糖酵解在非小细胞肺癌吉非替尼耐药中的作用研究  

Effects of glycolysis in Gefitinib resistance in non⁃small cell lung cancer

作　　者：李楠[1] 路晨阳 祝阿妮 杨栋才 马倩[1] 严奉奇 贾卫红 LI Nan;LU Chenyang;ZHU Ani;YANG Dongcai;MA Qian;YAN Fengqi;JIA Weihong(Department of Respiratory and Critical Care Medicine,Xi′an NO.3 Hospital,the Affiliated Hospital of Northwest University,Xi′an 710083,China;Department of Urology,Tang Du Hospital,the Air Force Military Medical University,Xi′an 710038,China)

机构地区：[1]西北大学附属医院西安市第三医院呼吸与危重症医学科,西安710083 [2]空军军医大学唐都医院泌尿外科,西安710038

出　　处：《中国癌症防治杂志》2024年第1期50-55,共6页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT

基　　金：西安市卫生健康委员会科研一般项目(2021lyb41)。

摘　　要：目的探讨糖酵解在非小细胞肺癌(non-small cell lung cancer,NSCLC)吉非替尼(Gefitinib)耐药中的变化及作用。方法体外培养NSCLC亲本细胞PC-9和A549,并采用浓度递增法构建NSCLC吉非替尼耐药细胞PC-9-GR和A549-GR,通过CCK-8和平板克隆形成实验鉴定细胞耐药性,葡萄糖摄取检测试剂盒和乳酸测试盒分别检测葡萄糖代谢和乳酸产出水平,qRT-PCR和Western blot实验检测细胞糖酵解关键酶的mRNA和蛋白表达水平。采用糖酵解抑制剂2-脱氧-D-葡萄糖(2-deoxy-D-glucose,2-DG)抑制细胞糖酵解后,观察细胞活力及吉非替尼耐药性的变化情况。结果相较于各自亲本细胞,耐药细胞PC-9-GR和A549-GR对吉非替尼抵抗性增强(均P<0.01),葡萄糖代谢速率和乳酸产出水平提高(均P<0.01);糖酵解关键酶HK2、LDHA和PFK的mRNA表达水平升高(均P<0.001),HK2、LDHA、PFKP和PKM2的蛋白表达水平增加(均P<0.001)。2-DG对耐药细胞活力的抑制作用较各自亲本细胞更明显(均P<0.05),且在一定程度上可增强吉非替尼对耐药细胞的杀伤效应。结论NSCLC吉非替尼耐药伴随着糖酵解的激活,该过程可能与糖酵解关键酶表达升高有关,抑制其水平或活性可能是逆转吉非替尼耐药的有效措施。Objective To investigate the changes and effects of glycolysis in Gefitinib resistance in non‑small cell lung cancer(NSCLC).Methods NSCLC parental cells PC‑9 and A549 were cultured in vitro,and NSCLC Gefitinib‑resistant cells PC‑9‑GR and A549‑GR were constructed by the increasing concentration method.Cell resistance was identified by CCK‑8 and colony formation assay.The glucose metabolism and lactate production levels were detected using glucose and lactate assay kits,respectively.qRT‑PCR and Western blot were used to detect the mRNA and protein levels of the key enzymes in glycolysis.After inhibition of glycolysis by 2‑deoxy‑D‑glucose,the viability of cells and the changes of drug resistance to Gefitinib were observed.Results Compared with parental cells,the resistance of PC‑9‑GR and A549‑GR cells to Gefitinib was enhanced(all P<0.01),and glucose metabolism rate and lactate production level were increased(all P<0.01).The mRNA expression levels of HK2,LDHA and PFK and the protein levels of HK2,LDHA,PFKP and PKM2 were increased(all P<0.001).The inhibitory effect of 2‑DG on the activities of drug‑resistant cells was more significant than on those of their respective parental cells(all P<0.05),and to some extent,it could enhance the killing effect of Gefitinib on drug‑resistant cells.Conclusions Gefitinib resistance in NSCLC is accompanied by the activation of glycolysis,and is associated with increased expression of key enzymes in glycolysis.Inhibiting the levels or activities of glycolysis may be an effective measure to reverse the drug resistance of Gefitinib.

关 键 词：非小细胞肺癌 吉非替尼 耐药 糖酵解 代谢重编程 

分 类 号：R734.2[医药卫生—肿瘤]