山东医药2019，Vol.59Issue(2)：1-5,5.DOI:10.3969/j.issn.1002-266X.2019.02.001
2-D-脱氧葡萄糖对二甲双胍抑制人结肠癌细胞作用的影响及机制
2-DG enhances anti-tumor effect of metformin on human colon cancer cells
摘要
Abstract
Objective To investigate the role of hexokinase inhibitor 2-deoxy-D-glucose ( 2-DG) in enhancing antitumor effect of metformin on human colon cancer cells and its mechanism. Methods HT-29 human colon cancer cells were treated with different dosages of 2-DG and metformin alone or jointly. The cell viability and proliferation rate were determined by trypan blue staining and MTT assay, respectively. The apoptotic rate was recorded by a double staining with Annexin V/PI followed by FACS analysis. Western blotting was used to detect the protein levels of several key components involved in AKT/mTOR signaling pathway and autophagy, including AKT, phosphorylated AKT ( p-AKT) , ribosomal p70S6 kinase ( p70S6K) , phosphorylated ribosomal p70S6 kinase ( p-p70S6K) , and p62. Results After combined treatment of 2-DG ( 1, 5, and 10 mmol/L) and metformin ( 5 and 10 mmol/L) for 24 h, the cell mortality rates of HT-29 cells were much higher than those of cells receiving single drug treatment ( all P < 0. 05) , with the highest mortality rate at 10mmol/L of both drugs ( P < 0. 01) . The combined treatment of 10 mmol/L 2-DG and various amount of metformin ( 0, 1, 5, 10, 15, and 20 mmol/L) for 48 h inhibited the cell proliferation as compared with the corresponding single drug treatment when the metformin was ≥ 5 mmol/L ( all P < 0. 05) and the cell viability was the lowest at 10 mmol/L metformin ( P < 0. 01) , however, when the concentration was 20 mmmol/L, the survival rate of HT-29 cells did not continue to decrease significantly. HT-29 cells were treated with different dosages of 2-DG ( 5 and 10 mmol/L) and 10 mmol/L of metformin for 24 h, but only under the combined treatment of 10 mmol/L 2-DG and metformin, the apoptosis rate of HT-29 cells was higher than that under the single drug treatment ( all P < 0. 05) , as well as the expression of p-AKT, p-p70S6 K and p62 proteins. Conclusion 2-DG enhances the efficacy of metformin in inhibiting the viability and proliferation of HT-29 cells and induces the apoptosis, with maximum effect at the dosage of 10 mmol/L, and the mechanism may be related to their synergistic inhibition of AKT/mTOR pathway and autophagy in tumor cells.关键词
结直肠癌/2-D-脱氧葡萄糖/二甲双胍/AKT/mTOR信号通路/细胞自噬/p62蛋白Key words
colorectal carcinoma/2-deoxy-D-glucose/metformin/AKT/mTOR signaling pathway/autophagy/p62 protein分类
医药卫生引用本文复制引用
谢敏,李红霞,顾馨仪,郝舒捷,王仁军,刘庆平,秦建中,张帆..2-D-脱氧葡萄糖对二甲双胍抑制人结肠癌细胞作用的影响及机制[J].山东医药,2019,59(2):1-5,5.基金项目
辽宁省自然科学基金资助项目(20170540033). （20170540033）
