CAJ | 학술논문

目的：探讨3-溴丙酮酸（3-BP）增强肝癌细胞对顺铂敏感性的作用及其可能机制。方法MTT法检测3-BP、顺铂对HepG2、SMMC7721细胞的增殖抑制作用。选择低于半数抑制浓度（IC50）的100μmol/L的3-BP和8μmol/L的顺铂单独或联合处理细胞，PI单染流式细胞术检测细胞凋亡，caspase 3活性检测试剂盒测定caspase-3的活性变化，ATP检测试剂盒测定细胞内ATP水平，Western blot检测XIAP、PARP蛋白的表达。结果3-BP在50~400μmol/L浓度范围内对HepG2、SMMC7721细胞具有明显的增殖抑制作用（P<0.01），作用48 h的IC50分别为238.9±13.9μmol/L、278.7±11.7μmol/L；顺铂在2~32μmol/L浓度范围内对HepG2、SMMC7721细胞具有明显的增殖抑制作用（P<0.01），作用48 h的IC50分别为16.4±0.9μmol/L、20.9±1.8μmol/L。100μmol/L 3-BP与8μmol/L顺铂联合作用于HepG2、SMMC7721细胞48 h的增殖抑制率分别为（60.6±2.2）%、（56.8±2.3）%，明显高于对照组和单独用药组（P<0.01）。100μmol/L 3-BP与8μmol/L顺铂联合作用于HepG2、SMMC7721细胞48 h的凋亡率分别为（51.1±4.3）%、（46.5±3.9）%，较单用3-BP、顺铂的凋亡率明显提高（P<0.01）。结论3-BP能增强肝癌细胞HepG2、SMMC7721对顺铂诱导的凋亡的敏感性，其机制可能是通过引起细胞内ATP缺乏、下调XIAP蛋白的表达以及增加caspase-3的活性。
목적：탐토3-추병동산（3-BP）증강간암세포대순박민감성적작용급기가능궤제。방법MTT법검측3-BP、순박대HepG2、SMMC7721세포적증식억제작용。선택저우반수억제농도（IC50）적100μmol/L적3-BP화8μmol/L적순박단독혹연합처리세포，PI단염류식세포술검측세포조망，caspase 3활성검측시제합측정caspase-3적활성변화，ATP검측시제합측정세포내ATP수평，Western blot검측XIAP、PARP단백적표체。결과3-BP재50~400μmol/L농도범위내대HepG2、SMMC7721세포구유명현적증식억제작용（P<0.01），작용48 h적IC50분별위238.9±13.9μmol/L、278.7±11.7μmol/L；순박재2~32μmol/L농도범위내대HepG2、SMMC7721세포구유명현적증식억제작용（P<0.01），작용48 h적IC50분별위16.4±0.9μmol/L、20.9±1.8μmol/L。100μmol/L 3-BP여8μmol/L순박연합작용우HepG2、SMMC7721세포48 h적증식억제솔분별위（60.6±2.2）%、（56.8±2.3）%，명현고우대조조화단독용약조（P<0.01）。100μmol/L 3-BP여8μmol/L순박연합작용우HepG2、SMMC7721세포48 h적조망솔분별위（51.1±4.3）%、（46.5±3.9）%，교단용3-BP、순박적조망솔명현제고（P<0.01）。결론3-BP능증강간암세포HepG2、SMMC7721대순박유도적조망적민감성，기궤제가능시통과인기세포내ATP결핍、하조XIAP단백적표체이급증가caspase-3적활성。
Objective To investigate the effect of 3-bromopyruvate (3-BP) in sensitizing hepatocellular carcinoma cells to cisplatin-induced apoptosis and its possible mechanism. Methods 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 100μmol/L 3-BP with or without 8μmol/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. Results 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 2-32μmol/L, with IC50 values of 16.4±0.9μmol/L and 20.9±1.8μmol/L after a 48-h treatment, respectively. Treatment with 100μmol/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. Conclusion 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.