中国肺癌杂志
中國肺癌雜誌
중국폐암잡지
CHINESE JOURNAL OF LUNG CANCER
2014年
6期
437-443
,共7页
王红艳%郑少秋%涂永生%张雅洁
王紅豔%鄭少鞦%塗永生%張雅潔
왕홍염%정소추%도영생%장아길
CD133%肺肿瘤%耐药%肿瘤耐药基因芯片
CD133%肺腫瘤%耐藥%腫瘤耐藥基因芯片
CD133%폐종류%내약%종류내약기인심편
CD133%Lung neoplasms%Multi-drug resistant%Drug-resistant 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。
揹景與目的腫瘤榦細胞可能是腫瘤多藥耐藥的主要原因,CD133是目前較為公認的腫瘤榦細胞標記物。本研究旨在應用功能分類基因芯片篩選CD133+和CD133-肺腺癌細胞中差異錶達的腫瘤耐藥基因,尋求新的肺癌耐藥相關基因。方法免疫磁珠分選法分選A549細胞,採用功能分類基因芯片篩選CD133+和CD133-肺腺癌細胞中差異錶達的腫瘤耐藥基因,併使用RT-qPCR驗證。順鉑半數有效抑製濃度(half inhibiting concentration, IC50)、阿黴素IC50作用A549細胞48 h後,RT-qPCR檢測腫瘤耐藥基因CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα和PPARβ/δ的錶達變化。結果共篩查齣31箇差異錶達的腫瘤耐藥基因,與CD133-細胞相比,CD133+細胞有30箇基因錶達上調,1箇基因錶達下調。RT-qPCR結果與芯片一緻。A549細胞經1.97μg/mL順鉑或0.61μg/mL阿黴素作用48 h後,CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα和PPARβ/δ等腫瘤耐藥基因錶達上調。結論利用功能分類基因芯片篩選齣31箇可能與CD133+肺腺癌細胞耐藥相關的基因,其中CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα和PPARβ/δ為新髮現的肺癌耐藥相關基因。
배경여목적종류간세포가능시종류다약내약적주요원인,CD133시목전교위공인적종류간세포표기물。본연구지재응용공능분류기인심편사선CD133+화CD133-폐선암세포중차이표체적종류내약기인,심구신적폐암내약상관기인。방법면역자주분선법분선A549세포,채용공능분류기인심편사선CD133+화CD133-폐선암세포중차이표체적종류내약기인,병사용RT-qPCR험증。순박반수유효억제농도(half inhibiting concentration, IC50)、아매소IC50작용A549세포48 h후,RT-qPCR검측종류내약기인CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα화PPARβ/δ적표체변화。결과공사사출31개차이표체적종류내약기인,여CD133-세포상비,CD133+세포유30개기인표체상조,1개기인표체하조。RT-qPCR결과여심편일치。A549세포경1.97μg/mL순박혹0.61μg/mL아매소작용48 h후,CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα화PPARβ/δ등종류내약기인표체상조。결론이용공능분류기인심편사선출31개가능여CD133+폐선암세포내약상관적기인,기중CYP2C19、CYP2D6、CYP2E1、GSK3α、PPARα화PPARβ/δ위신발현적폐암내약상관기인。
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.