脑源性神经营养因子对人骨髓瘤细胞血管生成的影响

[Effect of brain-derived neurotrophic factor in human myeloma cells on angiogenesis].

作者信息

Hu Yu, Wu Tao, Wang Ya-Dan, He Wen-Juan

机构信息

Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2005 Oct;26(10):602-6.

PMID:16532969

Abstract

OBJECTIVE

To investigate the expression of brain-derived neurotrophic factor (BDNF) in multiple myeloma (MM) cells and the correlation between BDNF and MM angiogenesis.

METHODS

The expressions of BDNF mRNA transcripts and protein in MM cell lines (RPMI 8226, KM3) were determined by RT-PCR and Western blot, respectively, BDNF levels in culture supernatant by enzyme-linked immunosorbent assay. Proliferation of human umbilical vein endothelial cells (HUVEC) mixed with MM culture medium at different concentrations was examined by MTT assay. The effects of MM culture medium on HUVEC migration and tube formation were studied by modified Boyden chamber assay and tube formation assay, respectively.

RESULTS

BDNF was expressed in and secreted by MM cell lines RPMI 8226 and KM3. BDNF concentrations in culture supernatants were within the range of its biological activity. MM culture medium induced a concentration-dependent proliferation of HUVEC. The number of HUVEC at a concentration of 50% KM3 culture medium and at full KM3 culture medium were (1.85 +/- 0.23)-fold and (2.16 +/- 0.29) -fold increase, respectively (P <0.05), compared with that of control. The proliferative activity of HUVEC was reduced on the addition of BDNF antibody to the culture medium. MM culture medium also stimulated the migration and differentiation of HUVEC in vitro, the chemotactic index of HUVEC at a concentration of 50% KM3 culture medium and at full KM3 culture medium were 1.85 +/- 0.23 and 2.16 +/- 0.29, respectively (P < 0.05). Full KM3 culture medium also stimulated capillary-like tube formation in HUVEC (P <0.01), and addition of anti-human BDNF antibody neutralized these effects significantly.

CONCLUSION

MM cell lines expressed and secreted biologically active BDNF, which may be involved, at least in part, in MM angiogenesis.

摘要

目的

研究脑源性神经营养因子（BDNF）在多发性骨髓瘤（MM）细胞中的表达及其与MM血管生成的相关性。

方法

分别采用逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测MM细胞系（RPMI 8226、KM3）中BDNF mRNA转录本和蛋白质的表达，采用酶联免疫吸附测定法检测培养上清液中BDNF水平。采用MTT法检测不同浓度MM培养基与人脐静脉内皮细胞（HUVEC）共培养时HUVEC的增殖情况。分别采用改良Boyden小室法和体外成管实验研究MM培养基对HUVEC迁移和成管的影响。

结果

BDNF在MM细胞系RPMI 8226和KM3中表达并分泌。培养上清液中BDNF浓度在其生物活性范围内。MM培养基诱导HUVEC呈浓度依赖性增殖。与对照组相比，50% KM3培养基浓度组和完全KM3培养基浓度组的HUVEC数量分别增加了（1.85±0.23）倍和（2.16±0.29）倍（P<0.05）。向培养基中添加BDNF抗体可降低HUVEC的增殖活性。MM培养基还可刺激HUVEC在体外的迁移和分化，50% KM3培养基浓度组和完全KM3培养基浓度组的HUVEC趋化指数分别为1.85±0.23和2.16±0.29（P<0.05）。完全KM3培养基还可刺激HUVEC形成毛细血管样管腔（P<0.01），添加抗人BDNF抗体可显著中和这些作用。