SHIP2通过上调Bim表达增强胃癌细胞BGC-823对紫杉醇的敏感性  

SHIP2 sensitizes gastric cancer cells BGC-823 to paclitaxel through upregulation of Bim

作　　者：葛燕梅[1] 叶艳[1] 张林杰[1] 

机构地区：[1]安徽医科大学基础医学院免疫学教研室,安徽合肥230032

出　　处：《中国药理学通报》2014年第5期701-706,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81301738);安徽省高等学校省级优秀青年人才基金资助项目(No 2012SQRL069);安徽省教育厅自然科学基金项目(KJ2011A167);安徽省自然科学基金项目(1408085QH150)

摘　　要：目的探讨SHIP2(SH2 domain containing inositol 5-phosphatase 2)基因转染人胃癌细胞BGC-823后对紫杉醇敏感性的影响。方法流式细胞仪PI单染法检测细胞凋亡率。应用免疫蛋白印迹法(Western blot)检测各组细胞蛋白表达情况。实时荧光定量PCR(real-time quantitative PCR,qRT-PCR)分析mRNA的变化水平。将pCMV6-SHIP2质粒在脂质体介导下转染人胃癌BGC-823细胞,同时以转染空载体和未转染细胞作为对照组。将GV112-Puromycin空载体和GV112-Puromycin-Bim(Bcl-2 interacting mediator of cell death)慢病毒颗粒分别感染BGC-823细胞,建立稳定细胞株,再将pCMV6-SHIP2质粒分别转染空载体和稳定干扰Bim表达细胞株。结果与SGC-7901细胞相比,BGC-823细胞对紫杉醇诱导的凋亡相对不敏感,0.3μmol·L-1紫杉醇诱导BGC-823 48 h后细胞凋亡率仅为(25.6±1.6)%,在紫杉醇作用不同时间点Bim蛋白和mRNA表达均无明显变化;与对照组相比,转染SHIP2基因能明显增加Bim的表达,紫杉醇作用48 h后,细胞凋亡率上升到(50.8±0.9)%;GV112-Puromycin-Bim慢病毒颗粒感染的BGC-823细胞,Bim的表达明显被抑制,在GV112-Puromycin-Bim慢病毒颗粒感染的稳定BGC-823细胞中转入pCMV6-SHIP2,紫杉醇作用48 h后,凋亡率为(27.6±1.6)%。结论转染外源性SHIP2基因能有效提高BGC-823细胞内Bim的表达,诱导BGC-823细胞凋亡,明显增加BGC-823细胞对紫杉醇的敏感性。Aim To study the sensitivity of human gastric cancer BGC -823 cells to paclitaxel after transfection of SHIP2 （The SH2 domain containing inositol 5-phosphatase 2） cDNA. Methods Apoptotic cells were determined by the propidium iodide method using flow cytometry. The levels of protein and mRNA expression were measured by Western blot analysis and qRT-PCR, respectively, pCMV6-SHIP2 plasmid and empty vector were transiently transfected into BGC-823 cells, respectively. Stable cell lines were established after infecting BGC-823 cell with GV112-Puromycin and GV112-Puromycin-Bim（ Bcl-2 interacting mediator of cell death） lentivirus particles, pCMV6-SHIP2 plasmid was transiently transfected into the stable cell lines. Results BGC-823 cells were relatively insensitive to paclitaxel compared with SGC-7901 cells. The ap0pt0tic rate was only （25.6 ± 1.6） % after the treatment with 0.3 umol . L-1 paclitaxel for 48h in BGC- 823 cells. The expression levels of Bim protein and mRNA in BGC-823 cells treated with paclitaxel at different time points were not significantly changed. The expression of Bim protein was increased after transfection of pCMV6-SHIP2 plasmid, and the apoptotie rate was up to （50. 8 ± 0.9 ）% in BGC-823 cells treated with paclitaxel for 48h. The expression of Bim protein was significantly inhibited after infecting with GV112- Puromycin-Bim lentivirus particles. The apoptotic rate of infected BGC-823 cells was only （27.6 ± 1.6）% after treatment upon paelitaxel for 48h. Conclusion Overexpression of exogenous SHIP2 can increase the expression of Bim, induce apoptosis and enhance sensitivity of BGC-823 cells to paclitaxel.

关 键 词：SHIP2 胃癌 BIM 紫杉醇 细胞凋亡 AKT 

分 类 号：R282.71[医药卫生—中药学] R329.25[医药卫生—中医学]