Pérez-de-Oliveira Maria Eduarda, Wagner Vivian Petersen, Bingle Colin D, Vargas Pablo Agustin, Bingle Lynne
Department of Oral Diagnosis, Piracicaba Dental School, Universidade Estadual de Campinas, Piracicaba, São Paulo, Brazil; School of Clinical Dentistry, University of Sheffield, Sheffield, United Kingdom.
Department of Oral Diagnosis, Piracicaba Dental School, Universidade Estadual de Campinas, Piracicaba, São Paulo, Brazil; School of Clinical Dentistry, University of Sheffield, Sheffield, United Kingdom; Department of Oral Medicine, School of Dentistry, Universidade de São Paulo, São Paulo, São Paulo, Brazil.
Oral Oncol. 2024 Dec;159:107029. doi: 10.1016/j.oraloncology.2024.107029. Epub 2024 Sep 26.
Mucoepidermoid carcinoma (MEC) is the most common malignant salivary gland tumour with around 50 % of cases carrying the CRTC1-MAML2 translocation. The CREB pathway has been associated with the transforming activity of this translocation. The aim of this study was to determine the effects of CREB inhibition on MEC cell behaviour in vitro.
Two translocation-positive (UM-HMC-2 and H292) and one translocation-negative (H253) MEC cell lines were treated with 666.15, a CREB inhibitor. Drug IC50 doses were determined for each cell line. Clonogenic and spheroid assays were used to assess survival, including percentage of cancer stem cells, and transwell and scratch assays evaluated invasive and migratory capacities, respectively. Immunofluorescence staining was used to determine E-cadherin expression.
CREB inhibition significantly reduced the number of surviving colonies and spheroids and delayed cell invasion in all cell lines, but this was more significant in the fusion positive, UM-HMC-2 cells. The expression of E-cadherin was significantly higher in treated UM-HMC-2 and H292 cells.
CREB inhibition with 666.15 impaired key MEC oncogenic behaviours associated with metastasis and drug resistance, including cell invasion and survival.
黏液表皮样癌（MEC）是最常见的涎腺恶性肿瘤，约50%的病例存在CRTC1-MAML2易位。CREB信号通路与这种易位的转化活性有关。本研究的目的是确定抑制CREB对MEC细胞体外行为的影响。
用CREB抑制剂666.15处理两株易位阳性（UM-HMC-2和H292）和一株易位阴性（H253）的MEC细胞系。测定每种细胞系的药物IC50剂量。采用克隆形成试验和球体形成试验评估细胞存活情况，包括癌干细胞百分比，采用Transwell试验和划痕试验分别评估细胞的侵袭能力和迁移能力。采用免疫荧光染色法检测E-钙黏蛋白的表达。
抑制CREB可显著减少所有细胞系中存活的集落和球体数量，并延迟细胞侵袭，但在融合阳性的UM-HMC-2细胞中更为显著。在经处理的UM-HMC-2和H292细胞中，E-钙黏蛋白的表达显著升高。
用666.15抑制CREB可损害与转移和耐药相关的关键MEC致癌行为，包括细胞侵袭和存活。
