西达本胺对结肠癌细胞能量代谢的调节作用

Regulation of energy metabolism in colon cancer cells by chidamide

目的考察西达本胺对结肠癌细胞HCT-8和HT-29能量代谢调节的作用。方法西达本胺5,10和20μmol·L^(-1)分别处理HCT-8和HT-29细胞,普通光学显微镜观察细胞形态变化,MTT法检测细胞存活,荧光细胞活性试剂盒检测ATP含量,糖酵解压力试剂盒检测代谢变化,荧光定量PCR和Western蛋白印迹法分别检测糖酵解关键酶乳酸脱氢酶A(LDH-A)在m RNA水平和蛋白水平的表达。结果与对照组相比,西达本胺处理组HCT-8和HT-29细胞形态不规则,出现变形、皱缩和细胞碎片,增殖抑制率显著升高(P<0.05);西达本胺5和10μmol·L^(-1)处理16 h,HCT-8和HT-29细胞内ATP总含量均无差异,而20μmol·L^(-1)处理组ATP总含量显著降低(P<0.05)。西达本胺20μmol·L^(-1)处理HCT-8和HT-29细胞16 h,有氧呼吸水平均无差异,而糖酵解ATP产生速率分别降低30.7%和37.9%(P<0.05);LDH-A m RNA水平无差异,蛋白水平显著下调(P<0.01)。结论西达本胺具有抑制结肠癌细胞糖酵解能力的作用,可能与下调LDH-A有关。

OBJECTIVE To observe the regulation effect of chidamide on energy metabolism in HCT-8 and HT-29 cells. METHODS HCT-8 and HT-29 cells were treated with chidamide 5,10 and20 μmol·L^- 1. Morphological changes of these cells were observed under an ordinary optical microscope.Cell proliferation was detected by MTT. ATP production was determined by Cell Titer-Glo assay kit.Metabolic changes were tested by glycolytic stress kit. The m RNA level of lactate dehydrogenase A（LDH-A）was analyzed by real-time quantitative PCR,whereas the protein level of LDH-A was analyzed by Western blotting. RESULTS Compared with control group,cell morphology of HCT-8 and HT-29 cells in chidamide treated group was irregular,accompanied by deformation,shrinkage and cell debris,and the inhibitory rate of proliferation increased（P〈0.05）. There was no significant difference in ATP total content between chidamide 5 and 10 μmol·L^-116 h treatment groups,but in chidamide 20 μmol·L^-1treatment group it was decreased（P〈0.05）. Chidamide 20 μmol·L^-1had no effect on oxygen consumption rate, but glycolysis ATP generation rate was reduced by 30.7% and 37.9%（P〈0.05）,respectively.Chidamide 20 μmol·L^-1had no effect on LDH-A m RNA level,but it decreased the protein level of LDH-A（P〈0.01）. CONCLUSION Chidamide can abate the respiratory metabolic ability of HCT-8and HT-29 cells. The mechanism may be related to the down-regulation of LDH-A.