目的 探讨维生素 C 与热处理对肺癌 A549 细胞的作用及相关机制。 方法 应用不同浓度的维生素 C(终浓度 0、1、 2、4、8 mmol / L)与不同温度(37 ℃和 41 ℃ )作用于 A549 细胞,采用 CCK8 法检测细胞活性,FRASC 维生素 C 检测分析试剂盒 Ⅱ检测细胞内总维生素 C 的浓度;然后分为对照组(37 ℃ ,0 mmol / L 维生素 C)、维生素 C 组(37 ℃ ,8 mmol / L 维生素 C)、热 处理组(41 ℃ ,0 mmol / L 维生素 C)、联合组(41 ℃ ,8 mmol / L 维生素 C)干预 24 h,荧光标记 2-脱氧葡萄糖(2-NBDG)检测 A549 细胞葡萄糖摄取量,蛋白质印迹检测 GLUT1、GLUT3、PI3K、AKT、p-PI3K、p-AKT、HIF-1α 蛋白表达水平,检测结果均应 用方差分析及 LST-t 检验进行统计分析。 结果 A549 细胞活力随维生素 C 浓度增加而下降(P<0. 05),41 ℃ 热处理的细胞在 维生素 C 干预后的细胞活力均低于 37 ℃ (P<0. 05);细胞内总维生素 C 水平随干预浓度增加而升高,热处理的细胞在 2、4、 8 mmol / L 维生素 C 干预后,细胞内维生素 C 水平均高于 37 ℃ (P<0. 05);分组干预后,与对照组相比,维生素 C 组、热处理组 及联合组的细胞内葡萄糖摄取量均降低(P<0. 05),且热处理组低于维生素 C 组(P<0. 05),而联合组又低于维生素 C 组和热 处理组(P<0. 05);三个干预组的 GLUT1、PI3K、p-PI3K、p-AKT 蛋白表达水平均低于对照组(P<0. 05),维生素 C 组的 GLUT3、 HIF-1α,及联合组的 GLUT3、HIF-1α、AKT 均低于对照组(P<0. 05);联合组的 GLUT1、PI3K、p-PI3K 蛋白表达水平低于维生 素 C 组(P<0. 05),联合组的 GLUT3、AKT、p-PI3K、HIF-1α 均低于维生素 C 组和热处理组(P<0. 05)。 结论 维生素 C 联合热 处理显著地抑制 A549 细胞 PI3K/ AKT/ HIF-1α 通路,下调 GLUT1 和 GLUT3 的表达,减少细胞内的葡萄糖摄取,抑制 A549 细 胞增殖。

Objective To investigate the effects of vitamin C and heat treatment on lung cancer A549 cells and the related mechanisms. Method Different concentrations of vitamin C The final concentration is 0 1 2 4 8 mmol / L 0 1 2 4 8 mmol / L and different temperatures 37 ℃ and 41 ℃ were applied to A549 cells. CCK8 assay was used to detect the cell activity and FRASC Vitamin C assay Kit II was used to detect the concentration of vitamin C in the cells. Then A549 cells were divided into control group 37 ℃ 0 mmol / L vitamin C vitamin C group 37 ℃ 8 mmol / L vitamin C heat treatment group 41 ℃ 0 mmol / L vitamin C and combined group 41 ℃ 8 mmol / L vitamin C after 24 hours of intervention 2-NBDG was used to detect glucose uptake of A549 cells. The expression levels of GLUT1 GLUT3 PI3K AKT p-PI3K p-AKT and HIF-1α were detected by Western Blot assay. The results were statistically analyzed by ANOVA and LST-t test. Result The cell viability of A549 decreased with the increase of vitamin C intervention concentration P<0. 05 and the cell viability of 41 ℃ heat treated cells was lower than 37 ℃ after vitamin C intervention. P < 0. 05 . The intracellular vitamin C level increased with the increase of intervention concentration and the intracellular vitamin C level of 41 ℃ heat treated cells was lower than 37 ℃ after 2 4 and 8 mmol / L vitamin C intervention. P< 0. 05 . After the intervention compared to the control group the intracellular glucose intake of vitamin C group heat treatment group and combined group was decreased P<0. 05 and the heat treatment group was lower than the vitamin C group P<0. 05 and the combined group was lower than the vitamin C group and heat treatment group P < 0. 05 . Western Blot results showed that the expression levels of GLUT1 PI3K P-PI3K p-AKT in the three intervention groups and GLUT3 HIF-1α in the vitamin C group,and GLUT3 HIF-1α and AKT in the combined group were lower than those in control group P<0. 05 . The protein expression levels of GLUT1 PI3K p-PI3K in the combined group were lower than those in the vitamin C group P< 0. 05 and GLUT3 AKT P-PI3K and HIF-1α in the combined group were lower than those in the vitamin C group and the heat treatment group P<0. 05 . Conclusion Compared with vitamin C or 41 ℃ heat treatment vitamin C combined heat treatment significantly inhibited the PI3K/ AKT/ HIF-1α pathway of A549 cells down-regulated GLUT1 and GLUT3 reduced intracellular glucose uptake and inhibited A549 cell proliferations.