A stable explant culture of HER2/neu invasive carcinoma supported by alpha-Smooth Muscle Actin expressing stromal cells to evaluate therapeutic agents.
A stable explant culture of HER2/neu invasive carcinoma supported by alpha-Smooth Muscle Actin expressing stromal cells to evaluate therapeutic agents.
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DOI：
10.1186/1471-2407-8-119
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发表时间：
2008-04-24
期刊：
影响因子：
3.8
通讯作者：
Piechocki MP
中科院分区：
文献类型：
--
作者：
Piechocki MP
为了更好地了解治疗药物对浸润性癌症中肿瘤微环境的影响，我们从浸润性小叶癌中开发了一种共培养模型。表达HER 2/neu的肿瘤细胞在表达α-平滑肌肌动蛋白（α-SMA）的肿瘤间质包围的巢中组织，类似于侵袭性肿瘤的形态。该共培养物，即乳腺腺癌模型（MAM-1），保持HER 2/neu阳性肿瘤细胞与α-SMA反应性基质细胞的比例为1：1，并在体外更新该配置超过20代。我们通过微阵列分析和免疫细胞化学表征了MAM-1模型的细胞成分。我们开发了流式细胞术检测来评估肿瘤和间质对酪氨酸激酶抑制剂易瑞沙的相对反应。MAM-1基因表达谱包含代表ErbB-2乳腺癌特征的簇和与浸润性乳腺癌相关的基质特异性簇。该模型的稳定性和抗原标记肿瘤和间质组分的能力使我们能够确定易瑞沙（一种受体酪氨酸激酶抑制剂）靶向肿瘤细胞群的特异性。处理导致肿瘤细胞中磷酸化pMEK 1/2和pp 44/42 MAPK的选择性剂量依赖性降低。在24小时内，肿瘤细胞分数减少1.9倍，而基质细胞分数增加>3倍，这与肿瘤细胞中磷酸-pp 44/42 MAPK、MEK 1/2和PCNA的特异性减少以及基质细胞中的相互增加一致。肿瘤细胞巢的侵蚀和基质细胞的生长增强类似于纤维化反应。该模型证明了易瑞沙对表达HER 2/neu的肿瘤细胞相对于肿瘤相关肌成纤维细胞的特异性，并且适用于描述治疗对乳腺肿瘤微环境中信号转导的影响，并改善可以双重或差异靶向微环境中肿瘤和基质成分的策略。
To gain a better understanding of the effects of therapeutic agents on the tumor microenvironment in invasive cancers, we developed a co-culture model from an invasive lobular carcinoma. Tumor cells expressing HER2/neu organize in nests surrounded by alpha-Smooth Muscle Actin (α-SMA) expressing tumor stroma to resemble the morphology of an invading tumor. This co-culture, Mammary Adenocarcinoma Model (MAM-1) maintains a 1:1 ratio of HER2/neu positive tumor cells to α-SMA-reactive stromal cells and renews this configuration for over 20 passages in vitro. We characterized the cellular elements of the MAM-1 model by microarray analysis, and immunocytochemistry. We developed flow cytometric assays to evaluate the relative responses of the tumor and stroma to the tyrosine kinase inhibitor, Iressa. The MAM-1 gene expression profile contains clusters that represent the ErbB-2 breast cancer signature and stroma-specific clusters associated with invasive breast cancers. The stability of this model and the ability to antigenically label the tumor and stromal fractions allowed us to determine the specificity of Iressa, a receptor tyrosine kinase inhibitor, for targeting the tumor cell population. Treatment resulted in a selective dose-dependent reduction in phospho-pMEK1/2 and pp44/42MAPK in tumor cells. Within 24 h the tumor cell fraction was reduced 1.9-fold while the stromal cell fraction increased >3-fold, consistent with specific reductions in phospho-pp44/42 MAPK, MEK1/2 and PCNA in tumor cells and reciprocal increases in the stromal cells. Erosion of the tumor cell nests and augmented growth of the stromal cells resembled a fibrotic response. This model demonstrates the specificity of Iressa for HER2/neu expressing tumor cells versus the tumor associated myofibroblasts and is appropriate for delineating effects of therapy on signal transduction in the breast tumor microenvironment and improving strategies that can dually or differentially target the tumor and stromal elements in the microenvironment.
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影响因子：
24.3
作者：
Inoue, T;Fujishima, S;Yamaguchi, K
通讯作者：
Yamaguchi, K
影响因子：
6.4
作者：
Piechocki, Marie P.;Dibbley, Susan K.;Yoo, George H.
通讯作者：
Yoo, George H.
影响因子：
8
作者：
Landis, MD;Seachrist, DD;Keri, RA
通讯作者：
Keri, RA
影响因子：
11.2
作者：
Piechocki, Marie P.;Yoo, George H.;Lonardo, Fulvio
通讯作者：
Lonardo, Fulvio
DOI：
10.1073/pnas.0511200103
发表时间：
2006-09-19
影响因子：
11.1
作者：
Sakai, Norihiko;Wada, Takashi;Kaneko, Shuichi
通讯作者：
Kaneko, Shuichi