EGFR inhibitor AG1478 blocks the formation of 3D structures mainly through ERK signaling pathway in Matrigel-induced 3D reconstruction of eccrine sweat gland-like structures
EGFR inhibitor AG1478 blocks the formation of 3D structures mainly through ERK signaling pathway in Matrigel-induced 3D reconstruction of eccrine sweat gland-like structures
复制标题
EGFR抑制剂AG1478在Matrigel诱导的小汗腺样结构3D重建中主要通过ERK信号通路阻断3D结构的形成
DOI：
10.1007/s10735-020-09869-6
复制
发表时间：
2020-03-26
影响因子：
3.2
通讯作者：
Li, Haihong
中科院分区：
文献类型：
--
作者：
Chen, Liyun;Du, Lijie;Li, Haihong
EGFR信号在外泌汗腺的发育中起重要作用。我们以前证明，Matrigel诱导外泌汗腺细胞重建外泌汗腺的三维（3D）结构，但其机制仍然是未知的。在本研究中，外泌汗腺细胞在3D Matrigel中培养，并分别向培养基中加入EGFR抑制剂AG 1478或MEK 1/2抑制剂U 0126。观察外分泌汗腺样结构的三维重建形态，检测磷酸化EGFR的定位，并检测EGFR、磷酸化EGFR、磷酸化JAK、磷酸化AKT和磷酸化ERK的蛋白水平。结果显示，从3D培养物的第0天开始用AG 1478处理的细胞阻断了球状体样结构的形成。AG 1478给药导致磷酸化EGFR减少，伴随磷酸化ERK 1/2下调，但不导致磷酸化JAK或磷酸化AKT下调。U 0126处理后，磷酸化EGFR和磷酸化ERK减少，仅形成少量3D结构。我们的结论是，EGFR在基质胶诱导的外分泌汗腺样结构的三维结构中起着重要作用，ERK 1/2信号转导至少部分地负责EGFR的作用。
EGFR signaling plays important roles in the development of eccrine sweat glands. We previously demonstrate that Matrigel induces eccrine sweat gland cells to reconstruct the three-dimensional (3D) structures of eccrine sweat glands, but the mechanisms are still unknown. In the study, eccrine sweat gland cells were cultured within a 3D Matrigel, and EGFR inhibitor AG1478, or MEK1/2 inhibitor U0126, were added to the medium respectively. The morphology of the 3D-reconstructed eccrine sweat gland-like structures was observed, the localization of phospho-EGFR was detected, and protein levels of EGFR, phospho-EGFR, phospho-JAK, phospho-AKT and phospho-ERK were examined. The results showed that cells treatment with AG1478 from Day 0 of 3D cultures blocked formation of spheroid-like structures. AG1478 administration caused reduced phospho-EGFR, concomitant with downregulation of phospho-ERK1/2, but not phospho-JAK or phospho-AKT. Phospho-EGFR and phospho-ERK were reduced, and only a small number of 3D-structures were formed following treatment with U0126. We conclude that EGFR plays important roles in Matrigel-induced 3D structures of eccrine sweat gland-like structures, and ERK1/2 signaling is responsible, at least in part, for the effect of EGFR.