基因芯片筛选CD133^+/CD133^-肺腺癌细胞中新的耐药基因

Screening and Identification of Novel Drug-resistant Genes in CD133^+ and CD133^- Lung Adenosarcoma Cells Using cDNA Microarray

背景与目的肿瘤干细胞可能是肿瘤多药耐药的主要原因,CD133是目前较为公认的肿瘤干细胞标记物。本研究旨在应用功能分类基因芯片筛选CD133+和CD133-肺腺癌细胞中差异表达的肿瘤耐药基因,寻求新的肺癌耐药相关基因。方法免疫磁珠分选法分选A549细胞,采用功能分类基因芯片筛选CD133+和CD133-肺腺癌细胞中差异表达的肿瘤耐药基因,并使用RT-qPCR验证。顺铂半数有效抑制浓度(half inhibiting concentration,IC50)、阿霉素IC50作用A549细胞48 h后,RT-qPCR检测肿瘤耐药基因CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα和PPARβ/δ的表达变化。结果共筛查出31个差异表达的肿瘤耐药基因,与CD133-细胞相比,CD133+细胞有30个基因表达上调,1个基因表达下调。RT-qPCR结果与芯片一致。A549细胞经1.97μg/mL顺铂或0.61μg/mL阿霉素作用48 h后,CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα和PPARβ/δ等肿瘤耐药基因表达上调。结论利用功能分类基因芯片筛选出31个可能与CD133+肺腺癌细胞耐药相关的基因,其中CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα和PPARβ/δ为新发现的肺癌耐药相关基因。

Background and objective Cancer stem cells （CSCs） are responsible for multi-drug resistance in tu-mors. CD133 is a known biomarker of CSCs. hTe aim of this study is to screen for drug-resistant differentially expressed genes in CD133＋and CD133-lung cancer cells and to identify novel lung tumor drug-resistant genes. Methods Magnetic activated cell sorting was used to isolate CD133＋and CD133-cells from human lung cancer cell line A549, and drug-resistant microarray was used to detect drug-resistant genes in the these cells. RT-qPCR was used to examine the expression of six lung tumor drug-resistant genes in pre-and post-chemotherapeutic A549 cells. Results A total of 31 differentially expressed genes were screened by microar-ray analysis. Of these genes, 30 were upregulated and one was downregulated in CD133＋cells compared with CD133-cells. Re-sults were veriifed by RT-qPCR. CYP2C19, CYP2D6, CYP2E1, GSK3α, PPARα, and PPARβ/δwere signiifcantly upregulated atfer the A549 cells were treated with 1.97μg/mL DDP or 0.61μg/mL doxorubicin for 48 h. Conclusion hTe drug resistance of lung adenosarcoma may be correlated with 31 differentially expressed genes screened by drug-resistant microarray. CYP2C19, CYP2D6, CYP2E1, GSK3α, PPARα, and PPARβ/δmight be novel lung adenosarcoma drug-resistant genes.