ABSTRACT
<p><b>OBJECTIVE</b>During the process of tissue remodeling in human tumor transplantation models, the roles of the inoculated tumor cells and host tissue in tumor progression is still largely unknown. The aim of this study was to investigate the relationships and interactions between these two sides using GFP-RFP double fluorescence tracing technique.</p><p><b>METHODS</b>Red fluorescence protein (RFP) gene was stably transfected into glioma stem cell line SU3, then SU3-RFP cells were transplanted into the brain of athymic nude mice with green fluorescence protein (GFP) expression. After the intracerebral tumors were formed, the relationship and interaction between GFP cells and RFP cells were analyzed. Highly proliferative GFP cells were screened out, and monocloned with micro-pipetting. DNA content assay, chromosome banding and carcinogenicity test of the GFP cells were performed to observe the GFP cells' cancerous phenotype in nude mice.</p><p><b>RESULTS</b>In the transplantable tumor tissue, besides a great quantity of RFP cells, there were still a proportion of GFP cells and GFP/RFP fusion cells. The proportion of RFP cells, GFP cells and GFP/RFP cells were (88.99 ± 1.46)%, (5.59 ± 1.00)%, and (4.11 ± 1.020)%, respectively. Two monoclonal host GFP cells (H1 and H9) were cloned, which demonstrated the properties of immortality, loss of contact inhibition, and ultra-tetraploid when cultured in vitro. Both H1 and H9 cells expressed CNP, a specific marker of oligodendrocytes. The GFP cells also demonstrated 100% tumorigenic rate and high invasive properties in vivo.</p><p><b>CONCLUSIONS</b>In this glioma transplantation model, the transplanted tumor tissues contained not only transplanted glioma stem cells but also cancerous host GFP cells. Our findings offer important clues to further research on the relationships among different members in the tumor microenvironment.</p>
Subject(s)
Animals , Humans , Mice , 2',3'-Cyclic Nucleotide 3'-Phosphodiesterase , Metabolism , Brain , Cell Biology , Metabolism , Cell Communication , Cell Line, Tumor , Cell Transformation, Neoplastic , Glioma , Metabolism , Pathology , Green Fluorescent Proteins , Metabolism , Intermediate Filament Proteins , Metabolism , Luminescent Proteins , Genetics , Metabolism , Mice, Inbred C57BL , Mice, Nude , Neoplasm Transplantation , Neoplastic Stem Cells , Cell Biology , Metabolism , Nerve Tissue Proteins , Metabolism , Nestin , Neuroglia , Cell Biology , Metabolism , Transfection , Tumor MicroenvironmentABSTRACT
<p><b>OBJECTIVE</b>To investigate the autoimmune injuries of diabetic macrovascular disease (aorta) and the protective effects of immunosuppressive agent (cyclosporine A, CsA) on aortic injuries in streptozotocin (STZ)-induced diabetic rats.</p><p><b>METHODS</b>STZ-induced diabetic rats were assigned randomly to 6 groups which received low (BML or AML, 1 mgxkg(-1)xd(-1)), middle (BMM or AMM, 4 mgxkg(-1)xd(-1)) or high (BMH or AMH, 8 mgxkg(-1)xd(-1)) dose of CsA from 1 week before or after STZ for 8 weeks. Diabetic rats without any treatment, insulin-treated diabetic rats and normal rats were also monitored simultaneously and served as control groups. The pathologic abnormalities of the aorta were verified by HE, Masson staining and electronmicroscopy. The depositions of immunoglobulins (IgG, IgM and IgA) were determined by immunohistochemistry and immunofluorescence methods.</p><p><b>RESULTS</b>At the end of study, lymphocytes infiltration and collagen content (26 582 +/- 6901) were significantly higher in diabetic aorta than those in non-diabetic aorta (Collagen: 7482 +/- 3491, P < 0.01). The deposited IgG and IgA were also significantly increased in diabetic aorta compared with non-diabetic aorta (IgG: 11 789 +/- 2491 vs. 2518 +/- 1066, P < 0.01; IgA: 17 430 +/- 3159 vs. 1135 +/- 758, P < 0.01). These changes were not affected by insulin while CsA intervention significantly reduced aortic collagen content (BMH: 13 518 +/- 5440, P < 0.01 vs. STZ) and immunoglobulin deposition (BMH: IgG: 7584 +/- 4462; IgA: 6176 +/- 1900, all P < 0.01 vs. STZ). These immunoglobulin deposition changes were confirmed by results of immunofluorescence. Aortic collagen accumulation was positively correlated to aortic immunoglobulin deposition (IgG, r = 0.556, P < 0.01; IgA, r = 0.661, P < 0.01).</p><p><b>CONCLUSIONS</b>Our data suggest that the autoimmune injuries might be a promoting factor in the pathogenesis of the diabetic macrovascular disease which could lead to the development of macrovascular disease. Immunosuppressive agent, such as CsA, could inhibit the abnormal deposition of immunoglobulins and therefore, delay the development of diabetic macrovascular disease in this model.</p>
Subject(s)
Animals , Rats , Aorta , Allergy and Immunology , Pathology , Aortic Diseases , Cyclosporine , Pharmacology , Diabetes Mellitus, Experimental , Allergy and Immunology , Pathology , Endothelium, Vascular , Pathology , Immunosuppressive Agents , Pharmacology , Rats, Sprague-DawleyABSTRACT
The depositions of immunoglobulins on the retina of diabetic rat were studied with immunohistochemistry,immunofluorecence and computer analysing system.In comparison with control group, depositions of IgG,IgA and IgM on the retina of diabetic rat induced by streptozotocin were significantly increased (all P＜0.05 ).So the immunoglobulin depositions may contribute to the pathogenesis of diabetic retinopathy.