人肺腺癌吉非替尼耐药细胞株中微小RNA的表达谱

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.

microRNA expression profiles in acquired gefitinib-resistant human lung adenocarcinoma cell line PC9/AB2 vs its parental cell line PC9

Objective: To identify the differential expression of microRNAs (miRNAs) in acquired gefitinib-resistant human lung adenocarcinoma cell line PC9/AB2 versus parental cell line PC9. Methods: The expression profiles of PC9/AB2 and PC9 cells were detected by using Agilent Human microRNA Microarray. The Agilent Feature Extraction software was used to analyze the differential expression of miRNA, and the Bioinformatics software was used to predict the potential target genes of differentially expressed miRNAs. The real-time fluorescenic quantitative-PCR (RFQ-PCR) was used to confirm the result of microRNA microarray. Results:There were thirty-six miRNAs up-regulated over 2-fold and twenty-four miRNAs down-regulated over 2-fold in PC9/AB2 cells as compared with those in PC9 cells. The result of microRNA microarray was validated by RFQ-PCR. The prediction result revealed that 16 miRNAs had their potential target genes, and 11 of which had the potential target genes over 100. Conclusion: The differential expression of miRNAs may be contributed to acquired gefitinib-resistance of human lung adenocarcinoma cells by regulating their target genes.