DAPT与K252a联用体外抑制人脑胶质瘤U251MG细胞的生长  

Combined use of DAPT and K252a inhibits in vitro growth of human glioma U251MG cells

作　　者：陈普建 熊二梦[2] 张严[2] 龚爱华[2] 

机构地区：[1]南通市第四人民医院神经外科,江苏省226005 [2]江苏大学基础医学与医学技术学院

出　　处：《江苏医药》2014年第15期1746-1749,I0001,共5页Jiangsu Medical Journal

摘　　要：目的探讨Notch信号抑制剂(2S)-N-N-(3,5-氟苯乙酰基)-L-丙氨酰-2-苯基甘氨酸叔丁酯(DAPT)与神经营养素受体高亲和力的酪氨酸激酶受体家族(Trks)阻断剂K252a联用对人脑胶质瘤U251MG细胞的抑制作用。方法应用DAPT和K252a单独或联用处理U251MG细胞,噻唑蓝比色实验检测细胞的活性,流式细胞术检测细胞凋亡情况,Western blot检测p-TrkA、JNK、p-JNK、Caspase-3、Caspase-9、Bax、Bcl-2和Bcl-xL等蛋白的表达。结果 DAPT和K252a均能呈时间与剂量依赖性抑制体外胶质瘤细胞的活性。K252a200、100和50nmol/L与DAPT 2.5μmol/L联合用药24、48和72h的细胞活力均低于单独使用K252a或DAPT(P<0.05)。两药联用可以增加sub-G1期细胞,即促进细胞凋亡。与K252a100nmol/L或DAPT 2.5μmol/L单用比较,K252a100nmol/L与DAPT 2.5μmol/L联用降低了p-TrkA的量,对于JNK的表达基本没影响,但可增加p-JNK的表达。可活化Caspase-3和Caspase-9,同时减少Bcl-xL的表达,但对Bax和Bcl-2的表达基本没影响。结论 DAPT能抑制U251MG的细胞活性。联合DAPT用药能增强K252a对人脑胶质瘤细胞U251MG的毒性。Objective To explore the in vitro inhibition effect of combined use of DAPT （gamma secretase inhibitor） and K252a（Trks inhibitor） on the growth of human glioma U251MG cells. Methods The viability of U251MG cells was detected using MTT assay. Apoptosis levels were examined by FACS, and the expressions of p-TrkA, JNK, p-JNK, Caspase-3, Caspase-9, Bax, Bcl-2, and Bcl-xL were determined by Western blot. Results DAPT and K252a inhibited in vitro the growth of U251MG cells in the dose- and time-dependent manners. The cell viability was significantly decreased in combined use of DAPT and K252a compared with K252a or DAPT used alone at different time points of 24,48 and 72 hours. Apoptosis levels were increased in U251MG cells treated with combined use of DAPT and K252a. Furthermore, the expression levels of apoptosis-related proteins Caspase-3, Caspase-9, and p-JNK were obviously increased, while the expressions of Bcl-xL and p-TrkA were decreased when combined use of DAPT and K252a. Conclusion DAPT can inhibit in vitro the viability of U251MG cells. Combined use of DAPT may enhance toxic effect of K252a on human glioma U251MG cells.

关 键 词：DAPT K252A 胶质瘤 

分 类 号：R737[医药卫生—肿瘤]