葛根素与三氧化二砷协同作用通过Akt/p38途径促进人胶质瘤细胞凋亡  

Puerarin Synergizes with Arsenic Trioxide for the Apoptosis of Human Glioblastoma Cells through the Protein Kinase/p38 Mitogen-activated Protein Kinases Pathway

作　　者：羊轶驹[1] 孙振球[1] 

机构地区：[1]中南大学公共卫生学院社会医学与卫生管理系,长沙市410083

出　　处：《中国肿瘤临床》2012年第24期2059-2062,共4页Chinese Journal of Clinical Oncology

摘　　要：目的:研究葛根素(PRN)是否通过Akt/p38途径协同三氧化二砷(As_2O_3)促进人胶质瘤细胞的凋亡。方法:MTT检测细胞的存活率,流式细胞仪(FCM)技术检测细胞的凋亡状态,蛋白免疫印迹(Immunoblotting)检测细胞phosphorylated Akt和p38-MAPK,Cleaved Caspase-3蛋白的表达,PCR检测Caspase-3的mRNA的表达。结果:PRN协同As_2O_3降低人胶质瘤细胞U87的存活。与对照组相比,PRN(16μM)组,As_2O_3(2μM)组显著增加细胞内钙水平(1.13±0.015),(1.18±0.33)。此外,PRN能够协同As_2O_3增加细胞内钙(1.34±0.72),下调蛋白phosphorylated Akt,上调phosphorylated p38-MAPK和Cleaved Caspase-3蛋白及Cleaved Caspase-3 mRNA表达水平。结论:PRN协同As_2O_3增加胶质瘤细胞内钙水平,抑制细胞存活。此外,PRN联合As_2O_3下调phosphorylated Akt,增强phosphorylated p38-MAPK和Cleaved Caspase-3的表达,进而促进肿瘤细胞凋亡。PRN可能成为临床上辅助As_2O_3治疗肿瘤中的潜在的辅助治疗药物。Objective： To investigate whether puerarin （PRN） synergizes with arsenic trioxide （As203） in facilitating the apoptosis of human glioblastoma cell line U87 through the protein kinase （Akt）/p 38 mitogen-activated protein kinases （p38-MAPK） pathway. Methods： The 3-（4,5-deimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide technique was performed to detect cell survival. Flow cytometry was applied to calculate cell apoptosis, and immunoblot technique was used to verify the protein expressions of phosphorylated Akt, p38-MAPK, and cleaved caspase-3. In addition, caspase-3 mRNA levels were detected by real-time polymerase chain reaction. Results： PRN synergized with As2O3 to decrease the survival of human glioblastoma cell line U87. A single dosage each of PRN （16 uM; 1.13±0.015） and As2O3 （2 uM; 1.184-0.33） significantly increased the intracellular calcium concentration. Moreover, PRN synergized with As2O3 to increase the intracellular calcium concentration （1.34±0.72） compared with the findings in the As203 only group. The synergy of PRN and As2O3 significantly downregnlated phosphorylated Akt expression, as well as upregulated the levels of phosphorylated p38-MAPK, cleaved caspase-3 proteins, and caspase-3 mRNA. Conclusion： PRN synergized with AS2O3 tO increase intracellular calcium and inhibit cell survival. In addition, PRN synergized with As2O3 to repress phosphorylated Akt expression, as well as to upregulate phosphorylated p38-MAPK and cleaved caspase-3 protein levels, thereby contributing to cell apoptosis. Clinically, PRN combined with As2O3 is a potential adjuvant drug for the treatment of malignant tumors.

关 键 词：胶质瘤 P-AKT pp38-MAPK Cleaved Caspase-3凋亡 

分 类 号：R739.4[医药卫生—肿瘤]