文献类型：期刊文献

中文题名：甘草黄酮A对低氧条件下胃癌细胞增殖和糖酵解的影响

英文题名：Effect of Licoflavone A on Proliferation and Glycolysis of Gastric Cancer Cells Under Hypoxic Conditions

作者：董焕成[1];苏韫[1];龚红霞[1];曹旺杰[1];袁敏捷[1];刘永琦[1];黄勇[1]

第一作者：董焕成

机构：[1]甘肃中医药大学基础医学院,甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,敦煌医学与转化教育部重点实验室,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）|甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2024

卷号：30

期号：13

起止页码：120

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;Scopus;北大核心:【北大核心2023】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金地区科学基金项目(82360928);兰州市科技计划项目(2023-2-85);甘肃省教育厅青年博士支持项目(2023QB-087);甘肃中医药大学研究生创新创业基金项目(2023CXZX-748);甘肃省“双一流”科研重点项目(GSSYLXM-05)。

语种：中文

中文关键词：甘草黄酮A;胃癌;低氧;糖酵解;增殖

外文关键词：licoflavone A;gastric cancer;hypoxia;glycolysis;proliferation

摘要：目的:探讨低氧条件下甘草黄酮A对胃癌细胞增殖和糖酵解的影响。方法:筛选人胃癌AGS细胞予以5%O2低氧环境培养48 h,分为5组:常氧组、低氧组、甘草黄酮A高、中、低浓度组(100、50、25μmol·L^(-1))。细胞增殖与活性检测(CCK-8)法和克隆形成实验检测甘草黄酮A对低氧条件下AGS细胞增殖的影响;划痕实验检测细胞迁移情况,蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链式反应(Real-time PCR)检测AGS细胞中低氧诱导因子-1α(HIF-1α)、葡萄糖转运蛋白1(GLUT1)、乳酸脱氢酶A(LDHA)、丙酮酸激酶M2(PKM2)、己糖激酶Ⅱ(HK2)的蛋白及mRNA表达水平。试剂盒检测葡萄糖摄取和HK的活性的表达情况。结果:CCK-8法实验结果表明,与低氧组比较,24 h时,甘草黄酮A高、中浓度组AGS细胞的增殖率显著降低(P<0.01),48 h时,甘草黄酮A高、中、低浓度组AGS细胞的增殖率显著降低(P<0.01)。克隆形成实验结果表明,与常氧组比较,低氧组AGS细胞克隆形成数显著升高(P<0.01);与低氧组比较,甘草黄铜A高、中、低浓度组AGS细胞的克隆形成数显著降低(P<0.01)。划痕实验表明,与常氧组比较,低氧组的AGS细胞迁移的能力升高(P<0.01);与低氧组比较,甘草黄酮A高、中、低浓度组的AGS细胞各个时间段迁移细胞的能力显著降低(P<0.01)。Real-time PCR表明,与常氧组比较,低氧组GLUT1、LDHA、PKM2和HK2 mRNA明显升高(P<0.05,P<0.01);与低氧组比较,甘草黄酮A高浓度组GLUT1、LDHA、PKM2、HK2 mRNA,甘草黄酮A中浓度组GLUT1、LDHA、HK2 mRNA、甘草黄酮A低浓度组HK2 mRNA明显降低(P<0.05,P<0.01)。Western blot表明,与常氧组比较,低氧组HIF-1α、GLUT1、LDHA、PKM2和HK2蛋白的表达明显升高(P<0.05,P<0.01);与低氧组比较,甘草黄酮A高浓度组的HIF-1α、GLUT1、LDHA、PKM2和HK2蛋白的表达明显降低(P<0.05,P<0.01),甘草黄酮A中、低浓度的HK2蛋白的表达明显降低(P<0.05,P<0.01)。试剂盒检测葡萄糖摄取及HK活性结果表明,与常氧组比较,低氧组葡萄糖的摄取和HK的含量表达升高(P<0.01),与低氧组比较,甘草黄酮A高浓度组葡萄糖的摄取和HK的含量表达及甘草黄酮A中浓度组HK的含量表达降低(P<0.01)。结论:甘草黄酮A可以抑制低氧条件下AGS细胞的增殖,其作用机制可能与抑制胃癌的糖酵解有关。
Objective:To investigate the effects of licoflavone A on the proliferation and glycolysis of gastric cancer cells in the hypoxic environment.Method:Human gastric cancer AGS cells were classified into five groups:Normoxia,hypoxia,and low-,medium-,and high-dose(25,50,100μmol·L^(-1),respectively)licoflavone A.The cells in other groups except the normoxia group were cultured in the environment with 5%O2 for 48 h.The cell counting kit-8(CCK-8)and colony formation assay were employed to examine the proliferation of AGS cells.Cell migration was detected by the scratch assay.The protein and mRNA levels of hypoxia-inducible factor 1-alpha(HIF-1α),glucose transporter 1(GLUT1),lactate dehydrogenase A(LDHA),pyruvate kinase M2(PKM2),and hexokinaseⅡ(HK2)in AGS cells were measured by Western blotting and real-time quantitative polymerase chain reaction(Real-time PCR),respectively.The corresponding kits were used to determine glucose uptake and HK activity.Result:The CCK-8 results showed that compared with the hypoxia group,the high-and medium-dose licoflavone A groups showed decreased proliferation rate of AGS cells at the time point of 24 h(P<0.01)and all the licoflavone A groups demonstrated decreased proliferation rate at the time point of 48 h(P<0.01).Compared with the normoxia group,the hypoxia group showed increased number of clone formation of AGS cells(P<0.01),which was decreased after the treatment with licoflavone A at high,medium,and low doses(P<0.01).Compared with the normoxia group,the hypoxia group showed increased migration of AGS cells(P<0.01),which was attenuated by the high,medium,and low doses of licoflavone A(P<0.01).Compared with the normoxia group,the hypoxia group showed up-regulated mRNA levels of GLUT1,LDHA,PKM2,and HK2(P<0.05,P<0.01).Compared with those in the hypoxia group,the mRNA levels of GLUT1,LDHA,PKM2,and HK2 in the high-dose licoflavone A group,GLUT1,LDHA,and HK2 in the medium-dose licoflavone A group,and HK2 in the low-dose licoflavone A group were down-regulated(P<0.05,P<0.01).The protein levels of HIF-1α,GLUT1,LDHA,PKM2,and HK2 in the hypoxia group were higher than those in the normoxia group(P<0.05,P<0.01).Compared with those in the hypoxia group,the protein levels of HIF-1α,GLUT1,LDHA,PKM2,and HK2 in the high-dose licoflavone A group and HK2 in the medium-and low-dose licoflavone A groups were down-regulated(P<0.05,P<0.01).The glucose uptake and HK activity were elevated in the hypoxia group compared with those in the normoxia group(P<0.01).Compared with the hypoxia group,high-dose licoflavone A decreased the glucose uptake and HK activity,and medium-dose licoflavone A decreased the HK activity(P<0.01).Conclusion:Licoflavone A inhibits the proliferation of AGS cells under hypoxic conditions by regulating glycolysis in gastric cancer.