血管内皮生长因子对骨髓瘤骨髓基质细胞分泌白细胞介素6的影响  

Influence of vascular endothelial growth factor on interleukin-6 secretion in myeloma bone marrow stromal cells

作　　者：王晓波[1] 许咏青[2] 

机构地区：[1]大连医科大学附属二院血液内科,辽宁省大连市116027 [2]大连市儿童医院内科,辽宁省大连市116027

出　　处：《中国组织工程研究与临床康复》2008年第25期4899-4902,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景：研究表明骨髓瘤细胞和骨髓基质细胞黏附后，可促进多种细胞因子如血管内皮生长因子、白细胞介素6的分泌，这些因子在促进肿瘤生长、浸润中起到关键作用。目的：观察血管内皮生长因子对多发性骨髓瘤患者骨髓基质细胞分泌白细胞介素6的影响。设计、时间及地点：对照观察，于2005—04／2007—04在大连医科大学附属二院血液实验室完成。对象：所有骨髓标本来源于大连医科大学附属二院血液科住院患者，多发性骨髓瘤患者18例，缺铁性贫血患者8例的骨髓为正常对照。血管内皮生长因子为R＆D公司产品。方法：上述对象每例抽取骨髓5mL，贴壁法分离培养骨髓基质细胞。当细胞贴满瓶壁后传代，取2～3代细胞进行实验。分别向骨髓瘤骨髓基质细胞、正常骨髓基质细胞的悬液中加入含10％胎牛血清的IMDM培养液，按1×10^0L^-1密度接种，48h后收集上清液待测。此外，分别以10，50，100μg/L血管内皮生长因子作用骨髓瘤骨髓基质细胞48h，另以100μg／L血管内皮生长因子分别作用骨髓瘤骨髓基质细胞24，48，72h，收集上清液待测。主要观察指标：倒置显微镜下观察细胞生长情况，流式细胞仪检测细胞表型。ELISA法检测两种骨髓基质细胞培养上清液中的白细胞介素6浓度。ELISA法检测血管内皮生长因子作用于骨髓瘤骨髓基质细胞对分泌白细胞介素6的影响。结果：26例骨髓标本均进入结果分析。①体外培养的骨髓基质细胞贴壁生长，平均4周融合为单层，传代后表现为成纤维细胞形态，其表型为CD54^＋，CD33^-，CD31^-。②骨髓瘤骨髓基质细胞上清液中白细胞介素6的含量明显高于正常骨髓基质细胞（t=6．17，P〈0．05）。③分别以10，50，100μg／L血管内皮生长因子作用骨髓瘤骨髓基质细胞48h后，其上清液中白细胞介素6的含量逐渐升高，3种浓度间比较差BACKGROUND： Myeloma cell and bone marrow stromal cell adherence can promote the secretion of many cytokines such as vascular endothelial growth factor （VEGF） and interleukin-6, which can play an important role in tumor growth and infiltration. OBJECTIVE： To investigate the VEGF effects on interleukin-6 secretion in multiple myeloma patients. DESIGN, TIME AND SETTING： The control observation was performed at the Laboratory of Hematology, Second Affiliated Hospital, Dalian Medical University from April 2005 to April 2007. PARTICIPANTS： Bone marrow samples were obtained from 18 patients with multiple myeloma and 8 patients with iron deficiency anemia （served as normal control） at the Department of Hematology, Second Affiliated Hospital, Dalian Medical University. VEGF was purchased from R＆D, USA. METHODS： 5 mL bone marrow was collected from each above-mentioned patient. Bone marrow stromal cells were harvested by adherence. After confluence, the second and third passages of cells were used in this study. Myeloma bone marrow stromal cells and normal bone marrow stromal cells were separately incubated in IMDM medium containing 10% fetal bovine serum at a density of 1 × 10^9 L^-1. 48 hours later, supematant was collected. Myeloma bone marrow stromal cells were treated with 10, 50, 100 μ g/L VEGF for 48 hours. Moreover, myeloma bone marrow stromal cell were treated with 100 μg/L VEGF for 24, 48 and 72 hours, separately. Supematant was collected for detection. MAIN OUTCOME MEASURES： Cell growth was observed under an inverted microscope. Cell phenotype was detected by flow cytometry. Interleukin-6 levels in supematant of two kinds of cells were measured by enzyme-labeled immunosorbent assay. VEGF effects on ihterleukin-6 in myeloma bone marrow stromal cells were examined by enzyme-labeled immunosorbent assay. RESULTS： Totally 26 bone marrow samples were included in the final analysis. In vitro cultured bone marrow stromal cells adhered to the flask, and confluence into a monolayer ever

关 键 词：骨髓基质细胞 血管内皮生长因子 白细胞介素6 多发性骨髓瘤 

分 类 号：R394.2[医药卫生—医学遗传学]