Distinct effects of EGFR inhibitors on epithelial- and mesenchymal-like esophageal squamous cell carcinoma cells.
Distinct effects of EGFR inhibitors on epithelial- and mesenchymal-like esophageal squamous cell carcinoma cells.
复制标题
DOI：
10.1186/s13046-017-0572-7
复制
发表时间：
2017-08-01
期刊：
影响因子：
--
通讯作者：
Muto M
中科院分区：
文献类型：
--
作者：
Yoshioka M;Ohashi S;Ida T;Nakai Y;Kikuchi O;Amanuma Y;Matsubara J;Yamada A;Miyamoto S;Natsuizaka M;Nakagawa H;Chiba T;Seno H;Muto M
表皮生长因子受体（EGFR）在食管鳞状细胞癌（ESCC）的病理生理过程中起着关键作用。然而，EGFR抑制剂对ESCC的临床效果存在争议。本研究旨在确定决定EGFR抑制剂在ESCC细胞中治疗效果的因素。用EGFR抑制剂厄洛替尼或西妥昔单抗处理永生化人食管上皮细胞（EPC 2-hTERT）、转化人食管上皮细胞（T-Epi和T-Mes）和ESCC细胞（TE-1、TE-5、TE-8、TE-11、TE-11 R和HCE 4）。通过细胞计数或细胞周期分析评估对细胞生长的抑制作用。基因和蛋白质的表达水平，如外皮蛋白和细胞角蛋白13（鳞状细胞分化标志物），E-钙粘蛋白，波形蛋白进行了评估，通过实时聚合酶链反应或蛋白质印迹。为了检查间充质表型是否影响EGFR抑制剂的作用，我们用TGF-β1处理T-Epi细胞以建立间充质表型（间充质T-Epi细胞）。然后，我们比较了EGFR抑制剂对亲本T-Epi细胞和间充质T-Epi细胞的影响。用西妥昔单抗处理小鼠中TE-8（间充质样ESCC细胞）或TE-11 R（上皮样ESCC细胞）来源的异种移植肿瘤，并评价EGFR抑制剂的抗肿瘤作用。细胞分为上皮样或间充质样表型，确定的表达水平的E-钙粘蛋白和波形蛋白。厄洛替尼和西妥昔单抗均降低上皮样细胞而非间充质样细胞的细胞生长和细胞周期S期细胞的比例。此外，EGFR抑制剂诱导上皮样细胞而非间充质样细胞的鳞状细胞分化（定义为外皮蛋白和细胞角蛋白13的表达增加）。我们发现，EGFR抑制剂并不抑制间充质样细胞中EGFR的磷酸化，而在上皮样细胞中用EGFR抑制剂处理后观察到EGFR去磷酸化。此外，间充质T-Epi细胞通过规避EGFR信号的去磷酸化而显示出对EGFR抑制剂的抗性。西妥昔单抗在TE-11 R（上皮样）来源的异种移植肿瘤中持续显示抗肿瘤作用，并增加了外皮蛋白表达，但在TE-8（间充质样）来源的异种移植肿瘤中没有。决定EGFR抑制剂在ESCC细胞中的治疗效果的因素是代表上皮样或间充质样细胞的表型。间质样ESCC细胞对EGFR抑制剂具有抗性，因为EGFR信号传导未被阻断。EGFR抑制剂对上皮样ESCC细胞显示出抗肿瘤作用，并伴有促进鳞状细胞分化。本文的在线版本（doi：10.1186/s13046-017-0572-7）包含补充材料，可供授权用户使用。
Epidermal growth factor receptor (EGFR) plays a pivotal role in the pathophysiology of esophageal squamous cell carcinoma (ESCC). However, the clinical effects of EGFR inhibitors on ESCC are controversial. This study sought to identify the factors determining the therapeutic efficacy of EGFR inhibitors in ESCC cells. Immortalized-human esophageal epithelial cells (EPC2-hTERT), transformed-human esophageal epithelial cells (T-Epi and T-Mes), and ESCC cells (TE-1, TE-5, TE-8, TE-11, TE-11R, and HCE4) were treated with the EGFR inhibitors erlotinib or cetuximab. Inhibitory effects on cell growth were assessed by cell counting or cell-cycle analysis. The expression levels of genes and proteins such as involucrin and cytokeratin13 (a squamous differentiation marker), E-cadherin, and vimentin were evaluated by real-time polymerase chain reaction or western blotting. To examine whether mesenchymal phenotype influenced the effects of EGFR inhibitors, we treated T-Epi cells with TGF-β1 to establish a mesenchymal phenotype (mesenchymal T-Epi cells). We then compared the effects of EGFR inhibitors