摘要
目的探讨前B细胞白血病同源盒基因3（PBX3）基因小干扰RNA（siRNA）对脑胶质瘤细胞活力、凋亡的影响及其机制。方法参照Lipofectamine^TM2000说明，将PBX3的特异性siRNA（PBX3-siRNA组）转染人脑胶质瘤U87细胞，同时转染阴性对照siRNA（阴性组），并设置空白组，转染48h，Westernblot检测p38丝裂原活化蛋白激酶（p38MAPK）、磷酸化的p38丝裂原活化蛋白激酶（p-p38MAPK）、p53、含半胱氨酸的天冬氨酸蛋白水解酶3（Caspase．3）和活化的含半胱氨酸的天冬氨酸蛋白水解酶3（Cleaved Caspase-3）的蛋白表达，流式细胞术检测细胞凋亡率。噻唑蓝（Mrrr）检测PBX3．siRNA转染24、48、72h的细胞活力。结果转染PBX3．siRNA的U87细胞PBX3表达明显低于空白组（相对表达量分别为0．096±0．010、0．467±0．051，P=0．000）。与空白组比较，PBX3-siRNA组在24～72h的细胞活力均明显降低（24、48、72h细胞活力分别为0．336±0．034、0．549±0．057、0．671±0．073，P24h=0．001，P48h=0．004，P72h=0．004）。PBX3-siRNA组较空白组在48h的细胞凋亡率显著升高[凋亡率分别为（21．21±1．34）％，（4．71±0．32）％；P=0．000]。p-p38（0．031±0．006）蛋白表达显著降低，p53（0．388±0．041）和Cleaved-Caspase-3（0．205±0．018）蛋白表达显著升高（Pp-p38=0．000，Pp53=0．000，Pckaved-Caspase=0．000）。3组间p38和Caspase-3蛋白表达差异无统计学意义（Pp38=0．987，PCaspase-3=0．871）。结论PBX3基因siRNA可抑制脑胶质瘤细胞活力，诱导细胞凋亡，其机制可能是下调p38MAPK信号通路。
Objective To investigate the effect of pre - B - cell leukemia homeobox 3 （ PBX3 ） gene small interfering RNA （siRNA） on the viability and apoptosis of glioma cells. Methods According to LipofectamineTM 2000, the specific siRNA of PBX3 （ PBX3 - siRNA group） was transfeeted into human glioma U87 cells, and the negative control siRNA （negative group） was transfected, and the blank group was set up, cell was transfected for 48 h, the expression of p38, p - p38, p53, cysteinyl aspartate specific proteinase -3 （Caspase -3 ） and cleaved Caspase -3 protein were detected by Western blotting. The cell apoptosis rate was detected by flow cytometry. Methyl thiazol tetrazolium （MTY） was used to detect the cell viability after PBX3 - siRNA was transfeeted cells for 24, 48 and 72 h. Results The expression of PBX3 in U87 cells transfected with PBX3 - siRNA was significantly lower than that in blank group （ Protein rela- tive expression were O. 096 ±0. 010 and 0. 467±0. 051, P =0. 000）. Compared with the blank group, the cell viability in PBX3 - siRNA group （24, 48, 72 h were 0. 336± 0. 034, 0. 549 ± 0. 057, 0. 671±0. 073, P24 h = 0. 001, P48 h = 0. 004, P72 h = O. 004） decreased significantly PBX3 -siRNA was transfected for 24 - 72 h, the apoptosis rate increased significantly, the expression of p - p38 protein （0. 031 ± 0. 006） decreased significantly, and the expression of p53 （ 0. 388 ± 0. 041 ） and Cleaved - Caspase - 3 protein （0. 205 ±0. 018） increased significantly （Pp-ras =0. 000, Pp53 =0. 000, Pcleaved-Caspase-3 =0. 000）. There was no significant difference in the expression of p38 and Caspase - 3 between the three groups （ Pp38 = O. 987, Pcaspase-3 =0. 871 ）. Conclusion PBX3 gene siRNA can inhibit glioma cell viability and induce apoptosis, which may be related to downregulation of p38MAPK signal.
作者
李永明
赵亚军
秦涛
Li Yongming;Zhao Yajun;Qin Tao(Department of Neurosurgery,Henan Provincial People＇s Hospital,Zhengzhou 450003,Chin;Department of Hepatobiliary surgery,Henan Provincial People＇s Hospital,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2018年第9期1687-1690,共4页
Chinese Journal of Experimental Surgery
