Delineation of molecular mechanisms of sensitivity to lapatinib in breast cancer cell lines using global gene expression profiles
Delineation of molecular mechanisms of sensitivity to lapatinib in breast cancer cell lines using global gene expression profiles
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DOI：
10.1158/1535-7163.mct-05-0399
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发表时间：
2007-05-01
影响因子：
5.7
通讯作者：
Gilmer, Tona M.
中科院分区：
文献类型：
--
作者：
Hegde, Priti S.;Rusnak, David;Gilmer, Tona M.
拉帕替尼（GW 572016）是一种表皮生长因子受体（ErbB 1）和ErbB 2受体激酶活性的小分子双重抑制剂，目前正在进行III期临床试验。我们使用磷蛋白和微阵列分析进行靶向通路研究磷酸化和基因表达的变化，在人类乳腺癌细胞系中拉帕替尼的存在或不存在。在四种乳腺癌细胞系中进行了研究，其中两种对拉帕替尼有反应，两种对拉帕替尼无反应。反应性细胞系BT474和SKBr 3组成型过表达ErbB 2，拉帕替尼的IC 50分别为25或32 nmol/L。相比之下，无反应的MDA-MB-468和T47 D细胞表达低基础水平的ErbB 2，并显示IC 50值在微摩尔范围内。对拉帕替尼有反应的细胞对AKT途径中的多个基因表现出强烈的差异效应。暴露于1.0 μ mol/L拉帕替尼12 h后，在应答性BT474和SKBr 3细胞中，AKT 1、MAPK 9、HSPCA、IRAK 1和CCND 1转录物下调7- 25倍。相比之下，拉帕替尼在无应答乳腺癌细胞系中微弱下调AKT途径（途径中大多数基因下调< 5倍）。此外，由AKT负调控的促凋亡基因FOXO 3A在拉帕替尼应答的SKBr 3和BT474细胞中分别上调7倍和25倍。磷酸化Akt和Akt介导的FOXO 3A磷酸化也在暴露于拉帕替尼的响应性乳腺癌细胞系中降低。基因表达谱分析还显示拉帕替尼刺激雌激素和孕激素受体的表达，并调节参与细胞周期控制、糖酵解和脂肪酸代谢的基因的表达。在表达中等基础水平雌激素和孕激素受体的BT474和T47 D细胞中，1.0 μ mol/L拉帕替尼诱导表达7- 11倍。这些数据为拉帕替尼在乳腺癌细胞中的作用机制提供了见解。
Lapatinib (GW572016) is a small-molecule dual inhibitor of epidermal growth factor receptor (ErbB1) and ErbB2 receptor kinase activities currently in phase III clinical trials. We used phosphoprotein and microarray analyses to carry out targeted pathway studies of phosphorylation and gene expression changes in human breast cancer cell lines in the presence or absence of lapatinib. Studies were done in four breast cancer cell lines, two of which were responsive and two of which were nonresponsive to lapatinib. Responsive cell lines, BT474 and SKBr3, constitutively overexpress ErbB2 and show an IC50 of 25 or 32 nmol/L for lapatinib, respectively. In contrast, nonresponsive MDA-MB-468 and T47D cells expressed a low basal level of ErbB2 and showed IC50 values in the micromolar range. Cells responsive to lapatinib exhibited strong differential effects on multiple genes in the AKT pathway. After 12 h of exposure to 1.0 mu mol/L of lapatinib, AKT1, MAPK9, HSPCA, IRAK1, and CCND1 transcripts were down-regulated 7- to 25-fold in responsive BT474 and SKBr3 cells. In contrast, lapatinib weakly down-regulated the AKT pathway in nonresponsive breast cancer cell lines (< 5-fold down-regulation of most genes in the pathway). Furthermore, the proapoptotic gene FOXO3A, which is negatively regulated by AKT, was up-regulated 7- and 25-fold in lapatinib-responsive SKBr3 and BT474 cells, respectively. Phosphorylated Akt and Akt-mediated phosphorylation of FOXO3A also decreased in responsive breast cancer cell lines exposed to lapatinib. Gene expression profiling also revealed that lapatinib stimulated the expression of estrogen and progesterone receptors and modulated the expression of genes involved in cell cycle control, glycolysis, and fatty acid metabolism. In BT474 and T47D cells, which expressed moderate basal levels of the estrogen and progesterone receptors, 1.0 mu mol/L of lapatinib induced expression by 7- to 11- fold. These data provide insight into the mechanism of action of lapatinib in breast cancer cells.