Abstract:

Objective To investigate whether tumor suppressor N-Myc downstream regulated gene 2（NDRG2）participates in regulating glucose metabolism in colon tumor cells. Methods NDRG2 was stably transfected in colon cancer cells HCT116. Lactate production was measured by lactate assay kits, and glucose consumption was measured by glucose assay kit. Besides, western blot was used to examine the expression of PKM、LDHA、HK, as the key glycolytic enzymes. Scratch test was used to test tumor cells proliferation. Results HCT116 which transfected with NDRG2 produced less lactate compared with control（38.2 ± 3.4 mmol/mL vs 62.1 ± 4.8 mmol/mL, P<0.05）, and remaining glucose was more than the control（137 ± 3.6 mmol/mL vs 88.2 ± 2.2 mmol/ml, P<0.05）. Western blot showed the expression of PKM、LDHA、HK were less than that in control. Scratch test showed less proliferation of colon tumor cells when above protein were inhibited. Conclusion NDRG2 functions to inhibits glycolysis of colon cancer cells as tumor suppressor gene by repressing the glycolytic enzymes expression, and then inhibits tumor cells proliferation.

Key words: NDRG2, colon cancer, glycolysis

摘要：

目的 探讨肿瘤抑制因子N-myc下游调节基因2（NDRG2）于结肠癌细胞葡萄糖代谢调节，并通过调控糖酵解抑制结肠癌细胞增殖的作用。方法 稳定转染NDRG2于结肠癌细胞HCT116，乳酸检测试剂盒检测细胞（1×106）乳酸代谢的水平；葡萄糖试剂盒检测通过细胞（1×106）消耗培养基葡萄糖的水平；Westernblot检测HCT116细胞（1×106）糖酵解中丙酮酸激酶（PKM）、乳酸脱氢酶（LDHA）、己糖激酶（HK）表达水平，划痕实验检测肿瘤细胞增殖能力。结果 转染NDRG2后，HCT116细胞生成乳酸量少于对照组38.2±3.4mmol/mL比62.1±4.8mmol/mL，P<0.05），培养基剩余葡萄糖量则多于对照组（137±3.6mmol/mL比88.2±2.2mmol/mL，P<0.05）。Westernblot检测NDRG2转染的HCT116细胞显示PKM、LDHA、HK表达显著低于对照组HCT116细胞。划痕实验证实，抑制结肠癌细胞糖酵解后能够抑制其增殖作用。结论 NDRG2作为肿瘤抑制基因，通过抑制糖酵解关键酶生成，抑制肿瘤细胞糖酵解，进而抑制肿瘤细胞增殖。

关键词: 糖酵解, 结肠癌 , NDRG2