Bone Marrow Mesenchymal Stem Cells Invovled In Prostate Carcinogenesis And Cancer Progress

ObjectiveTo investigate the optimal 137Cs radiation dose of Bone-marrow transplantation at rate of 0.708Gy/min,and establish the GFP Bone-rmarrow transplanted mice model.To observe the role of bone marrow-derived cells(BMDCs)in transplanted mice,and analysis the effect of bone marrow mesenchymal stem cells(BMSCs)to prostate cancer in vivo,discuss the mechanism of BMSCs promoting the development of prostate cancer.MethodC57BL/6 mice were irradiated with 7.5Gy,8.5Gy,9.5Gy,10.5Gy from 13'Cs respectively,then transplanted with new Bone-marrow cells within 4 hours,the mouse weight,survival rate and WBC in peripheral blood were obtioned.GFP Bone-marrow transplanted mice were established,GFP-positive cell rate of peripheral blood were detected in 4 weeks after transplantation,and rate of Bone-marrow cell in 28 weeks.TRAMP mice were verified by q-PCR and electrophoresis,the prostate carcinogenesis was acquired whit HE staining.rvMNU were injected in prostate of C57BL/6 mice every 3 weeks combining with testosterone propionate subcutaneous and intraperitoneal injection,lasting 6 monthes,the occurrence and progression of prostate cancer were evaluated with means of HE and immunohistochemical staining.GFP cells of transplanted Pca mouse model were detected by way of Small animals living imaging system,GFP immunohistochemical staining,immunofluorescence staining of Pca frozen sections.The cell which promotes the development of Pca in BMDCs was found by Flow cytometry techniques,and the relationship of BMSCs and RM-1 was obtioned through the formation of subcutaneously transplanted tumor.ResultsIrradiated mice exhibited acute radiation injury within lweeks,the peripheral blood leukocytes kept decreasing early,mice irradiated with 7.5Gy,8.5Gy begin to increase rapidly 7 days later,and were recovered completely 5 weeks after marrow transplantation.GFP-positive rate of peripheral non-erythroid cells was 75.2%-95.8%(mean:85.3%± 7.2%)4 weeks after marrow transplantation,and GFP-positive rate of bone marrow cells was 90.6%-99.9%(mean:95.2%± 2.5%)28 weeks after marrow transplantation.The tumor incidence of TRAMP was 55%(11/20)in 36 weeks,and tumor metastasis incidence was 20%(4/20).The tumor incidence of MNU-induced mice was 78%(39/50),PIN included,and no metastasis tumor was found,prostatic hyperplasia,intraepithelial neoplasia,primary and invasive carcinoma could be seen in HE staining of Paraffin sections.GFP can be expressed in a protion of Pea cancer cells,some Pea cancer cells had SV40-Tag/GFP(CK8/GFP)double-expression,with the methods of Small animal in vivo imaging system and IHC/IF.Flow cytometry of Pea single cell analysis showed the cells with GFP(+)/Sca-1(+)/CD45(-)increased significantly compared with the control group.The growth curve of RM-1+BMSCs group xenograft was significantly higher than RM-1 group alone,and CK8(+)/GFP(+)double positive cells appeared in RM-1+BMSCs group.The cytokines of orthotopic tumor and xenografts had differential expression of IL-6,IL-8,SDF-1,VEGF signifcantly.Conclusion8.5Gy is the optimal irradiation dose for bone marrow transpantation at rate of 0.708Gy/min in C57BL/6 mouse,GFP transplanted mice model is a appropriate tool for studying the BMDCs.C57BL/6 mouse under MNU injection in situ combining with testosterone propionate can progress into Pea,and it is similar to human progression.BMDCs promted Pea carcinogenesis and progression,and BMSCs played important roles in this process.Beside,BMSCs can help the xenograft growth and proliferation of Pea cell lines RM-1.Cytokines in microenvironment exerted some efforts in the process of carcinogenesis between the interaction of BMSCs and protate cells.