人多发性骨髓瘤RPMI 8226细胞条件培养液诱导破骨前体细胞的分化成熟  

Effect of conditioned medium of human multiple myeloma RPMI 8226 cells on osteoclastogenesis of RAW264.7 cells

作　　者：汤丽苑[1] 徐钰[2] 陈瑾[2] 刘新华[3] 俞康[1] 

机构地区：[1]温州医学院附属第一医院血液科 [2]温州医学院 [3]温州医学院血液肿瘤与移植免疫研究所,浙江温州325000

出　　处：《中国病理生理杂志》2012年第4期649-654,共6页Chinese Journal of Pathophysiology

基　　金：温州医学院5010重大项目子课题(No.XNK05005)

摘　　要：目的:探讨人多发性骨髓瘤RPMI 8226细胞条件培养液对破骨前体细胞RAW264.7的影响。方法:Western blotting检测可溶性核因子κB受体激活剂配体(soluble receptor activator of NF-κB ligand,sRANKL)的蛋白表达。抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)染色观察RAW264.7细胞的分化成熟情况。RT-PCR法检测RAW264.7细胞TRAP和cathepsin K mRNA的表达。结果:Western blotting法证实RPMI8226细胞条件培养液中含有sRANKL。TRAP染色发现RPMI 8226细胞条件培养液能诱导RAW264.7细胞分化为TRAP阳性的多核成熟破骨细胞。人抑制性RANKL单克隆抗体拮抗30%RPMI 8226细胞条件培养液中sRANKL的作用,且具有剂量依赖性。30%RPMI 8226细胞条件培养液能刺激RAW264.7细胞上调TRAP和cathepsin KmRNA表达。结论:人多发性骨髓瘤细胞RPMI 8226条件培养液中的sRANKL具有促RAW264.7破骨前体细胞分化成TRAP阳性的多核成熟破骨细胞的生物活性。人抑制性RANKL单克隆抗体阻断30%RPMI 8226细胞条件培养液中sRANKL的诱导分化作用,且具有浓度依赖性。AIM： To investigate the osteoclastogenic effect of conditioned medium of human multiple myeloma RPMI 8226 cells on preosteoclast RAW264.7 cells.METHODS： The protein expression of soluble receptor activator of NF-κB ligand（sRANKL） was detected by Western blotting.The morphological changes of RAW264.7 cells were observed after tartrate-resistant acid phosphatase（TRAP） staining.The mRNA expression of TRAP and cathepsin K was evaluated by RT-PCR.RESULTS： The result of Western blotting showed that conditioned medium of RPMI 8226 cells contained sRANKL.RPMI 8226 cell conditioned medium induced RAW264.7 cells to differentiate into TRAP-positive multinuclear osteoclasts.Human neutralized RANKL monoclonal antibody suppressed the differentiation of preosteoclasts in a dose-dependent manner,which was induced by 30% RPMI 8226 cell conditioned medium.RPMI 8226 cell conditioned medium increased the mRNA expression of TRAP and cathepsin K in RAW264.7 cells.CONCLUSION： The sRANKL in conditioned medium of human multiple myeloma RPMI 8226 cells has the bioactivity to induce preosteoclast RAW264.7 cells to differentiate into TRAP-positive multinuclear osteoclasts.Human neutralized RANKL monoclonal antibody suppresses the differentiation of preosteoclasts induced by sRANKL in a dose-dependent manner.

关 键 词：多发性骨髓瘤 破骨细胞 核因子κB受体激活剂配体 

分 类 号：R733.3[医药卫生—肿瘤]