表达脑源性神经营养因子人多发性骨髓瘤小鼠模型建立.doc

表达脑源性神经营养因子人多发性骨髓瘤小鼠模型建立 　 作者：王雅丹，胡豫，张璐，黄靖，孙春艳 【摘要】 过去的研究证实脑源性神经营养因子（BDNF）在体外具有促进多发性骨髓瘤（MM）细胞增殖并诱导MM血管新生的能力。本研究探讨BDNF/TrkB途径是否为治疗MM的潜在靶点，并比较两种途径建立人MM NOD/SCID小鼠模型的优缺点，为深入探索治疗MM的新靶点奠定基础。选择糖尿病抵抗/重症联合免疫缺陷（NOD/SCID）小鼠，通过皮下注射或尾静脉注射人骨髓瘤细胞株RPMI8226建立两种异体移植动物模型。观察荷瘤后小鼠的生长状态，测量皮下瘤块的体积；荷瘤后3周，每周经眼眶后静脉丛采血，检测血清中人源λ轻链含量、Ca2+浓度和血浆中人源BDNF浓度，并计数红细胞；小鼠死后，采用组织学方法观察瘤细胞的形态特征，流式细胞术检测小鼠外周血和骨髓中人源性CD38＋细胞比例，采用计算机X线数字摄影观察小鼠全身骨密度的改变和骨质破坏情况。结果表明：皮下注射模型的成瘤率高（5/5），具有多种与浆细胞瘤相似的病理学特征，但其骨髓中未检测到MM细胞，血清中钙离子浓度不高，M蛋白浓度升高不明显且未发现溶骨性损害的组织学和影像学证据。尾静脉注射模型成瘤率相对较低（4/7），骨髓中可检测到呈浸润生长的人CD38＋细胞；而且在荷瘤3周后，血清中即可检测到人源M蛋白；随着肿瘤的生长，M蛋白水平、钙离子浓度逐渐升高，并有溶骨性损害的影像学证据。两种模型血浆中人源BDNF的水平亦逐渐升高，9周时浓度分别为（73±11）pg/ml和（105±18）pg/ml。结论：本研究成功建立了两种高表达BDNF的MM荷瘤NOD/SCID小鼠模型，两种模型相互结合应用，为探索MM治疗的新靶点BDNF/TrkB提供了合适的动物模型。 【关键词】 多发性骨髓瘤 Establishment of Multiple Myeloma Mouse Models Expressing Brain Derived Neurotrophic Factor Abstract Previous studies have demonstrated the effects of brainderived neurotrophic factor (BDNF) on promoting proliferation of multiple myeloma (MM) cells and inducing angiogenesis in MM in vitro. This study was aimed to further explore whether BDNF/TrkB pathway is a potential therapeutic target in MM, and to elucidate the advantages and disadvantages of two ways developed for human myeloma xenograft in animal models. The models of xenograft tumors were established in the nonobese diabetic /severe combined immunodeficiency (NOD/SCID) mice by subcutaneous or intravenous injection of human myeloma cell line RPMI8226. Mice were monitored daily for life state, and the volume of subcutaneous tumors were measured after inoculation. 3 weeks after inoculation, red blood cell counts, BDNF level in plasma, human λ light chain and calcium level in serum of NOD/SCID were detected every two weeks. The histological and cytological examinations were performed to observe pathological features of tumors. Using flow cytometry to observe the expression of human CD38+ cell in murine blood and bone marrow. The changes of bone density and