Identification of the source of elevated hepatocyte growth factor levels in multiple myeloma patients.
Identification of the source of elevated hepatocyte growth factor levels in multiple myeloma patients.
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DOI：
10.1186/2050-7771-2-8
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发表时间：
2014-04-09
影响因子：
11.1
通讯作者：
Sundan A
中科院分区：
文献类型：
--
作者：
Rampa C;Tian E;Våtsveen TK;Buene G;Slørdahl TS;Børset M;Waage A;Sundan A
肝细胞生长因子（HGF）是一种多用途细胞因子，可导致癌细胞增殖、迁移和转移。在多发性骨髓瘤（MM）患者中，它是骨髓中丰富的成分。50%的患者HGF水平升高，并伴有预后不良。我们的目的是研究其在骨髓瘤中的来源。实时荧光定量PCR法测定健康人及骨髓瘤患者骨髓活检组织中HGF mRNA水平。通过微阵列分析，对健康个体和MM患者骨髓中分离的CD138+细胞进行HGF基因表达谱分析。采用酶联免疫吸附试验（ELISA）测定MM患者外周血HGF蛋白浓度。采用荧光原位杂交（FISH）和HGF基因启动子DNA测序技术检测CD138+细胞的细胞遗传学状态。ELISA法测定人骨髓瘤细胞系与骨髓基质细胞共培养时HGF的分泌情况。骨髓核心活检和CD138+细胞中的HGF基因表达谱显示骨髓瘤患者HGF mRNA水平升高。活组织组织中HGF mRNA水平与骨髓瘤细胞相关。血清中HGF蛋白水平的定量也与相应患者CD138+细胞中HGF mRNA水平相关。细胞遗传学分析显示，骨髓瘤细胞克隆的HGF拷贝数在1 ~ 3个拷贝之间。HGF拷贝数与HGF mRNA水平无相关性。人骨髓瘤细胞系ANBL-6和JJN3与骨髓基质细胞或HS-5细胞系共培养可显著增加HGF的分泌。我们在这里表明，在骨髓瘤患者中，HGF主要由恶性浆细胞产生，这些细胞的HGF产生可能受到骨髓微环境的支持。考虑到血清和血浆HGF水平升高预示预后不良，这些发现对骨髓瘤克隆产生大量HGF的患者尤为重要。
Hepatocyte growth factor (HGF) is a pleiotropic cytokine which can lead to cancer cell proliferation, migration and metastasis. In multiple myeloma (MM) patients it is an abundant component of the bone marrow. HGF levels are elevated in 50% of patients and associated with poor prognosis. Here we aim to investigate its source in myeloma. HGF mRNA levels in bone marrow core biopsies from healthy individuals and myeloma patients were quantified by real-time PCR. HGF gene expression profiling in CD138+ cells isolated from bone marrow aspirates of healthy individuals and MM patients was performed by microarray analysis. HGF protein concentrations present in peripheral blood of MM patients were measured by enzyme-linked immunosorbent assay (ELISA). Cytogenetic status of CD138+ cells was determined by fluorescence in situ hybridization (FISH) and DNA sequencing of the HGF gene promoter. HGF secretion in co-cultures of human myeloma cell lines and bone marrow stromal cells was measured by ELISA. HGF gene expression profiling in both bone marrow core biopsies and CD138+ cells showed elevated HGF mRNA levels in myeloma patients. HGF mRNA levels in biopsies and in myeloma cells correlated. Quantification of HGF protein levels in serum also correlated with HGF mRNA levels in CD138+ cells from corresponding patients. Cytogenetic analysis showed myeloma cell clones with HGF copy numbers between 1 and 3 copies. There was no correlation between HGF copy number and HGF mRNA levels. Co-cultivation of the human myeloma cell lines ANBL-6 and JJN3 with bone marrow stromal cells or the HS-5 cell line resulted in a significant increase in secreted HGF. We here show that in myeloma patients HGF is primarily produced by malignant plasma cells, and that HGF production by these cells might be supported by the bone marrow microenvironment. Considering the fact that elevated HGF serum and plasma levels predict poor prognosis, these findings are of particular importance for patients harbouring a myeloma clone which produces large amounts of HGF.