文章摘要
陈 君,黄佳丽,陈臻瑶,王朝霞,程志祥.非小细胞肺癌奥希替尼耐药细胞株的建立与鉴定及耐药后对化疗的敏感性[J].南京医科大学学报,2020,(8):1091~1097
非小细胞肺癌奥希替尼耐药细胞株的建立与鉴定及耐药后对化疗的敏感性
Establishment and identification of an osimertinib⁃resistant non⁃small cell lung cancer cell line and sensitivity to chemotherapy after drug resistance
投稿时间:2019-11-20  
DOI:10.7655/NYDXBNS20200803
中文关键词: 奥希替尼  非小细胞肺癌  耐药  表皮生长因子受体  化疗
英文关键词: osimertinib  non⁃small cell lung cancer  drug resistance  epidermal growth factor receptor  chemotherapy
基金项目:国家自然科学基金(81372395)
作者单位
陈 君 南京医科大学第二附属医院肿瘤医学中心江苏 南京 210011 
黄佳丽 南京医科大学第二附属医院肿瘤医学中心江苏 南京 210011 
陈臻瑶 南京医科大学第二附属医院肿瘤医学中心江苏 南京 210011 
王朝霞 南京医科大学第二附属医院肿瘤医学中心江苏 南京 210011 
程志祥 南京医科大学第二附属医院肿瘤医学中心江苏 南京 210011 
摘要点击次数: 217
全文下载次数: 167
中文摘要:
      目的:建立非小细胞肺癌奥希替尼耐药细胞株H1975AR,对亲本和耐药细胞株进行鉴定,探讨耐药机制。方法:在体外采用浓度递增的方法予奥希替尼处理诱导H1975细胞建立奥希替尼耐药细胞株H1975AR;光镜及Giemsa染色比较2种细胞形态学差异;第二代测序(next generation sequencing,NGS)检测表皮生长因子受体(epidermal growth factor receptor,EGFR)基因突变;磺酰罗丹明B(sulforthodamine B,SRB)法检测细胞增殖,评估细胞对EGFR?酪氨酸激酶抑制剂(tyrosine kinase inhibitor,TKI)和化疗药的敏感性;分别予0、10、100、1 000 nmol/L奥希替尼处理6 h,Western blot法检测EGFR及下游增殖信号的变化。结果:①SRB法测得H1975AR的耐药指数为3 634.75,显示H1975AR是奥希替尼耐药细胞株;②耐药细胞株形态发生改变且核浆比增大;③与H1975相比,给予奥希替尼处理对H1975AR的p?EGFR和p?ERK抑制作用不明显,且相同的给药浓度对H1975AR的p?AKT抑制作用较弱,同时下调c?Met的表达;④H1975和H1975AR对第一代EGFR?TKI耐药,对顺铂和紫杉醇敏感。结论:H1975AR细胞EGFR C797S顺式突变的产生是奥希替尼主要耐药机制,并对顺铂和紫杉醇敏感。
英文摘要:
      Objective:To establish an osimertinib?resistant non?small cell lung cancer cell line H1975AR,identify parental and drug?resistant cells,and explore the mechanism of drug resistance. Methods:Induction of H1975 by increasing the concentration of osimertinib in vitro was performed to establish osimertinib?resistant cell line H1975AR. The morphological differences between the two cells were compared by light microscopy and Giemsa staining. Next generation sequencing(NGS)was used to detect epidermal growth factor receptor(EGFR)mutations. Cell proliferation was determined by SRB assay to access sensitivity to EGFR?tyrosine kinase inhibitor(TKI) and chemotherapeutic drugs. Western blot detected the changes of EGFR and downstream proliferation signaling pathway after treating with 0、10、100、 1 000 nmol/L osimertinib for 6 h. Results:①The resistance index of H1975AR measured by SRB assay was 3 634.75,which indicated that H1975AR was an osimertinib?resistant cell line. ②The morphology of drug?resistant cell line changed and the nuclear?to?cytoplasmic ratio increased. ③Compared with H1975,the inhibitory effected on p?EGFR and p?ERK in H1975AR were not obvious,and the same dose concentration had a weak inhibitory effect on p?AKT in H1975AR,and the expression of c?Met was down?regulated. ④H1975 and H1975AR were resistant to the first?generation EGFR?TKI and sensitive to cisplatin and paclitaxel. Conclusion:The EGFR mutation of cis?C797S in H1975AR cells is the main mechanism of osimertinib resistance and H1975AR is sensitive to cisplatin and paclitaxel.
查看全文   查看/发表评论  下载PDF阅读器