Establishment of Afatinib-Resistant Non-Small Cell Lung Cancer Cells Derived from Tumor Xenograft Model [Afatinib treatment]. Homo sapiens

This experiment aimed to indentify signifcant expression genes in afatinib-resistant HCC827 cell lines under different treatment conditions. All the expression profiles of BR16-1 and BR16-2 cells were comapared to parental HCC827 cells. Overall design: When cell lines isolated from tumors grew at 70% confluence, these cells were treated with afatinib at 0, 10 ,100, and 1000 nM. After 48hr, total RNA was isolated by using Qiagen RNEasy mini kit. The RNA integrity and QC reports were carried based on manufacturer's manual. Labeled cRNA was hybridized to Illumina Human HT-12 Beadchips v4. Data were analysed by GeneSpring GX software (Agilent Techologies).

本实验旨在鉴定不同处理条件下耐阿法替尼（afatinib）的HCC827细胞系中的显著差异表达基因。将BR16-1与BR16-2细胞的全表达谱与亲本HCC827细胞进行比对。实验整体设计：当从肿瘤分离获得的细胞系汇合度达70%时，以浓度分别为0、10、100及1000 nM的阿法替尼处理各组细胞。培养48小时后，采用Qiagen RNEasy迷你试剂盒提取总RNA。依照试剂盒制造商的操作手册完成RNA完整性检测与质量控制（QC）分析。将标记后的互补RNA（cRNA）与Illumina Human HT-12 第4版微珠芯片进行杂交。数据采用GeneSpring GX软件（安捷伦科技，Agilent Technologies）进行分析。