山竹醇对人口腔鳞癌细胞糖酵解代谢通路的影响  

Effects of Garcinol on glycolysis in oral squamous cell carcinoma cells

作　　者：张桂莲[1] 宿颖[1] 张辛燕[1] 

机构地区：[1]首都医科大学口腔医学院口腔医学研究所,北京100050

出　　处：《北京口腔医学》2017年第2期66-70,共5页Beijing Journal of Stomatology

基　　金：国家自然科学基金(81272982);北京市自然科学基金(7162073)

摘　　要：目的观察山竹醇(Garcinol)对人口腔鳞癌细胞系CAL27增殖和克隆形成能力的影响,以及对CAL27细胞糖酵解代谢的影响。方法培养人口腔鳞癌细胞系CAL27,不同浓度的Garcinol处理CAL27细胞,MTT法检测其对细胞增殖能力的影响。克隆形成实验检测对细胞克隆形成能力的影响。RT-PCR检测Garcinol对细胞内糖酵解通路关键酶表达的影响,以及对糖酵解通路的重要基因表达的影响。并检测细胞上清液中葡萄糖、乳酸含量的变化来评估Garcinol对CAL27细胞内糖酵解水平的影响。通过酶活性实验检测细胞内糖酵解关键酶(HK、PK、LDH)的酶活性变化。结果 Garcinol能显著抑制CAL27细胞的增殖,呈浓度和时间依赖性(P<0.01);能明显抑制CAL27细胞的克隆形成能力(P<0.01)。同时,Garcinol能促进糖酵解关键酶己糖激酶(HK),丙酮酸激酶(PK),乳酸脱氢酶(LDH)的基因表达,增强糖酵解关键酶(HK、PK、LDH)的酶活性(P<0.05)。Garcinol也能增加AKT和m TOR的基因表达水平(P<0.05)。Garcinol还能促进CAL27细胞的葡萄糖吸收和乳酸分泌。结论 Garcinol能够抑制人口腔鳞癌细胞系CAL27的增殖和克隆形成能力,同时也增加了细胞内的糖酵解水平。Objective To investigate the roles of garcinol in proliferation and colony forming ability, and explore the effects of garcinol on glycolysis in human oral squamous cell carcinoma （OSCC） cell line CAL27. Methods OSCC cell line CAL27 was cultured in DMED high glucose medium with 10% FBS and treated with low to high concentrations of garcinol. Cell proliferation was evaluated by MTT assay. Colony formation was detected by colony formation assay. The gene expression of key glycolysis enzymes, hexokinase （HK）, pyruvate kinase （PK）, lactate dehydrogenase （LDH） , as well as genes related to glycolysis, protein kinase B （AKT）, the mammalian target of rapamycin （roTOR） were measured by RT-PCR. The content of glucose and lactic acid in the cell supernatant was measured by spectrophotometric method. Enzymatic activity of glycolysis key enzymes （ HK, PK, LDH） was examined by enzyme activity assay. Results Garcinol could inhibit CAL27 cell proliferation and colony formation in a dose-dependent and time-dependent manner. Garcinol increased the gene expression of key glycolytic enzymes （ HK, PK, LDH） and AKT and roTOR. Garcinol also increased the level of intracellular glycolysis and the activity of key glycolysis enzymes. Conclusion As a natural agent, gareinol inhibited OSCC cells proliferation and colony formation and showed glycolysis promoting potential.

关 键 词：山竹醇 口腔鳞癌 糖酵解 

分 类 号：R739.8[医药卫生—肿瘤]