Zhao Surong, Zhang Yuanyuan, Wu Chengzhu, Li Hongmei, Jiang Chenchen, Jiang Zhiwen, Liu Hao
Faculty of Pharmacy, Bengbu Medical College/Anhui Engineering Technology Research Center of Biochemical Pharmaceuticals, Bengbu 233030, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2014 Jan;34(1):25-30.
To investigate the effect of 3-bromopyruvate (3-BP) in sensitizing hepatocellular carcinoma cells to cisplatin-induced apoptosis and its possible mechanism.
The growth inhibition of HepG2 and SMMC7721 cells following exposures to different concentrations of 3-BP and cisplatin was measured by MTT assay. The apoptosis of cells treated with 100 µmol/L 3-BP with or without 8 µmol/L cisplatin was assessed using flow cytometry with PI staining, and the activity of caspase-3 and intracellular ATP level were detected using commercial detection kits; the expression of XIAP and PARP was analyzed using Western blotting.
3-BP produced obvious inhibitory effects on HepG2 and SMMC7721 cells at the concentrations of 50-400 µmol/L with IC50 values of 238.9∓13.9 µmol/L and 278.7∓11.7 µmol/L for a 48-h treatment, respectively. Cisplatin also inhibited the growth of HepG2 and SMMC7721 cells at the concentrations of 2-32 µmol/L, with IC50 values of 16.4∓0.9 µmol/L and 20.9∓1.8 µmol/L after a 48-h treatment, respectively. Treatment with 100 µmol/L 3-BP combined with 8 µmol/L cisplatin for 48 h resulted in a growth inhibition rate of (60.6∓2.2)% in HepG2 cells and (56.8∓2.3)% in SMMC7721 cells, which were significantly higher than those in cells treated with 3-BP or cisplatin alone. The combined treatment for 48 h induced an apoptotic rate of (51.1∓4.3)% in HepG2 cells and (46.5∓3.9)% in SMMC7721 cells, which were also markedly higher than those in cells with 3-BP or cisplatin treatment alone.
3-BP can sensitize HepG2 and SMMC7721 cells to cisplatin-induced apoptosis possibly by causing intracellular ATP deficiency, down-regulating XIAP, and increasing caspase-3 activity.
探讨3-溴丙酮酸（3-BP）对肝癌细胞顺铂诱导凋亡的增敏作用及其可能机制。
采用MTT法检测不同浓度的3-BP和顺铂作用后HepG2和SMMC7721细胞的生长抑制情况。运用PI染色通过流式细胞术评估100 μmol/L 3-BP单独或联合8 μmol/L顺铂处理后细胞的凋亡情况，使用商业检测试剂盒检测caspase-3活性和细胞内ATP水平；采用蛋白质印迹法分析XIAP和PARP的表达。
50 - 400 μmol/L浓度的3-BP对HepG2和SMMC7721细胞产生明显抑制作用，48小时处理的IC50值分别为238.9±13.9 μmol/L和278.7±11.7 μmol/L。2 - 32 μmol/L浓度的顺铂也抑制HepG2和SMMC7721细胞生长，48小时处理后的IC50值分别为16.4±0.9 μmol/L和20.9±1.8 μmol/L。100 μmol/L 3-BP联合8 μmol/L顺铂处理48小时后，HepG2细胞的生长抑制率为（60.6±2.2）%，SMMC7721细胞为（56.8±2.3）%，显著高于单独使用3-BP或顺铂处理的细胞。联合处理48小时诱导HepG2细胞凋亡率为（51.1±4.3）%，SMMC7721细胞为（46.5±3.9）%，也明显高于单独使用3-BP或顺铂处理的细胞。
3-BP可能通过导致细胞内ATP缺乏、下调XIAP以及增加caspase-3活性，使HepG2和SMMC7721细胞对顺铂诱导的凋亡敏感。
