Different pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state

doi:10.3969/j.issn.2095-4344.2014.37.006

Abstract

Abstract:

BACKGROUND: Recent studies have found that bone marrow mesenchymal stem cells play an important role in the development of multiple myeloma, especially for angiogenesis, but whether the pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state is different is yet unclear.
OBJECTIVE: To investigate the difference in the pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state.
METHODS: Bone marrow samples were extracted from 13 patients with multiple myeloma before treatment and after four therapeutic cycles to isolate bone marrow mesenchymal stem cells. ELISA assay was used to detect levels of vascular endothelial growth factor, hepatocyte growth factor and basic fibroblast growth factor in the supernatant of bone marrow mesenchymal stem cells. The proliferative ability and movement of human umbilical vein endothelial cells cultured in the supernatant of bone marrow mesenchymal stem cells were detected by MTT assay and Transwell assay, respectively. The pro-angiogenesis ability of bone marrow mesenchymal stem cells was measured by Matrigel in vitro.
RESULTS AND CONCLUSION: The levels of vascular endothelial growth factor, hepatocyte growth factor and basic fibroblast growth factor in the cell supernatant were significantly higher in active myeloma than those in remissive myeloma (P < 0.05). The supernatant of bone marrow mesenchymal stem cells of active myeloma more significantly promoted the proliferation, migration and blood vessel formation of human umbilical vein endothelial cells than that of remissive myeloma in a dose-dependent manner (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells from active myeloma have stronger pro-angiogenesis ability than those from remissive myeloma.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: bone marrow, mesenchymal stem cells, multiple myeloma, cell movement

CLC Number:

R394.2

Li Xiang-li, Zhang Xiao-ying, Zang Li, Wang Xiao-fang. Different pro-angiogenesis ability of bone marrow mesenchymal stem cells in multiple myeloma in active and remissive state[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(37): 5935-5941.

Figures/Tables 1

2.1 患者特征收集天津市肿瘤医院化疗前后的13例多发性骨髓瘤患者，其中男9例，女4例；年龄51-62岁，中位年龄为56岁；根据国际分期系统，Ⅰ、Ⅱ、Ⅲ期患者分别为1例(7.7%)、2例(15.4%)、10例(76.9%)；根据免疫球蛋白分型，IgG，IgA，λ轻链型分别为9例，3例，1例。根据国际骨髓瘤工作组统一标准[13]，疗效评价分为完全缓解(Complete response，CR)、部分缓解(partial response，PR)、病情平稳(stable disease，SD)、疾病进展(Progressive disease，PD)，完全缓解+部分缓解和病情平稳+疾病进展分别为6例，7例。 2.2 流式细胞仪分析骨髓间充质干细胞的免疫表型多发性骨髓瘤在活动期(治疗前+前病情平稳+疾病进展)及缓解期(完全缓解+部分缓解)骨髓间充质干细胞的免疫表型基本相似。都表达 CD106，CD105，CD29，CD44，不表达CD11a，CD11b，CD14，CD31，CD34，CD45，HLA-DR (表1)。 2.3 ELISA检测血管生成因子水平多发性骨髓瘤活动期患者骨髓间充质干细胞培养上清中血管内皮生长因子、肝细胞生长因子、碱性成纤维细胞生长因子的质量(P < 0.05，表2)；血管生成因子质量浓度在治疗前和活动期之间差异无显著性意义(P > 0.05)。浓度明显高于缓解期 2.4 骨髓间充质干细胞上清液对人脐静脉内皮细胞活性及增殖影响用MTT实验测定骨髓间充质干细胞培养上清液在不同的体积下对人脐静脉内皮细胞生长的影响。结果显示，骨髓瘤患者骨髓间充质干细胞上清对脐静脉内皮细胞增殖作用呈剂量依赖性，活动期明显强于缓解期(P < 0.05)。且治疗前组与病情平稳+疾病进展组间差异无显著性意义(P > 0.05，图1)。 2.5 骨髓间充质干细胞上清液对内皮细胞迁移影响通过Transwell小室实验发现骨髓间充质干细胞上清液影响人脐静脉内皮细胞向基底膜迁移，迁移数量在骨髓瘤活动期明显大于缓解期；统计学结果为骨髓瘤活动期患者骨髓间充质干细胞上清对脐静脉内皮细胞迁移作用明显强于缓解期(P < 0.05，图2)。且治疗前组与病情平稳+疾病进展组间差异无显著性意义(P > 0.05)。 2.6 体外血管形成能力体外成血管实验显示，骨髓瘤患者骨髓间充质干细胞上清在体外具有明显成血管作用，活动期明显强于缓解期(P < 0.05)。骨髓间充质干细胞上清的体外成血管能力在治疗前组与病情平稳+疾病进展组间差异无显著性意义(P > 0.05，图3A)。此外，多发性骨髓瘤活动期骨髓间充质干细胞体外直接成管作用明显强于缓解期(P < 0.05，图3B)，且治疗前组与病情平稳+疾病进展组间差异无显著性意义(P > 0.05)。"

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