神经营养因子酪氨酸激酶受体B对人永生化成骨细胞hFOB1.19恶性转化细胞体外侵袭力的影响  

Effect of neurotrophic tyrosine kinase receptor B on in vitro invasiveness of malignant transformed hFOB1.19 cells

作　　者：曾静[1] 温磊[1] 黄玉胜[1] 戴欢子[1] 郭乔楠[1] 

机构地区：[1]第三军医大学西南医院病理学研究所,重庆400038

出　　处：《第三军医大学学报》2010年第9期938-942,共5页Journal of Third Military Medical University

摘　　要：目的研究神经营养因子酪氨酸激酶受体B(neurotrophic tyrosine kinase receptor B,TrKB)对恶性转化人永生化成骨细胞hFOB1.19体外侵袭力的影响,探讨TrKB在骨肉瘤侵袭和转移机制中发挥的作用。方法RT-PCR和免疫荧光细胞染色检测hFOB1.19恶性转化细胞和SaOS-2骨肉瘤细胞中TrKB的mRNA和蛋白表达水平;进一步研究hFOB1.19恶性转化细胞中TrKB的功能,处理组hFOB1.19恶性转化细胞加入特异性酪氨酸激酶抑制剂K252a处理24h,非处理组加入DMSO作对照,倒置相差显微镜下观察细胞形态;Transwell侵袭实验检测细胞侵袭能力;actin免疫荧光细胞染色检测细胞微丝骨架。结果TrKB在hFOB1.19恶性转化细胞中mRNA和蛋白表达水平均明显高于SaOS-2骨肉瘤细胞(P<0.05);K252a处理组hFOB1.19恶性转化细胞和未处理组相比,细胞变圆、聚集甚至发生融合,侵袭能力明显减弱(P<0.01),微丝回缩变短,排列紊乱。结论人永生化成骨细胞hFOB1.19恶性转化细胞中TrKB的高表达可促进该细胞的体外高侵袭力;该受体被阻断后,可能通过改变细胞微丝骨架结构从而降低细胞的体外侵袭力;TrKB可能在骨肉瘤侵袭和转移机制中起着促进作用。Objective To study the effect of neurotrophic tyrosine kinase receptor B（TrKB） on in vitro invasiveness of malignant transformed hFOB1.19 cells and the role of TrKB in invasion and metastasis of osteosarcoma.Methods Expression of TrKB in malignant transformed hFOB1.19 cells and SaOS-2 osteosarcoma cells was detected by RT-PCR and immunofluorescence.Function of TrKB of malignant transformed hFOB1.19 cells was further studied.Malignant transformed hFOB1.19 cells were treated with specific tyrosine kinase inhibitor K252a for 24 h as a treatment group,and untreated malignant transformed hFOB1.19 cells into which DMSO was added served as a control group.Morphology of cells was observed under an inverted phase contrast microscope.Cell invasiveness was detected by Transwell assays.Microfilaments of cells were detected by actin immunofluorescence.Results The expression of TrKB was significantly higher in malignant transformed hFOB1.19 cells than in SaOS-2 osteosarcoma cells（P0.05）.The cells in treatment group became round,aggregated,and even fused compared to those in control group.The cell invasiveness was significantly lower in treatment group than in control group （P0.01）.The microfilaments of cells were shorter and more disordered in arrangement in treatment group than in control group.Conclusion High expression level of TrKB in malignant transformed hFOB1.19 cells can increase the in vitro invasiveness of such cells.Blockage of TrKB can decrease the in vitro invasiveness of malignant transformed hFOB1.19 cells by changing their microfilament structure.TrKB may play a positive role in invasion and metastasis of osteosarcoma.

关 键 词：神经营养因子酪氨酸激酶受体B K252A 骨肉瘤 微丝 侵袭 转移 

分 类 号：R341[医药卫生—基础医学] R730.23