Fernandez-Cobo Mariana, Holland James F, Pogo Beatriz G T
Department of Medicine, Mount Sinai School of Medicine, New York, New York, USA.
BMC Cancer. 2006 Apr 20;6:99. doi: 10.1186/1471-2407-6-99.
Searches for differentially expressed genes in tumours have made extensive use of array technology. Most samples have been obtained from tumour biopsies or from established tumour-derived cell lines. Here we compare cultures of non-immortalized breast cancer cells, normal non-immortalized breast cells and immortalized normal and breast cancer cells to identify which elements of a defined set of well-known cancer-related genes are differentially expressed.
Cultures of cells from pleural effusions or ascitic fluids from breast cancer patients (MSSMs) were used in addition to commercially-available normal breast epithelial cells (HMECs), established breast cancer cell lines (T-est) and established normal breast cells (N-est). The Atlas Human Cancer 1.2 cDNA expression array was employed. The data obtained were analysed using widely-available statistical and clustering software and further validated through real-time PCR.
According to Significance Analysis of Microarray (SAM) and AtlasImage software, 48 genes differed at least 2-fold in adjusted intensities between HMECs and MSSMs (p < 0.01). Some of these genes have already been directly linked with breast cancer, metastasis and malignant progression, whilst others encode receptors linked to signal transduction pathways or are otherwise related to cell proliferation. Fifty genes showed at least a 2.5-fold difference between MSSMs and T-est cells according to AtlasImage, 2-fold according to SAM. Most of these classified as genes related to metabolism and cell communication.
The expression profiles of 1176 genes were determined in finite life-span cultures of metastatic breast cancer cells and of normal breast cells. Significant differences were detected between the finite life-span breast cancer cell cultures and the established breast cancer cell lines. These data suggest caution in extrapolating information from established lines for application to clinical cancer research.
在肿瘤中寻找差异表达基因的研究广泛应用了阵列技术。大多数样本取自肿瘤活检组织或已建立的肿瘤衍生细胞系。在此,我们比较了非永生化乳腺癌细胞、正常非永生化乳腺细胞以及永生化正常和乳腺癌细胞的培养物,以确定一组已知的与癌症相关基因中哪些元件存在差异表达。
除了使用市售的正常乳腺上皮细胞(HMECs)、已建立的乳腺癌细胞系(T-est)和已建立的正常乳腺细胞(N-est)外,还使用了来自乳腺癌患者胸腔积液或腹水的细胞培养物(MSSMs)。采用了Atlas Human Cancer 1.2 cDNA表达阵列。使用广泛可用的统计和聚类软件对获得的数据进行分析,并通过实时PCR进一步验证。
根据微阵列显著性分析(SAM)和AtlasImage软件,HMECs和MSSMs之间有48个基因的校正强度差异至少为2倍(p < 0.01)。其中一些基因已直接与乳腺癌、转移和恶性进展相关,而其他基因编码与信号转导途径相关的受体或与细胞增殖有关。根据AtlasImage,MSSMs和T-est细胞之间有50个基因差异至少为2.5倍,根据SAM为2倍。这些基因大多数归类为与代谢和细胞通讯相关的基因。
在转移性乳腺癌细胞和正常乳腺细胞的有限寿命培养物中确定了1176个基因的表达谱。在有限寿命的乳腺癌细胞培养物和已建立的乳腺癌细胞系之间检测到显著差异。这些数据表明,在将来自已建立细胞系的信息外推应用于临床癌症研究时应谨慎。