Department of Pathology, Peking Union Medical College Hospital, Research Center for Molecular Pathology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, P.R. China.
Oncol Rep. 2020 May;43(5):1558-1568. doi: 10.3892/or.2020.7523. Epub 2020 Feb 27.
Amphiregulin (AREG) is a member of the epidermal growth factor (EGF) family and is expressed in a plethora of cancers. The biological roles of AREG in the regulation of the epithelial‑mesenchymal transition (EMT) in pancreatic cancer remain unclear. To investigate the expression of epidermal growth factor receptor (EGFR) and AREG in pancreatic cancer cell lines, RT‑qPCR, western blot analysis, and ELISA were performed. RNAi and exogenous AREG treatment were used to alter AREG expression. Wound‑healing and Transwell assays were performed to evaluate cell migration and invasion abilities. Western blot analysis and immunofluorescence staining were utilized to detect the expression of EMT markers. The protein expression of potential key factors involved in EMT, as well as those of the ERK, AKT, STAT3 and NF‑κB pathways, were analysed by western blotting. The role of AREG in tumour growth in vivo was further determined using an orthotopic model of pancreatic cancer. Knockdown of AREG inhibited AsPC‑1 cell migration and invasion. AREG knockdown upregulated E‑cadherin but downregulated vimentin, Snail and Slug expression in AsPC‑1 cells. In addition, AREG stimulation increased cell migration, invasion and EMT in PANC‑1 cells, and an NF‑κB inhibitor decreased AREG‑induced cell migration, invasion and EMT in PANC‑1 cells. AREG stimulation increased the nuclear accumulation of NF‑κB through the EGFR/ERK signalling pathway to induce EMT. Tumour growth and metastasis were decreased by AREG silencing in an orthotopic model of pancreatic cancer. AREG may play a critical role in cell migration, invasion, and EMT by activating the EGFR/ERK/NF‑κB signalling pathway in pancreatic cancer cells.
双调蛋白 (AREG) 是表皮生长因子 (EGF) 家族的一员，在多种癌症中表达。AREG 在胰腺癌上皮-间质转化 (EMT) 中的调控作用尚不清楚。为了研究表皮生长因子受体 (EGFR) 和 AREG 在胰腺癌细胞系中的表达，进行了 RT-qPCR、western blot 分析和 ELISA。使用 RNAi 和外源性 AREG 处理来改变 AREG 表达。进行划痕愈合和 Transwell 测定以评估细胞迁移和侵袭能力。使用 western blot 分析和免疫荧光染色来检测 EMT 标志物的表达。通过 western blot 分析来分析 EMT 涉及的潜在关键因子以及 ERK、AKT、STAT3 和 NF-κB 通路的蛋白表达。进一步使用胰腺癌的原位模型来确定 AREG 在肿瘤生长中的作用。AREG 敲低抑制了 AsPC-1 细胞的迁移和侵袭。AREG 敲低在上皮细胞中上调了 E-钙黏蛋白但下调了波形蛋白、Snail 和 Slug 的表达。此外，AREG 刺激增加了 PANC-1 细胞的细胞迁移、侵袭和 EMT，NF-κB 抑制剂降低了 PANC-1 细胞中 AREG 诱导的细胞迁移、侵袭和 EMT。AREG 刺激通过 EGFR/ERK 信号通路增加 NF-κB 的核积累，从而诱导 EMT。在胰腺癌的原位模型中，AREG 沉默降低了肿瘤生长和转移。AREG 可能通过激活胰腺癌细胞中的 EGFR/ERK/NF-κB 信号通路在细胞迁移、侵袭和 EMT 中发挥关键作用。
