Zhang Meng-Jie, Wu Cong-Cong, Wang Shuo, Yang Lei-Lei, Sun Zhi-Jun
The State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Department of Oral and Maxillofacial Head Neck Oncology, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
Oral Dis. 2023 Jan;29(1):175-187. doi: 10.1111/odi.14045. Epub 2021 Oct 21.
Adenoid cystic carcinoma (AdCC) and mucoepidermoid carcinoma (MEC) are the two most frequent malignancies of salivary glands. This study aims to explore the expression and migration of LAG3, TIM3, and A2aR in AdCC and MEC, and the potential relationship with oncogenic signaling molecules and immunosuppressive cytokines.
Custom made human salivary gland tissue microarrays included 81 AdCCs, 52 MECs, 76 normal salivary glands (NSG), and 14 pleomorphic adenoma (PMA) samples. Immunohistochemical analysis of lymphocyte activation gene 3 (LAG3), T-cell immunoglobulin and mucin domain-containing protein 3 (TIM3), adenosine 2a receptor (A2aR), oncogenic phosphorylated S6 kinase (p-S6) and ERK (p-ERK ), and TGF-β1 was performed with salivary gland tissue microarrays of human samples. The correlation of the immunostaining was analyzed based on a digital pathological system, and data were evaluated by hierarchical cluster. Further in vitro studies of knockdown immune checkpoints LAG3, TIM3, and A2aR were carried out by siRNA transfection.
The expression levels of LAG3, TIM3, and A2aR were remarkably increased in AdCC and MEC, compared with NSG and PMA samples, but were independent of pathology grade. They were closely correlated with TGF-β1, slightly related to p-ERK and p-S6. After the knockdown of immune checkpoints LAG3, TIM3, and A2aR, the migration of SACC-LM cell line was significantly reduced.
These results suggested that LAG3, TIM3, and A2aR are overexpressed in AdCC and MEC, may promote migration of SACC-LM cell and correlated with TGF-β1 and oncogenic signaling pathways.
腺样囊性癌（AdCC）和黏液表皮样癌（MEC）是唾液腺最常见的两种恶性肿瘤。本研究旨在探讨淋巴细胞激活基因3（LAG3）、T细胞免疫球蛋白黏蛋白结构域3（TIM3）和腺苷2A受体（A2aR）在AdCC和MEC中的表达及迁移情况，以及与致癌信号分子和免疫抑制细胞因子的潜在关系。
定制的人唾液腺组织芯片包含81例AdCC、52例MEC、76例正常唾液腺（NSG）和14例多形性腺瘤（PMA）样本。采用人唾液腺组织芯片对淋巴细胞激活基因3（LAG3）、T细胞免疫球蛋白和黏蛋白结构域含蛋白3（TIM3）、腺苷2A受体（A2aR）、致癌性磷酸化S6激酶（p-S6）和细胞外信号调节激酶（p-ERK）以及转化生长因子-β1（TGF-β1）进行免疫组化分析。基于数字病理系统分析免疫染色的相关性，并通过层次聚类评估数据。通过小干扰RNA转染对免疫检查点LAG3、TIM3和A2aR进行进一步的体外敲低研究。
与NSG和PMA样本相比，AdCC和MEC中LAG3、TIM3和A2aR的表达水平显著升高，但与病理分级无关。它们与TGF-β1密切相关，与p-ERK和p-S6轻度相关。免疫检查点LAG3、TIM3和A2aR敲低后，SACC-LM细胞系的迁移显著减少。
这些结果表明，LAG3、TIM3和A2aR在AdCC和MEC中过表达，可能促进SACC-LM细胞迁移，并与TGF-β1和致癌信号通路相关。
