应用基因芯片技术筛选乳腺癌阿霉素耐药相关基因表达谱

Screening the differential expression of adriamycin-resistance related genes of breast cancer by cDNA microarray

目的应用基因芯片技术研究人乳腺癌阿霉素耐药细胞株MCF-7／ADR与其亲本细胞株MCF-7基因表达谱的差异，筛选阿霉素耐药相关基因。方法选择人乳腺癌阿霉素耐药细胞株MCF-7／ADR与其亲本细胞株MCF-7为研究对象，应用四甲基偶氮唑盐（MTT）快速比色法检测、比较阿霉素对MCF-7／ADR细胞与MCF-7细胞的体外抑制率，然后应用含14755个基因的人cDNA基因芯片检测MCF-7／ADR细胞与MCF-7细胞基因表达谱的差异，筛选阿霉素耐药相关基因。结果阿霉素对MCF-7／ADR细胞的体外抑制率明显低于MCF-7细胞（P〈0．05），MCF-7／ADR细胞对阿霉素耐药性明显强于MCF-7细胞。MCF-7／ADR细胞与MCF-7细胞中表达差异的基因2374个，MCF-7／ADR细胞较MCF-7细胞表达上调的基因1099个，表达下调的基因1275个；10倍以上上调表达的基因99个，10倍以上下调表达的基因71个。表达显著上调的基因为Bcl-2、GSTP1、c—myc、MMP-1、NNMT，表达显著下调的基因为p53、p21、p27、CYPIA1。结论乳腺癌阿霉素耐药是一个多基因、多环节、多途径参与的过程，涉及多种基因表达的变化。运用基因芯片技术检测乳腺癌阿霉素耐药基因，有望为指导临床选择最佳个体化化疗方案开辟新途径。

Objective To study the differential gene expression of adriamycin-resistance of breast cancer cell line MCF-7/ADR as compared with its parent cell MCF-7 and screen the related genes. Methods The in vitro inhibitory rate of MCF-7/ ADR and MCF-7 cell lines to ADM （adriamycin） was examined by MTF assay. Then a cDNA microamay representing 14 755 genes was used to analyze the expression profiles of these two breast cancer cell lines. Results The inhibitory rate of MCF-7/ADR to ADM was lower than that of MCF-7 （ P 〈 0. 05 ） and the drug resistance to ADR of MCF-7/ADR was stronger than that of MCF-7. There were 2374 differential expression genes in MCF-7/adr and MCF-7 cell lines, of which 1099 genes upregulated with 99 genes 10-fold upregulated and 1275 genes downregulated with 71 genes 10-fold downregulated. The important upregulated genes were Bcl-2, GSTP1, c-myc, MMP-1 and NNMT while those downregulated ones p53, p21, p27 and CYPIA1. Conclusion Drug resistance of breast cancer is a complicated process involving the expresison of many upregulated or downregulated genes. The application of gene chip in screening drug-resistant genes may be a new approach for selecting the optimal chemotherapeutic regimens.