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Objective To investigate the proteasome inhibitor boron bortezomib against multiple myeloma (multiple myeloma, MM) patients with bone marrow mesenchymal stem cells (bone marrow mesenchymal stem cells, BMMSC) inhibition of growth and proliferation, and its ability to chemotactic and chemoattractant the related genes monocyte chemoattractant protein -1 (monocyte chemotactic protein-1, MCP-1), stromal cell-derived factor -1 (stromal cell derived factor-1, SDF-1), vascular endothelial growth factor (vascular endothelial growth factor , VEGF), hepatocyte growth factor (hepatocyte growth factor, HGF), insulin-like growth factor-1 receptor (insulin-like growth factor-1 receptor, IGF-1R), platelet-derived growth factor receptor α (platelet-derived growth factor receptor alpha, PDGFRα) mRNA expression levels. To reveal MM patients BMMSC, biology abnormalities and the mechanisms of the bone marrow microenvironment bortezomib. Collected eight cases the onset of bone marrow samples of MM patients and 11 normal controls by density gradient centrifugation and cultured in vitro BMMSC to take P2-4 generation BMMSC used for subsequent experiments. The experiment was divided into four groups, the namely MM group BMMSC, normal control group BMMSC, the MM group BMMSC bortezomib treatment and control group BMMSC. Use different concentrations of bortezomib on the growth inhibition rate BMMSC MTT assay, Annexin V / PI staining flow cytometry bortezomib of BMMSC apoptosis, using real-time quantitative PCR method BMMSC MCP-1, SDF -1, VEGF, HGF, IGF-1R, PDGFRαmRNA expression levels, while the establishment of MCP-1, SDF-1, VEGF, HGF, IGF-1R, PDGFRα and internal reference glyceraldehyde -3 - phosphate dehydrogenase (glyceraldehyde-3- phosphate dehydrogenase, GAPDH) of the standard curve, the amount of the relative expression of the target gene expressed in the copy number of the objective gene copy number / GAPDH. Transwell model to detect group BMMSC migration. The results of a study found on the MM group BMMSC morphology is no different, are spindle-shaped fibrous cells with the normal control group BMMSC. MTT method and Annexin V / PI staining and flow cytometry bortezomib can inhibit the growth and proliferation of BMMSC the ability and promote the apoptosis and growth inhibition rate in a time and dose-dependent increase in bortezomib for 48 hours on the of normal controls BMMSC the growth inhibition was stronger than the MM group BMMSC the (P lt; 0.01). By real-time PCR method to detect group BMMSC of MCP-1, SDF-1, VEGF, HGF, IGF-1R, PDGFRαmRNA expression level results show that the MM group BMMSC to abnormal cytokine expression in the MM group BMMSC SDF-1 mRNA expression compared with normal control group (P lt; 0.05), VEGF, HGF mRNA expression compared with normal control group (P lt; 0.05). Bortezomib may reduce the MM group BMMSC and normal control group BMMSC of MCP-1, SDF-1, VEGF, HGF, IGF-1R, PDGFRαmRNA relative expression levels were statistically significant (P lt; 0.05), bortezomib Zuo meters the two BMMSC above inhibition of gene expression was no significant difference (P gt; 0.05). Transwell migration model checking results, the the MM group BMMSC and the normal control group BMMSC chemotactic ability was no significant difference (P gt; 0.05). The boron on behalf of the bortezomib role after MM group BMMSC and normal control group BMMSC chemoattractant significantly reduced (P lt; 0.05). With or without bortezomib treatment, SDF-1 can increase MM group BMMSC and normal control group BMMSC migration (P lt; 0.05), especially for MM group BMMSC more significantly (P lt; 0.05). The Conclusion MM group BMMSC the compared with normal control group BMMSC, are spindle-shaped fibrous cells morphology, no significant difference in the ability of the two chemokines, chemokine SDF-1 under the action of the chemoattractant ability can be improved, but the MM group BMMSC the 's of SDF-1, VEGF, HGF mRNA expression level exception, suggesting that patients with myeloma in the bone marrow microenvironment BMMSC appear abnormal chemotactic factor expression, perhaps with myeloma cell homing and colonization, growth, resistance-related. Bortezomib inhibit BMMSC growth and proliferation and chemotactic ability, while reducing MCP-1, SDF-1, the expression levels of VEGF, HGF, IGF-1R, PDGFRαmRNA, suggesting bortezomib addition to the direct effect on the myeloma cells, by changing the bone marrow microenvironment may also play a anti-myeloma role.
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