摘要
目的探讨去泛素化酶抑制剂Au(PPh3)PT对胃癌细胞的毒性作用及机制。方法MTS方法检测Au(PPh3)PT对胃癌细胞MGC⁃803和SGC⁃7901活力的影响;倒置荧光显微镜观察或流式细胞术检测Au(PPh3)PT诱导胃癌细胞凋亡情况;Western blot方法分析Au(PPh3)PT对凋亡相关蛋白及泛素化蛋白的影响。结果Au(PPh3)PT明显抑制胃癌细胞的活力,呈浓度和时间依赖性。1.0μmol/L和1.5μmol/L的Au(PPh3)PT作用24 h,胃癌细胞凋亡率显著增加(P<0.05)。Au(PPh3)PT引起凋亡相关蛋白caspase⁃3、caspase⁃8、caspase⁃9前体减少,活化带增多及PARP切割带出现,诱导胃癌细胞总的及K48⁃链接的泛素化蛋白聚集增多。结论Au(PPh3)PT明显抑制胃癌细胞的活力及诱导其凋亡,作用机制与caspase系统激活和蛋白酶体抑制有关。
Objective To investigate the effect of Deubiquitinase inhibitor Au(PPh3)PT on inducing gastric cancer cells cytotoxicity.Methods MTS assay was performed to test the cell viability of MGC⁃803 and SGC⁃7901 cancer cells.Apoptotic rates were assessed by microscopy and flow cytometry analysis using Annexin V/PI double staining.Apoptosis related proteins and ubiquitinated proteins were detected by western blot analysis.Results We found that Au(PPh3)PT significantly decreased the cell viability in a dose⁃and time⁃dependent man⁃ner.The cell apoptosis rate was obviously higher than that in control group after 24 h treatment with 1.0μmol/L and 1.5μmol/L of Au(PPh3)PT(P<0.05).It was confirmed that Au(PPh3)PT resulted in a reduction in precursor forms of caspase⁃3,⁃8,and⁃9 with a concomitant increase in the cleaved forms of caspase⁃3,caspase⁃8,caspase⁃9,and PARP.Also,we found that a striking increase in total and K48⁃linked ubiquitinated proteins.Conclusion The results show that Au(PPh3)PT induced cytotoxicity and cell death in gastric cancer cells.This effect may be mediated by activated caspase protein and accelerated proteasome inhibition.
作者
张培全
黄擎天
龙惠丹
李小芬
ZHANG Peiquan;HUANG Qingtian;LONG Huidan;LI Xiaofen(Guangdong Provincial Key Laboratory of Protein Modification and Degradation,Guangzhou Medical University,Guangzhou 511436,China)
出处
《实用医学杂志》
CAS
北大核心
2021年第1期25-29,共5页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号:81602427)
广东省自然科学基金博士启动项目(编号:2016A030310281)
广东省中医药局面上科研项目(编号:20192047)。
