吉非替尼耐药肺癌细胞株建立及其EGFR信号通路改变  

Establishment of gefitinib resistant lung cancercell line and changes of EGFR signaling pathway

作　　者：丁萌 廖海秀 周楠楠 程娟[1] 王琴[1] 管世鹤[1] 周强[1] 胡伟[1] 陈礼文[1] DING Meng;LIAO Hai-xiu;ZHOU Nan-nan;CHENG Juan;WANG Qin;GUAN Shi-he;ZHOU Qiang;HU Wei;CHEN Li-wen(Dept of Laboratory, the Second Hospital of Anhui Medical University, Hefei 230601, China)

机构地区：[1]安徽医科大学第二附属医院检验科,安徽合肥230601

出　　处：《中国药理学通报》2020年第5期634-639,共6页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金资助项目(No 180805MH229)。

摘　　要：目的建立吉非替尼(gefitinib)耐药的表皮生长因子受体(epidermal growth factor receptor,EGFR)突变肺腺癌H3255和HCC827细胞株并探究其EGFR下游信号通路改变。方法采用Gefitinib浓度梯度递增法诱导建立H3255和HCC827耐药株;CCK8法检测比较Gefitinib耐药细胞株H3255/GR和HCC827/GR的增殖;Western blot检测比较H3255、HCC827、H3255/GR、HCC827/GR细胞的EGFR下游信号的改变。结果 H3255/GR和HCC827/GR细胞的增殖显著低于非耐药株;H3255/GR和HCC827/GR耐药细胞株磷酸化信号分子p-AKT,p-ERK1/2和p-Stat3与其未耐药株相比均未观察到明显改变。Gefitinib处理的H3255/GR细胞p-Stat3和p-ERK1/2表达水平亦无显著改变,但p-AKT表达显著下调,而吉非替尼仅轻微抑制未耐药株p-ERK1/2表达;在HCC827/GR细胞,p-AKT和p-STAT3不被吉非替尼抑制,p-ERK1/2则被中度抑制。结论H3255/GR和HCC827/GR细胞株Gefitinib耐药的信号机制存在明显差异。Aim To establish gefitinib resistant epidermal growth factor receptor(EGFR)mutant lung cancer cell lines and to explore the changes of downstream signaling pathway of EGFR.Methods Gefitinib concentration gradient method was used to induce the establishment of H3255 and HCC827 resistant cell lines,and CCK8 assay was used to detect the proliferation of gefitinib resistant cell lines H3255/GR and HCC827/GR.Western blot was used to detect the changes of EGFR downstream signals in H3255,HCC827,H3255/GR and HCC827/GR cells.Results The proliferation of H3255/GR and HCC827/GR cells was significantly lower than that of non-drug resistant cells.The phosphorylation signal molecules p-AKT,p-ERK1/2 and p-STAT3 of H3255/GR and HCC827/GR drug-resistant cell lines were no significantly changed compared with their non-drug-resistant cell lines.There was no significant change in the expression of p-STAT3 and p-ERK1/2 in H3255/GR cells treated with gefitinib,but the expression of p-AKT was significantly down-regulated,while gefitinib only slightly inhibited the expression of p-ERK1/2 in drug-resistant H3255/GR cells.In HCC827/GR cells,p-AKT and p-STAT3 were not inhibited by gefitinib,while p-ERK1/2 was moderately inhibited.Conclusion There are significant differences in the signal mechanism of drug resistance between H3255/GR and HCC827/GR cell lines.

关 键 词：表皮生长因子受体 肺癌 吉非替尼 耐药 ERK/p-ERK AKT/p-AKT STAT3/p-STAT3 

分 类 号：R392.24[医药卫生—免疫学] R392.11[医药卫生—基础医学]