Shen Dejun, Chang Helena R, Chen Zugen, He Jianbo, Lonsberry Victor, Elshimali Yahya, Chia David, Seligson David, Goodglick Lee, Nelson Stanley F, Gornbein Jeffrey A
Gonda/UCLA Breast Cancer Research Laboratory, Department of Surgery, Revlon/UCLA Breast Center, University of California at Los Angeles, David Geffen School of Medicine, Los Angeles, CA 90095, USA.
Biochem Biophys Res Commun. 2005 Jan 7;326(1):218-27. doi: 10.1016/j.bbrc.2004.10.214.
To test the efficacy of combined high-throughput analyses (HTA) in target gene identification, screening criteria were set using >fivefold difference by microarray and statistically significant changes (p<0.01) in SAGE and EST. Microarray analysis of two normal and seven breast cancer samples found 129 genes with >fivefold changes. Further SAGE and EST analyses of these genes identified four qualified genes, ERBB2, GATA3, AGR2, and ANXA1. Their expression pattern was validated by RT-PCR in both breast cell lines and tissue samples. Loss of ANXA1 in breast cancer was further confirmed at mRNA level by Human Breast Cancer Tissue Profiling Array and at protein level by immunohistochemical staining. This study demonstrated that combined HTA effectively narrowed the number of genes for further study, while retaining the sensitivity in identifying biologically important genes such as ERBB2 and ANXA1. A distinctive loss of ANXA1 in breast cancer suggests its involvement in maintaining normal breast biology.
为了测试联合高通量分析（HTA）在靶基因鉴定中的功效，使用微阵列分析中大于五倍的差异以及SAGE和EST中具有统计学意义的变化（p<0.01）来设定筛选标准。对两个正常样本和七个乳腺癌样本进行微阵列分析，发现了129个变化大于五倍的基因。对这些基因进一步进行SAGE和EST分析，鉴定出四个合格基因，即ERBB2、GATA3、AGR2和ANXA1。它们的表达模式通过RT-PCR在乳腺细胞系和组织样本中得到验证。通过人乳腺癌组织芯片在mRNA水平以及通过免疫组化染色在蛋白质水平进一步证实了乳腺癌中ANXA1的缺失。本研究表明，联合HTA有效地减少了进一步研究的基因数量，同时保留了识别诸如ERBB2和ANXA1等生物学重要基因的敏感性。乳腺癌中ANXA1的明显缺失表明其参与维持正常乳腺生物学功能。
