.The MSCs were then identified by flow cytometry for CD44 positive cells and oil red stain for the differentiated fat cells.Following nonmyeloablative regimens consisted of fludarabine(30mg/m²,-d^1-5) and cyclophosphamide(300mg/m²,-d^1-3),C57BL mice bearing RM-1(2×10⁶ each) tumors underwent an allogeneic bone marrow MSCs(2×10⁶ each) transplantation and bone marrow cells(5×10⁶ each) and spleen cells(1×10⁷ each) transplantation from BALB/c donors through the tail veins.The tumor volumes,survival time and the grade of GVHD of the recipients were recorded, and then compared with the allogenetic bone marrow transplantation group.Results The percentage of CD44 positive cells in the fourth generation was 84.29%,and the oil red stain proved the existence of differentiated fat cells. Comparing with the control group,allogenetic bone marrow derived MSC transplantation inhibited the growth of the tumors and prolonged the recipients' survival time significantly(P=0.047 and 0.00001,respectively).Comparing with the allogenetic bone marrow transplantation group,allogenetic bone marrow derived MSC transplantation shared the same inhibiting effect of the tumors and prolonging effect of the recipients' survival time(P=0.46 and 0.96, respectively).The MSC transplantation did not avoid the occurrence of GVHD, and the recipients' GVHD were gradingⅡ～Ⅲ,better than these of the allogenetic bone marrow transplantation group's(P=0.0017).Conclusions The method of MSC culture and identity was well founded. Allogenetic bone marrow derived MSC transplantation inhibited the growth of the tumors and prolonged the recipients' survival time significantly.Compared with allogenetic bone marrow transplantation group,the MSC transplantation shared same graft versus tumor effects,but decreased the severity of GVHD.Partâ…¢Alloreactive NK Cells Transplantation Enhanced GVT Effects and Decreased GVHD in Murine Prostate Cancer Model.Objective The objectives of this part are to explore the enhancement of the graft versus tumor(GVT) effects and reduction of the graft versus host disease (GVHD) post allogenetic bone marrow and alloreactive natural killer(NK) cell transplantation in murine prostate cancer model.Methods The alloreacitve NK cells were isolated from the spleen of CB6F1^H-2d/b(the F1 generation of C57BL male mice and BALB/c female mice) mice,purified by negative magnetic cell sorting(MACS),and then Ly49A positive cells were identified and collected by flow cytometry.Following nonmyeloablative regimens consisted of fludarabine(30mg/m²,-d^1-5) and cyclophosphamide(300mg/m²,-d^1-3),C57BL mice bearing RM-1(2×10⁶ each) tumors underwent an allogeneic bone marrow cells(5×10⁶ each) and alloreacitve NK cells(1×10⁶ each) transplantation 48 hours later from CB6F1^H-2d/b donors through the tail veins.The tumor volumes and the survival time of the recipients were recorded,the severity of GVHD was graded by pathology morphological standard,and the bone marrow cells from the dying recipients were cultured for the evidence of bone metastasis,and then compared with that of the allogenetic bone marrow transplantation group.Results The percentage of Ly49A positive cells in NK cells isolated from the CB6F1^H-2d/b mice was 44.98%.Compared with allogenetic bone marrow transplantation group,the alloreacitve NK cells transplantation inhibited the growth of the tumors and prolonged the recipients' survival time more significantly(P＜0.05 and 0.001,respectively);decreased the severity of the recipients' GVHD,which gradingⅠ～Ⅲ,better than these of the allogenetic bone marrow transplantation group's(P=0.0003);and the bone marrow culture of the dying recipients demonstrated evidences of bone metastasis inhibition.Conclusions Compared with allogenetic bone marrow transplantation, alloreactive NK Cells transplantation inhibited the growth of the tumors, prolonged the recipients' survival time and decreased the severity of GVHD more significantly.There are evidences that alloreactive NK cells transplantation inhibited the prostate cancer bone metastasis.Therefore,the alloreactive NK cells transplantation enhanced GVT effects and decreased GVHD in murine prostate cancer model.