3-溴丙酮酸抑制人肝癌细胞株糖酵解的研究  

Experimental study of 3 - bromopyruvate inhibiting the aerobic glycolysis of hepatocarcinoma cells

作　　者：吴昊[1] 甘蕾[1] 夏瑞 包闰 黄强[2] 庄志祥[1] Wu Hao;Gan Lei;Xia Rui;Bao Run;Huang Qiang;Zhuang Zhixiang(Department of Oncology,the Second Affiliated Hospital of Soochow University,Suzhou 215004,China（Wu H,Gan L,Xia R,Zhnang ZX;Department of Neurosurgery,the Second Affiliated Hospital of Soo-chow University,Suzhou 215004,China;Department of Gastroenterology,the Seventh People＇s Hospital of Suzhou,Suzhou 215151,China)

机构地区：[1]苏州大学附属第二医院肿瘤科,215004 [2]苏州大学附属第二医院神经外科,215004 [3]苏州市第七人民医院消化科,215151

出　　处：《中华实验外科杂志》2018年第9期1663-1666,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金预研项目（SDFEYGJ1608）

摘　　要：目的观察3-溴丙酮酸（3-BrPA）对人肝癌细胞株糖酵解的影响。方法不同浓度3-BrPA处理人肝癌细胞HepG2和人正常肝细胞QSG-7701后，细胞活性检测试剂盒（CCKl8）检测细胞存活率，流式细胞仪检测细胞凋亡率，实时荧光定量聚合酶链反应（FQ-PCR）和Westernblot检测HepG2和QSG-7701经过3-BrPA处理前后细胞内c-Myc、单羧酸转运蛋白1（MCTl）mRNA和蛋白表达，酶耦联法检测上述细胞葡萄糖吸收、乳酸分泌及细胞内三磷酸腺苷（ATP）含量。结果细胞存活和凋亡结果显示随3-BrPA浓度增加，细胞生长抑制率和凋亡率随之增高。不同浓度3-BrPA（25、50、75μmol／L）处理HepG2细胞24h后，凋亡率分别为（4．64±2．68）％、（14．57±8．41）％、（40．55±23．41）％，对照组0μmol／L下HepG2凋亡率为（5．62±3．24）％，75μmol／L组与对照组比较差异有统计学意义（P=0．025）。在mRNA和蛋白水平上，与QSG-7701比较，HepG2中c-Myc、MCTl均高表达（P=0．001、0．000），且当3-BrPA浓度为75μmol／L时表达均下降（P=0．041、0．000）。糖代谢结果显示当3-BrPA浓度为75μmol／L时HepG2的葡萄糖吸收、乳酸分泌及细胞内ATP含量均逐渐降低（P=0．035、0．000、0．045），而QSG-7701中无改变（P=0．898、0．841、o．112）。结论3-BrPA通过抑制c-Myc／MCTl表达抑制HepG2的糖酵解，从而抑制HepG2细胞生长、促进细胞凋亡。Objective To study the effect of 3 -bromopyruvate （3-BrPA） on the aerobic glycol- ysis pathway of hepatocarcinoma cell HepG2. Methods The human hepatocarcinoma cell HepG2 and the human normal liver cell QSG - 7701 were treated with different concentration of 3 - BrPA, the cell viability was detected by cell counting kit - 8 （ CCK-8 ） assay. Morphological changes of the two cells were observed under microscope. The cell apoptosis rate was detected by flow cytometry. Real - time quantitative poly- merase chain reaction （FQ -PCR） and Western blotting were used for detecting the expression of c -Myc and monocarboxylate transporter 1 （MCT1） in HepG2 and QSG -7701 at mRNA and protein levels. The glucose uptake, lactate secretion and intracellular content of adenosine triphosphate （ATP） of the cells were measured by enzyme - linked method. Results The cell survival rate and the cell apoptosis rate were significantly increased with the increase of 3 - BrPA concentration. Apoptosis rates of HepG2 treated with different concentrations of 3 - BrPA （25, 50, 75 μmol/L） were （4. 64 ± 2. 68 ） %, （ 14. 57 ±8. 41 ） % , （40. 55 ± 23.41 ） % respectively, the apoptosis rate of control group （0 μmol/L） was （5.62 ± 3.24 ） % , and the concentration of 75 ixmoL/L was significantly different from that of the control group （P = 0. 025 ）. Compared with QSG -7701, the expression of c -Myc and MCT1 in HepG2 were significantly higher than those in QSG -7701 at mRNA and protein levels （P = 0. 001,0. 000）, and decreased when the concentra- tion of 3 - BrPA was 75 μmol/L （P =0. 041, 0. 000）. The resuhs of glucose metabolism analysis showed that the glucose uptake, lactate secretion and intracellular ATP of HepG2 decreased gradually when the concentration of 3 - BrPA was 75 txmol/L （ P = 0. 035, 0. 000, 0. 045 ） , but no significant change in QSG -770 （P=0. 898, 0. 841, 0. 112）. Conclusion 3 - BrPA can inhibit the growth of HepG2 and promote the cell apoptosis by inhibiting th

关 键 词：3-溴丙酮酸 肝癌 c-Myc 单羧酸转运蛋白1 有氧糖酵解 

分 类 号：R735.7[医药卫生—肿瘤]