on parental T-Epi cells and mesenchymal T-Epi cells. TE-8 (mesenchymal-like ESCC cells)- or TE-11R (epithelial-like ESCC cells)-derived xenograft tumors in mice were treated with cetuximab, and the antitumor effects of EGFR inhibitors were evaluated. Cells were classified as epithelial-like or mesenchymal-like phenotypes, determined by the expression levels of E-cadherin and vimentin. Both erlotinib and cetuximab reduced cell growth and the ratio of cells in cell-cycle S phase in epithelial-like but not mesenchymal-like cells. Additionally, EGFR inhibitors induced squamous cell differentiation (defined as increased expression of involucrin and cytokeratin13) in epithelial-like but not mesenchymal-like cells. We found that EGFR inhibitors did not suppress the phosphorylation of EGFR in mesenchymal-like cells, while EGFR dephosphorylation was observed after treatment with EGFR inhibitors in epithelial-like cells. Furthermore, mesenchymal T-Epi cells showed resistance to EGFR inhibitors by circumventing the dephosphorylation of EGFR signaling. Cetuximab consistently showed antitumor effects, and increased involucrin expression in TE-11R (epithelial-like)-derived xenograft tumors but not TE-8 (mesenchymal-like)-derived xenograft tumors. The factor determining the therapeutic effects of EGFR inhibitors in ESCC cells is the phenotype representing the epithelial-like or mesenchymal-like cells. Mesenchymal-like ESCC cells are resistant to EGFR inhibitors because EGFR signaling is not blocked. EGFR inhibitors show antitumor effects on epithelial-like ESCC cells accompanied by promotion of squamous cell differentiation. The online version of this article (doi:10.1186/s13046-017-0572-7) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
21.3
作者：
Kolev, Vihren;Mandinova, Anna;Guinea-Viniegra, Juan;Hu, Bing;Lefort, Karine;Lambertini, Chiara;Neel, Victor;Dummer, Reinhard;Wagner, Erwin F.;Dotto, G. Paolo
通讯作者：
Dotto, G. Paolo
影响因子：
29.4
作者：
Ohashi S;Natsuizaka M;Yashiro-Ohtani Y;Kalman RA;Nakagawa M;Wu L;Klein-Szanto AJ;Herlyn M;Diehl JA;Katz JP;Pear WS;Seykora JT;Nakagawa H
通讯作者：
Nakagawa H
影响因子：
11.2
作者：
Ohashi S;Natsuizaka M;Wong GS;Michaylira CZ;Grugan KD;Stairs DB;Kalabis J;Vega ME;Kalman RA;Nakagawa M;Klein-Szanto AJ;Herlyn M;Diehl JA;Rustgi AK;Nakagawa H
通讯作者：
Nakagawa H
影响因子：
4.6
作者：
Kikuchi O;Ohashi S;Horibe T;Kohno M;Nakai Y;Miyamoto S;Chiba T;Muto M;Kawakami K
通讯作者：
Kawakami K
影响因子：
3.7
作者：
Ohashi S;Kikuchi O;Tsurumaki M;Nakai Y;Kasai H;Horimatsu T;Miyamoto S;Shimizu A;Chiba T;Muto M
通讯作者：
Muto M