Cao Yuhui, Zhou Zhihui, He Shuai, Liu Wenhui
Department of Stomatology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325088, China.
Department of Stomatology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325088, China.
Arch Oral Biol. 2024 Sep;165:106028. doi: 10.1016/j.archoralbio.2024.106028. Epub 2024 Jun 15.
This study was designed to investigate the biological role and the reaction mechanism of Tweety family member 3 (TTYH3) in oral squamous cell carcinoma (OSCC).
The mRNA and protein expressions of TTYH3 were assessed with RT-qPCR and western blot. After silencing TTYH3 expression, the proliferation of OSCC cells were detected using cell counting kit-8 (CCK-8) assay, 5-ethynyl-2'-deoxyuridine (EdU) staining and colony formation assay. Cell migration and invasion were detected using wound healing and transwell. Gelatin zymography protease assay was used to detect matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-2 (MMP9) activity and western blot was used to detect the expressions of proteins associated with proliferation and epithelial-mesenchymal transition (EMT). The mRNA expression of TTYH3 in THP-1-derived macrophage was detected using real-time reverse transcriptase-polymerase chain reaction (RT-qPCR). The number of CD86-positive cells and CD206-positive cells was detected using immunofluorescence assay. RT-qPCR was used to detect the expressions of M2 markers arginase 1 (ARG1), chitinase-like 3 (YM1) and mannose receptor C-type 1 (MRC1).
In this study, it was found that TTYH3 expression was upregulated in OSCC cell lines and TTYH3 knockdown could inhibit the proliferation, migration, invasion and EMT process in OSCC via suppressing M2 polarization of tumor-associated macrophages.
Collectively, TTYH3 facilitated the progression of OSCC through the regulation of tumor-associated macrophages polarization.
本研究旨在探讨Tweety家族成员3（TTYH3）在口腔鳞状细胞癌（OSCC）中的生物学作用及反应机制。
采用RT-qPCR和蛋白质免疫印迹法评估TTYH3的mRNA和蛋白表达。沉默TTYH3表达后，使用细胞计数试剂盒-8（CCK-8）检测、5-乙炔基-2'-脱氧尿苷（EdU）染色和集落形成试验检测OSCC细胞的增殖。采用伤口愈合试验和Transwell检测细胞迁移和侵袭。使用明胶酶谱蛋白酶试验检测基质金属蛋白酶-2（MMP2）和基质金属蛋白酶-9（MMP9）活性，并用蛋白质免疫印迹法检测与增殖和上皮-间质转化（EMT）相关的蛋白表达。采用实时逆转录-聚合酶链反应（RT-qPCR）检测THP-1来源巨噬细胞中TTYH3的mRNA表达。采用免疫荧光试验检测CD86阳性细胞和CD206阳性细胞的数量。采用RT-qPCR检测M2标志物精氨酸酶1（ARG1）、几丁质酶样3（YM1）和甘露糖受体C型1（MRC1）的表达。
本研究发现，TTYH3在OSCC细胞系中表达上调，敲低TTYH3可通过抑制肿瘤相关巨噬细胞的M2极化来抑制OSCC的增殖、迁移、侵袭和EMT过程。
综上所述，TTYH3通过调节肿瘤相关巨噬细胞极化促进OSCC进展。
