多发性骨髓瘤患者成骨细胞培养及调控  

Culture and regulation of osteoblasts in multiple myeloma patients

作　　者：高珊[1] 付蓉[1] 刘惠[1] 刘鸿[1] 宋嘉[1] 王化泉[1] 王国锦[1] 邢莉民[1] 邵宗鸿[1] 

机构地区：[1]天津医科大学总医院血液科,300052

出　　处：《中华血液学杂志》2014年第3期202-206,共5页Chinese Journal of Hematology

基　　金：天津市抗癌重大专项攻关计划（12ZCDZSY18000）;中国医师协会项目（20111207、20090109）

摘　　要：目的研究多发性骨髓瘤（MM）患者骨髓来源成骨细胞的增殖分化和成骨潜能，比较蛋白酶体抑制剂硼替佐米及MM患者血清对成骨细胞的影响。方法以20例MM患者和10名健康正常人为研究对象，采用骨髓培养法从骨髓中培养并纯化出成骨细胞，进行成骨细胞计数；成骨细胞传代培养4周后，行Von Kossa’s钙染色，进行成骨细胞钙结节计数；在MM患者组及正常对照组成骨细胞中分别加入硼替佐米及MM患者血清，对比其细胞生物学特性的变化。采用流式细胞术检测MM患者成骨细胞抗体CD34、CD138、CD45的表达，采用ELISA法测定血清IL-7水平，采用RT-PCR法检测各组培养细胞中骨形成蛋白2（BMP2）的表达。结果①成骨细胞计数（单位为X104／m1）：MM患者组（6．3±1．5）较正常对照组（8．2±2．6）减少（P〈0．05）；MM患者组中，加硼替佐米组（8．9±2．1）较不加硼替佐米组（7．8±3．0）明显增多（P〈0．05），但加入MM血清与否差异无统计学意义[（5．9±1．6）对（6．3±1．5），P〉0．05]；正常对照组中，加入MM血清组（7．4±1．1）较不加MM血清组（8．2±2．6）有所减少（P〈0．05）。②成骨细胞钙结节计数：MM患者组中，加硼替佐米组（8．8-4-1．9）多于不加硼替佐米组（6．1±1．6）（P〈0．05），但少于正常对照组（15．8±2．2）（P〈0．05）。③各组成骨细胞均不表达CDl38、CD45、CD34。④MM患者组血清IL-7水平（2．07±0．71）高于正常对照组（1．62±0．15）（P〈0．05）。⑤加硼替佐米MM患者组及正常对照组均可表达BMP2mRNA，不加硼替佐米的MM患者组BMP2mRNA无明显表达。结论骨髓培养法可用于体外培养MM患者成骨细胞。与正常对照者比较，MM患者成骨细胞的增殖分化及成骨能力明显减弱。硼替佐米有促进成骨细胞增殖分化及成骨的能力；MM患者血清可抑制成骨细胞的增殖�Objective To investigate the biological characteristics ofosteoblasts cultured in vitro from bone marrow （BM） of multiple myeloma （MM） patients and to explore their generation and osteogenic potential. Effects of some factors such as bortezom ib and MM patient serum on the osteoblasts were observed. Methods Twenty MM patients and 10 healthy donors as controls were enrolled in this study. Osteoblasts from MM patients＇ BM were cultured in vitro. The generation and osteogenic potential of osteoblasts from MM patients and normal subjects were compared. The changes of their osteogenic potential and biological characteristics were observed. The antigens （CD34, CD138, CD45） on osteoblasts were cwalyzed by flow cytometry. The levels of IL-7 were measured by ELISA. The BMP2 mRNAs were measured by RT-PCR. Results Osteoblasts from MM patients＇ BM could be cultured in vitro. The quantity of osteoblasts from MM patients （6.3 ± 1.5） was less than normal subjects （8.2± 2.6） （P〈0.05）. The osteoblasts cultured with MM patient serum （7.4±1. 1 ）were less than those without patient serum （8.2~2.6） （P〈0.05）. Bortezomib increased those from MM patients after 6 days culture （8.9~2.1 vs 6.3±1.5） （P〈 0.05）. Von Kossa staining showed that there were more calcium depositions in MM osteoblasts with bortezomib （8.8 ± 1.9） than those without bortezomib （6.1 ± 1.6） （P〈0.05）, but less than those from normal controls （15.8±2.2）（P〈0.05）. CD138, CD45, CD34 were not detected on the cultured cells. The level of IL-7 in MM patients＇ serum（2.07±0.71 ） was higher than that in normal controls（1.62±0.15） （P〈0.05）. The expression of BMP2 mRNA was seen in the normal osteoblasts and MM patients＇ osteoblasts culured with bortezomib, but not be seen in those without bortezomib. Conclusion Osteoblasts could be cultured in vitro from MM patients＇ BM. The proliferation and osteogenic potential of osteoblasts from MM patients were decreased. Bortezomib was a

关 键 词：多发性骨髓瘤 成骨细胞 原代细胞培养 硼替佐米 

分 类 号：R733.3[医药卫生—肿瘤]