Chen Wanling, Ye Sai, Wang Xiaoning, Qian Jiajun, Xia Liang, Tian Zhen
Department of Oral Pathology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
College of Stomatology, Shanghai Jiao Tong University, Shanghai, China.
J Oral Pathol Med. 2023 Jan;52(1):63-71. doi: 10.1111/jop.13388. Epub 2022 Dec 5.
Soluble E-cadherin (sEcad), a tumor suppressor gene, has pro-oncogenic effects by binding to human epithelial growth factor receptor 2 (HER-2). In our previous study, 1/3 of carcinoma ex pleomorphic adenoma (CXPA) cases had HER-2 amplification, which is associated with tumorigenesis and malignancy. This study examines the role of sEcad in HER-2 amplified CXPA.
Immunohistochemistry was used to examine E-cadherin (Ecad) expression in HER-2-amplified CXPA samples (n = 35). Western blot and ELISA were used to detect sEcad in two samples with Ecad and HER-2 overexpression and CXPA cell line. Lentivirus-mediated transfection was performed to knock down sEcad in CXPA cells. The cell proliferation, wound healing, and transwell assays were used to compare sEcad-knockdown cells with cells pretreated with recombinant human sEcad (rhEcad/Fc). sEcad and HER-2 interaction was determined through co-immunoprecipitation. RNA-sequencing, differential expression analysis, GO and KEGG analysis were used to identify sEcad-related signaling pathways and their protein phosphorylation levels were verified by western blotting.
Ecad was overexpressed in 77.1% of HER-2-positive CXPA, and sEcad was found in the CXPA cell line and two samples. sEcad promoted CXPA migration and invasion in vitro without sEcad and HER-2 interaction. sEcad-related differentially expressed genes were enriched in the IL-17, cAMP, and MAPK signaling pathways. Furthermore, sEcad activated the phosphorylation of Akt and MAPK/ERK signaling pathways.
Most HER-2 CXPAs express Ecad. sEcad could affect the invasiveness and migration of in vitro CXPA cells without HER-2. sEcad may be a therapeutic biomarker for CXPA patients.
可溶性E-钙黏蛋白（sEcad）作为一种肿瘤抑制基因，通过与人类表皮生长因子受体2（HER-2）结合发挥促癌作用。在我们之前的研究中，三分之一的多形性腺瘤癌变（CXPA）病例存在HER-2扩增，这与肿瘤发生和恶性程度相关。本研究旨在探讨sEcad在HER-2扩增的CXPA中的作用。
采用免疫组织化学法检测HER-2扩增的CXPA样本（n = 35）中E-钙黏蛋白（Ecad）的表达。利用蛋白质免疫印迹法和酶联免疫吸附测定法检测两个Ecad和HER-2过表达样本及CXPA细胞系中的sEcad。通过慢病毒介导的转染敲低CXPA细胞中的sEcad。采用细胞增殖、伤口愈合和Transwell实验，将sEcad敲低的细胞与经重组人sEcad（rhEcad/Fc）预处理的细胞进行比较。通过免疫共沉淀确定sEcad与HER-2的相互作用。利用RNA测序、差异表达分析、基因本体（GO）和京都基因与基因组百科全书（KEGG）分析来鉴定与sEcad相关的信号通路，并通过蛋白质免疫印迹法验证其蛋白质磷酸化水平。
77.1%的HER-2阳性CXPA中Ecad过表达，在CXPA细胞系和两个样本中发现了sEcad。sEcad在体外促进CXPA迁移和侵袭，且不存在sEcad与HER-2的相互作用。与sEcad相关的差异表达基因在白细胞介素-17、环磷酸腺苷（cAMP）和丝裂原活化蛋白激酶（MAPK）信号通路中富集。此外，sEcad激活了蛋白激酶B（Akt）和MAPK/细胞外信号调节激酶（ERK）信号通路的磷酸化。
大多数HER-2阳性的CXPA表达Ecad。sEcad可在无HER-2的情况下影响体外CXPA细胞的侵袭性和迁移能力。sEcad可能是CXPA患者的一种治疗生物标志物。
Zotero 插件