金复康口服液对吉非替尼耐药肺腺癌H1975细胞有氧糖酵解的影响  

Effect of Jinfukang oral liquid on aerobic glycolysis of gefitinib resistant lung adenocarcinoma H1975 cells

作　　者：张轶 贾方 王秋月 崔文静 王菊勇[1] ZHANG Yi;JIA Fang;WANG Qiuyue;CUI Wenjing;WANG Juyong(Longhua Hosptial,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院,上海200032

出　　处：《现代中西医结合杂志》2024年第2期155-161,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：十三五“重大新药创制”科技重大专项(2017ZX09304001);龙华医院科技创新项目(CX201838)。

摘　　要：目的 探讨金复康口服液联合吉非替尼对肺腺癌H1975细胞有氧糖酵解的影响。方法 首先采用CCK-8法检测不同浓度金复康口服液和吉非替尼对H1975细胞增殖的影响;然后实验分为空白组、吉非替尼组、金复康口服液组和联合用药组,采用2-NBDG荧光探针和乳酸测试盒分别检测各组细胞葡萄糖摄取能力及乳酸生成量,酶活性试剂盒检测细胞中糖酵解关键限速酶[己糖激酶(HK)、磷酸果糖激酶(PFK)、丙酮酸激酶(PK)]的活性,Western blot法检测细胞中己糖激酶2(HK2)、P型磷酸果糖激酶(PFKP)、M2型丙酮酸激酶(PKM2)蛋白表达情况,JC-1、DCFH-DA荧光探针分别检测细胞线粒体膜电位和活性氧(ROS)水平。结果 金复康口服液对H1975细胞的生长抑制作用呈时间和剂量依赖性增强,差异均有统计学意义(P均<0.05);与不同浓度吉非替尼单独作用48 h比较,相应浓度吉非替尼联合金复康口服液作用后的H1975细胞增殖抑制率均更高(P均<0.05)。各给药组细胞葡萄糖相对摄取量与空白组比较差异均无统计学意义(P均>0.05);各给药组细胞乳酸生成量均明显低于空白组(P均<0.05),且联合用药组明显低于吉非替尼组和金复康口服液组(P均<0.05);吉非替尼组PFK、PK活性,金复康口服液组HK、PFK活性,联合用药组HK、PFK、PK活性均明显低于空白组(P均<0.05),且金复康口服液组与联合用药组HK、PFK活性均明显低于吉非替尼组(P均<0.05)。金复康口服液组和联合用药组HK2、PFKP、PKM2蛋白相对表达量及吉非替尼组PFKP蛋白相对表达量均明显低于空白组(P均<0.05),且金复康口服液组和联合用药组HK2、PFKP、PKM2蛋白相对表达量均明显低于吉非替尼组(P均<0.05)。与空白组比较,各给药组细胞线粒体膜电位明显下降,ROS明显增加,且联合用药组细胞线粒体膜电位下降和ROS增加更明显。结论 金复康口服液能通过抑制有氧糖酵解途径,降低细胞线粒体膜Objective It is to investigate the effect of Jinfukang oral liquid(JFK) combined with gefitinib on aerobic glycolysis of lung adenocarcinoma H1975 cells.Methods Firstly,the effects of different concentrations of JFK and gefitinib on the proliferation of H1975 cells were detected by cell counting kit(CCK-8).Then the experiment was divided into blank group,gefitinib group,JFK group and combination group,the glucose uptake and lactate production of the cells in each group were detected by 2-NBDG and lactate test kit,the activities of key rate-limiting enzymes of glycolysis [hexokinase(HK),phosphofructokinase(PFK),pyruvate kinase(PK)] in cells were detected by enzyme activity kit,the protein expressions of hexokinase 2(HK2),P-type phosphofructokinase(PFKP),and M2-type pyruvate kinase(PKM2) in cells were detected by Western blot,and the cellular mitochondrial membrane potential and the level of reactive oxygen species(ROS) were detected by JC-1,DCFH-DA JC-1,DCFH-DA fluorescent probes.Results The growth inhibitory effect of JFK on H1975 cells was enhanced in a time-and dose-dependent manner,and the differences were statistically significant(all P<0.05);compared with the different concentrations of gefitinib alone for 48 h,the proliferation inhibition rate of H1975 cells after treatment with the corresponding concentration of gefitinib combined with JFK was more higher(all P<0.05).The differences in the glucose uptake of cells in each treatment group were not significantly different compared with the blank group(P>0.05),but the cellular lactate production in each treatment group was significantly lower than that in the blank group(P<0.05),and the combination group was significantly lower than the gefitinib group and JFK group(P<0.05);the activities of PFK and PK in the gefitinib group,those of HK and PFK in the JFK group,HK,PFK and PK in the combination group were significantly lower than those in the blank group(all P<0.05),and the activities of HK and PFK in the JFK group and the combination group were significantly l

关 键 词：H1975细胞 肺腺癌 金复康口服液 吉非替尼 有氧糖酵解 

分 类 号：R734.2[医药卫生—肿瘤]