K252a对表达TrkB的NB细胞耐药性的影响  

The Influence of K252a on Drug Resistance of Neuroblastoma that Expresses TrkB

作　　者：张继红[1] 李爱敏[2] 滕绍春 张可仞中国医科大学第二临床学院小儿外科 张锦华[1] 

机构地区：[1]中国医科大学第二临床学院血液研究室,沈阳市110004 [2]山东省烟台市毓璜顶医院儿科 [3]辽宁省辽阳市中心医院检验科

出　　处：《中国肿瘤临床》2005年第20期1144-1147,共4页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金资助(编号:39470739);卫生部科学研究基金资助(编号:20122167);辽宁省教育厅科研基金资助(编号:202013122);辽宁省博士启动科研基金资助(编号:20041047)

摘　　要：目的:使用全反式维甲酸(ATRA)诱导SH-SY5YNB(SY5Y)细胞的TrkB表达,加入BDNF激活TrkB/BDNF信号传导通路使TrkB磷酸化,然后用特异性酪氨酸激酶抑制剂K252a阻断该通路,观察NB细胞对化疗药物—顺铂(CDDP)敏感性的变化。方法:用ATRA诱导SY5Y细胞的TrkB高表达后,用BDNF、CDDP和TrkB信号传导通路的特异性阻断剂—K252a处理,MTT法检测细胞的存活率变化;流式细胞技术(FCM)检测细胞凋亡率;应用免疫组化法检测细胞磷酸化TrkB(p-TrkB)的表达水平。结果:ATRA能特异性诱导TrkB的表达,加入BDNF后可激活TrkB/BDNF信号传导通路并能引起TrkB的自动磷酸化;K252a能够阻断TrkB的磷酸化和TrkB/BDNF信号传导通路;经CDDP处理的SY5Y细胞的存活率下降、凋亡率上升。结论:阻断TrkB/BDNF信号传导通路可提高共表达TrkB/BDNF的NB细胞的化疗敏感性。因此,应用TrkB的抑制剂治疗NB有望提高该病的临床治愈率。Objection： To use all-trans-retinoic （ATRA） to induce the expression of TrkB in neuroblastoma cell line-SH-SY5Y（SY5Y）, to add BDNF to causes autophosphorylation of TrkB （p-TrkB） and activate TrkB/BDNF signal transudation, than to observe the change of NB cells＇ sensibility to chemotherapy drug, i.e. cisplatin （CDDP） before and after the blocking of TrkB receptor pathway by specific tyrosin kinase inhibitor K252a, which can block TrkB receptor pathway. Methods： ATRA was used to induce the expression of TrkB of SY5Y cells, and the addition of BDNF, CDDP, and K252a; MTT assay was performed to examination the changing of cell survival rate. Apoptosis rate was measured by flow cytometry （FCM）. Immunohistochemistry was employed to measure the expression of the autophosphorylating TrkB. Results： The expression of TrkB could specially be induced by ATRA and TrkB-BDNF signal pathway could be activated. TrkB was autophosphorylated after the addition of BDNF. The autophosphorylation of TrkB and TrkB/BDNF signal pathway could be blocked by K252a, the survival rate of SY5Y after treating with CDDP was decreased and the apoptosis rate was increased. Conclusion： The blockage of TrkB/BDNF signal pathway can increase the sensibility of chemotherapy in co-expressing TrkB and BDNF NB. Therefore treatment with inhibitors of TrkB receptors may provide potential novel therapeutic options in NB.

关 键 词：神经母细胞瘤 TrkB/BDNF信号传导通路 化疗耐药 K252A 

分 类 号：R73-3[医药卫生—肿瘤]