摘要
目的:探究顺铂耐药的非小细胞肺癌细胞糖代谢的特点及抑制有氧糖酵解对非小细胞肺癌细胞顺铂耐药细胞的影响及其机制。方法:通过梯度倍增浓度法建立非小细胞肺癌顺铂耐药细胞模型(A549/DDP);酶联免疫吸附试验(enzyme linked immunosorbent assay,Elisa)检测A549/DDP与A549细胞两组葡萄糖消耗量、三磷酸腺苷(adenosine triphosphate,ATP)释放量、乳酸生成及丙酮酸生成量的差异;CCK-8法(cell counting kit-8)检测2-脱氧葡萄糖(2-deoxyglucose,2-DG)及顺铂处理后A549/DDP与A549细胞增殖能力的变化;Elisa检测2-DG处理A549/DDP细胞与A549细胞葡萄糖消耗量、ATP释放量、乳酸生成及丙酮酸生成量的差异;Elisa检测2-DG处理A549/DDP细胞与A549细胞后对非神经元性烯醇化酶(recombinant enolase 1,ENO1)、葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate dehydrogenase,G6PD)、缺氧诱导因子-α(hypoxia inducible factor-α,HIF-α)表达的影响。结果:与A549细胞相比,顺铂耐药细胞株短时间内生长速度无明显差异。A549/DDP细胞消耗葡萄糖量增多、有氧糖酵解产生ATP增多、乳酸生成及丙酮酸释放量也增多,有氧糖酵解能力明显增强,差异具有统计学意义。2-DG处理后能够显著逆转A549/DDP细胞对顺铂的耐药性,增强对顺铂的敏感性,达到抑制A549/DDP细胞增殖的效果,差异具有统计学意义。2-DG能够抑制A549/DDP细胞的葡萄糖摄取,减少ATP的释放,减少乳酸生成及丙酮酸的产生,差异具有统计学意义。与A549细胞相比,A549/DDP细胞ENO1、G6PD、HIF-α表达水平升高,2-DG抑制有氧糖酵解后三者表达水平均降低,差异具有统计学意义。结论:2-DG抑制非小细胞肺癌细胞顺铂耐药有氧糖酵解,可在一定程度上逆转其对顺铂的耐药性。
Objective:To investigate the characteristics of glucose metabolism in cisplatin resistant non-small cell lung cancer cells,and the effects and mechanism of inhibiting aerobic glycolysis on cisplatin resistant non-small cell lung cancer cells.Methods:Cisplatin resistant cell model of non-small cell lung cancer(A549/DDP)was established by gradient doubling concentration method.Enzyme linked immunosorbent assay(Elisa)was used to detect the differences in glucose consumption,adenosine triphosphate(ATP)release,lactate production,and pyruvate production between A549/DDP and A549 groups.Cell counting kit-8(CCK-8)was used to detect the changes in the proliferation ability of A549/DDP and A549/A549 after treatment with 2-deoxyglucose(2-DG)and cisplatin.Elisa was performed to detect the differences in glucose consumption,ATP release,lactate production,and pyruvate production between A549/DDP and A549 treated with 2-DG.Elisa test was used to detect the expression of recombinant enolase 1(ENO1),glucose-6-phosphate dehydrogenase(G6PD),hypoxia inducible factor-α(HIF-α)after 2-DG treatment of A549/DDP and A549.Results:Compared with A549 cells,there was no significant difference in the growth rate of cisplatin resistant cell lines in a short period of time.The consumption of glucose,the production of ATP by aerobic fermentation,the production of lactate and the release of pyruvate by A549/DDP cells were increased,and the aerobic glycolysis ability was significantly enhanced,and the differences were significant.2-DG treatment can significantly reverse the resistance of A549/DDP cells to cisplatin,enhance their sensitivity to cisplatin,and achieve the effect of inhibiting A549/DDP cell proliferation,and the difference was statistically significant.2-DG can inhibit glucose uptake,reduce ATP release,and reduce lactate and pyruvate production in A549/DDP cells,and the difference was statistically significant.Compared with A549,the expression level of ENO1,G6PD,HIF-αwas increased in A549/DDP.After 2-DG treatment,the expression of the three can be reversed,and the difference was statistically significant.Conclusion:2-DG inhibits the aerobic hydrolysis of cisplatin resistance in non-small cell lung cancer cells,which can to some extent reverse their resistance to cisplatin.
作者
丁世凯
张正华
张秀莉
DING Shikai;ZHANG Zhenghua;ZHANG Xiuli(Department of Clinical Laboratory,Gansu Provincial Cancer Hospital,Gansu Lanzhou 730050,China)
出处
《现代肿瘤医学》
CAS
2024年第11期1973-1977,共5页
Journal of Modern Oncology
基金
甘肃省兰州市科技发展指导性计划项目(编号:2022-ZD-79)。
关键词
非小细胞肺癌
顺铂耐药
有氧糖酵解
2-脱氧葡萄糖
non-small cell lung cancer
cisplatin resistance
aerobic glycolysis
2-deoxyglucose
