Mahtouk Karène, Hose Dirk, Raynaud Pierre, Hundemer Michael, Jourdan Michel, Jourdan Eric, Pantesco Veronique, Baudard Marion, De Vos John, Larroque Marion, Moehler Thomas, Rossi Jean-Francois, Rème Thierry, Goldschmidt Hartmut, Klein Bernard
Institute of Research in Biology, Centre Hospitalier Universitaire Montpellier, 99 rue Puech Villa, 34197 Montpellier Cedex 5, France.
Blood. 2007 Jun 1;109(11):4914-23. doi: 10.1182/blood-2006-08-043232. Epub 2007 Mar 5.
The heparan sulfate (HS) proteoglycan, syndecan-1, plays a major role in multiple myeloma (MM) by concentrating heparin-binding growth factors on the surface of MM cells (MMCs). Using Affymetrix microarrays and real-time reverse transcriptase-polymerase chain reaction (RT-PCR), we show that the gene encoding heparanase (HPSE), an enzyme that cleaves HS chains, is expressed by 11 of 19 myeloma cell lines (HMCLs). In HSPE(pos) HMCLs, syndecan-1 gene expression and production of soluble syndecan-1, unlike expression of membrane syndecan-1, were significantly increased. Knockdown of HPSE by siRNA resulted in a decrease of syndecan-1 gene expression and soluble syndecan-1 production without affecting membrane syndecan-1 expression. Thus, HPSE influences expression and shedding of syndecan-1. Contrary to HMCLs, HPSE is expressed in only 4 of 39 primary MMC samples, whereas it is expressed in 36 of 39 bone marrow (BM) microenvironment samples. In the latter, HPSE is expressed at a median level in polymorphonuclear cells and T cells; it is highly expressed in monocytes and osteoclasts. Affymetrix data were validated at the protein level, both on HMCLs and patient samples. We report for the first time that a gene's expression mainly in the BM environment (ie, HSPE) is associated with a shorter event-free survival of patients with newly diagnosed myeloma treated with high-dose chemotherapy and stem cell transplantation. Our study suggests that clinical inhibitors of HPSE could be beneficial for patients with MM.
硫酸乙酰肝素(HS)蛋白聚糖syndecan-1通过将肝素结合生长因子集中在多发性骨髓瘤(MM)细胞(MMCs)表面,在MM中发挥重要作用。使用Affymetrix微阵列和实时逆转录聚合酶链反应(RT-PCR),我们发现编码硫酸乙酰肝素酶(HPSE)(一种切割HS链的酶)的基因在19个骨髓瘤细胞系(HMCLs)中的11个中表达。在HSPE阳性的HMCLs中,syndecan-1基因表达和可溶性syndecan-1的产生,与膜syndecan-1的表达不同,显著增加。通过小干扰RNA(siRNA)敲低HPSE导致syndecan-1基因表达和可溶性syndecan-1产生减少,而不影响膜syndecan-1的表达。因此,HPSE影响syndecan-1的表达和脱落。与HMCLs相反,HPSE仅在39个原发性MMC样本中的4个中表达,而在39个骨髓(BM)微环境样本中的36个中表达。在后者中,HPSE在多形核细胞和T细胞中以中等水平表达;在单核细胞和破骨细胞中高表达。Affymetrix数据在蛋白水平上在HMCLs和患者样本中均得到验证。我们首次报道,一个主要在BM环境中表达的基因(即HSPE)与接受大剂量化疗和干细胞移植的新诊断骨髓瘤患者较短的无事件生存期相关。我们的研究表明,HPSE的临床抑制剂可能对MM患者有益。