The Preliminary Study On The Relationship Between Tramp Prostate Cancer And Bone Marrow-derived Stem Cells

Objective:TRAMP prostate cancer in mice bone marrow transplantation animal model is established,the preliminary discussion on relationship between prostate cancer and bone marrow-derived stem cells,in order to provides the basis of to further clearly the mechanism of prostate cancer and various epithelial cancer,and provide theoretical study basis for its treatment.Methods:Genotype identification of the TRAMP mice and immunohistochemical SV40T antigens.20 C57BL/6 n mice according to different accept CPL cs irradiation doses of gamma radiation is divided into four groups,and evaluate different dose of C57BL/6n mice,to formulate clear pulp exposure dose.6 weeks of green fluorescent protein(GFP)transgenic C57BL/6 j mice with strain C57BL/6 n and normal mice as a donor bone marrow cells,6 to 8 xweeks of age 15 only TRAMP transgenic mice as a receptor.The humerus and femur surgical separation of GFP mice,tore to muscle,cut to the epiphyseal end,rinse marrow cavity for bone marrow cells,red blood cells and backup on the ice.Receptors in mice receiving 137 cs lethal doses of gamma radiation exposure and radiation dose was 8.5 Gy,dose rate is 0.708 Gy/min,2 hours after irradiation per mice after injection of 2 x 105 bone marrow mononuclear cells.Accept 5 weeks after bone marrow transplantation,chimeric mice bone marrow rebuilding is confirmed by the flow cytometry instrument detection model is established successfully.Receptors in rat breeding SPF environment to 28 weeks,all 20 mice death,separation of bone marrow cells,further confirmed that bone marrow transplantation model was established.Separation of prostate tissue paraffin embedding,the line 5 microns section of HE staining,judge the TRAMP mice into tumors or not.And through the immunochemical staining to observe the GFP organization GFP positive cells forming the glands is proportion of prostate cancer.Results:TRAMP in mice by SV40 sequence PCR identification,are all TRAMP mice,put to death in mice after SV40T antigen immunohistochemical results are highly expressed T antigen.Dose acuity 8.5 Gy group and 7.5 Gy death numerical difference of white blood cells in mice significantly(P<0.01),acuity 8.5 Gy group all die of hematopoietic failure,and with the increase of irradiation dose,survival period shortened,while 7.5 Gy group survived for a long time;Of hematopoietic failure time and the maximum weight loss also have difference between each group respectively(P<0.01,P<0.05),along with the increase of irradiation dose,shorter time of hematopoietic failure,reduce the maximum weight gain.5 weeks after bone marrow transplantation,peripheral blood flow cytometry instrument detection showed that GFP positive cells from the bone marrow to the percentage of not less than 80%;28 weeks after bone marrow transplantation,peripheral blood flow cytometry Instrument test showed that all the TRAMP GFP positive cells from mouse bone marrow were greater than 80%,the percentage of GPF positive cells in the bone marrow can be accounted for about 90%above.10 only 28 weeks TRAMP prostate tumors in mice tumor rate was 100%(10/10).General observations is shown in figure 2,visible large prostate cancer.lung nodular metastases and nodular liver metastases.Incidence of pulmonary nodular lesions at 6/10,4/10 cases with nodular liver metastases.HE staining microscopic observations is shown in figure 3,and visible well-differentiated adenocarcinoma and poorly differentiated carcinoma.Prostate specimens on the TRAMP mice lines of GFP antibody immunohistochemical,GFP expression in prostate gland epithelium obviously positive.Conclusion:(1)TRAMP to spontaneous form of prostate cancer in mice,and canform the liver and lung metastases.(2)the137 cs illuminate,gamna radiation dose rate was 0.708 Gy/min,the dose of 8.5 Gy for min qing pulp exposure dose,can successfully induce bone marrow suppression,and bone marrow transplantation model was implemented.(3)the bone marrow-derived stem cells in the occurrence,development and metastasis of prostate cancer play a role in the process.