E-Cadherin and EpCAM expression by NSCLC tumour cells associate with normal fibroblast activation through a pathway initiated by integrin αvβ6 and maintained through TGFβ signalling
E-Cadherin and EpCAM expression by NSCLC tumour cells associate with normal fibroblast activation through a pathway initiated by integrin αvβ6 and maintained through TGFβ signalling
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DOI：
10.1038/onc.2013.600
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发表时间：
2015-02-05
期刊：
影响因子：
8
通讯作者：
Barry, S. T.
中科院分区：
文献类型：
--
作者：
Eberlein, C.;Rooney, C.;Barry, S. T.
肿瘤基质中的成纤维细胞（癌症相关成纤维细胞）影响肿瘤进展和对治疗的反应;关于肿瘤细胞吸收正常成纤维细胞的机制知之甚少。为了研究肿瘤细胞对成纤维细胞的活化，将一组非小细胞肺癌（NSCLC）细胞系和正常人皮肤成纤维细胞共培养。NSCLC细胞的一个亚组诱导活化的癌症相关成纤维细胞样成纤维细胞表型，其通过诱导成纤维细胞α-平滑肌肌动蛋白表达来定义。激活成纤维细胞的肿瘤细胞与E-钙粘蛋白和EpCAM表达以及整合素α v β 6的表达相关。活化肿瘤细胞与成纤维细胞的共培养导致诱导与肿瘤细胞侵袭和生长相关的转录物TGF β 1和TGFBR 1、SERPINE-1、BMP 6、SPHK 1和MMP 9。成纤维细胞活化被α v β 6/8整联蛋白阻断抗体（264 RAD）和TGF-β I型受体活化素样激酶（ALK 5）的小分子抑制剂（SB 431542）抑制，证明TGF β途径的反式激活启动成纤维细胞活化。整联蛋白和ALK 5拮抗剂均抑制起始。当在共培养3天后添加时，仅ALK 5有效。这表明，尽管活化是α v β 6依赖性的，但一旦成纤维细胞被活化，替代性TGF β途径调节剂维持活化环。与单一培养相比，在共培养中，活化细胞对司美替尼、AZD 8931和阿法替尼的敏感性降低。相比之下，非活化细胞在单培养和共培养中对司美替尼和AZD 8931均不敏感。总之，NSCLC细胞系，E-钙粘蛋白，EpCAM和α v β 6表达阳性，通过α v β 6/TGF β信号在体外激活正常成纤维细胞，并影响基因表达和对治疗剂的反应。
Fibroblasts in the tumour stroma (cancer-associated fibroblasts) influence tumour progression and response to therapeutics; little is known about the mechanisms through which the tumour cell co-opts a normal fibroblast. To study the activation of fibroblasts by tumour cells, a panel of non-small cell lung cancer (NSCLC) cell lines and normal human dermal fibroblasts were co-cultured. A subset of the NSCLC cells induced an activated cancer-associated fibroblast-like fibroblast phenotype defined by induction of fibroblast alpha-smooth muscle actin expression. Tumour cells that activated fibroblasts were associated with E-Cadherin and EpCAM expression and expression of integrin alpha v beta 6. Co-culture of activating tumour cells with fibroblasts resulted in induction of transcripts associated with tumour cell invasion and growth, TGF beta 1 and TGFBR1, SERPINE-1, BMP6, SPHK1 and MMP9. Fibroblast activation was inhibited by an alpha v beta 6/8 integrin blocking antibody (264RAD) and a small molecule inhibitor of the TGF-beta type I receptor activin-like kinase (ALK5) (SB431542), demonstrating that transactivation of the TGF beta pathway initiates fibroblast activation. Both integrin and ALK5 antagonists inhibited initiation. Only ALK5 was effective when added after 3 days of co-culture. This suggests that although activation is alpha v beta 6-dependent, once fibroblasts are activated alternative TGF beta pathway regulators maintain an activation loop. In co-culture activating cells had reduced sensitivity to selumetinib, AZD8931 and afatinib compared with mono-culture. In contrast, non-activating cells were insensitive to selumetinib and AZD8931 in both mono-culture and co-culture. In conclusion NSCLC cell lines, positive for E-Cadherin, EpCAM and alpha v beta 6 expression, activate normal fibroblasts through alpha v beta 6/TGF beta signalling in vitro, and influence both gene expression and response to therapeutic agents.