二甲双胍对胃癌细胞糖代谢及其相关蛋白HIF-1α、PKM2表达的影响

Effects of metformin on glucose metabolism and expression of related proteins HIF-1α and PKM2 of gastric cancer cells

目的探讨二甲双胍对胃癌MGC-803细胞糖代谢及其相关蛋白缺氧诱导因子-1α(HIF-1α)、M2型丙酮酸激酶(PKM2)表达的影响。方法体外培养MGC-803细胞,将其分为阴性对照组和二甲双胍低、中、高剂量组,二甲双胍低、中、高剂量组分别予以1、5、10mmol/L二甲双胍,应用葡萄糖检测分析试剂盒检测二甲双胍对MGC-803细胞葡萄糖摄取的影响;应用乳酸检测试剂盒检测二甲双胍对MGC-803细胞乳酸生成量的影响;Westernblotting法检测MGC-803细胞中HIF-1α、PKM2蛋白表达;Real-timePCR法检测MGC-803细胞中HIF-1α、PKM2mRNA表达。结果与阴性对照组比较,二甲双胍低、中、高剂量组葡萄糖摄取量、乳酸生成量均降低(P均<0.05),且高剂量组下降最明显(P均<0.01)。与阴性对照组比较,二甲双胍低、中、高剂量组HIF-1α、PKM2蛋白及mRNA表达降低(P均<0.05),其中高剂量组降低最明显。结论二甲双胍可抑制胃癌细胞糖代谢,影响癌细胞生长的能量供应,抑制糖代谢相关蛋白HIF-1α、PKM2表达。

To investigate the effects of the different concentrations of metformin on the glucose metabolism and the expression of hypoxia-inducible factor-1α(HIF-1α)and M2 pyruvate kinase(PKM2)of gastric carcinoma MGC-803 cells.Methods MGC-803 cells were cultured in vitro and divided into the negative control group and low-,medium-,and high-dose groups.Metformin was given by concerntrations of 1,5,and 10 mmol/L in low-,medium-,and high-dose groups,respectively;the effect of metformin on glucose uptake in MGC-803 cells was detected by using glucose intake test kit.The effect of metformin on the production of lactic acid in MGC-803 cells was detected by means of lactic acid production kit.The expression of HIF-1αand PKM2 in MGC-803 cells was detected by Western blotting;the real-time PCR was used to detect the expression of HIF-1αand PKM2 mRNA in MGC-803 cells.Results Compared with the negative control group,the glucose intake and lactic acid production in sugar metabolism decreased in the low-,medium-and high-dose groups(both P<0.05),with the most significant decrease in the high-dose group(P<0.01);the protein and mRNA expression of HIF-1αand PKM2 in MGC-803 cells decreased in the low-,medium-,and high-dose groups(all P<0.05),with the most significant decrease in the high-dose group.Conclusion Metformin can inhibit the glucose metabolism of gastric cancer cells,affect the energy supply of cancer cell growth,and inhibit the expression of glycometabolic-related proteins HIF-1αand PKM2.