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目的:分析人肺腺癌吉非替尼耐药细胞株PC9/AB2与亲本细胞株PC9中微小RNA(microRNA,miRNA)表达谱的差异。方法:应用Agilent human miRNA芯片分别检测PC9/AB2细胞与PC9细胞miRNA的表达谱,随后采用Agilent Feature Extraction软件分析并筛选差异表达的miRNA,应用生物信息软件预测筛选出的差异表达miRNA的潜在靶基因。应用实时荧光定量-PCR(real-time fluorogenic quantitative-PCR,RFQ-PCR)验证芯片检测结果。结果:与PC9细胞比较,PC9/AB2细胞中有36个miRNA表达上调>2倍,有24个miRNA表达下调>2倍。RFQ-PCR验证了芯片的结果。软件预测发现,16个miRNA有潜在靶基因,其中11个miRNA的靶基因>100个。结论:筛选获得的差异表达miRNA可能通过调控其靶基因而参与人肺腺癌细胞对吉非替尼的耐药。
OBJECTIVE: To analyze the differential expression profiles of microRNA (miRNA) in the human lung adenocarcinoma cell line gefitinib, PC9 / AB2 and the parent cell line PC9. Methods: The miRNA expression profiles of PC9 / AB2 cells and PC9 cells were detected by Agilent human miRNA microarray. The differentially expressed miRNAs were analyzed and screened by Agilent Feature Extraction software. The bioinformatics software was used to predict the potential target genes of differentially expressed miRNAs . Real-time fluorogenic quantitative-PCR (RFQ-PCR) was used to validate the detection results. Results: Compared with PC9 cells, 36 miRNAs were upregulated> 2 fold in PC9 / AB2 cells and 24 miRNAs were downregulated> 2 fold. RFQ-PCR verified the results of the chip. Software predicts that 16 miRNAs have potential target genes, of which 11 target miRNAs have> 100 target genes. Conclusion: The differentially expressed miRNAs obtained by screening may participate in the resistance of gefitinib to human lung adenocarcinoma cells by regulating their target genes.