Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand.
Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand.
Arch Oral Biol. 2022 Feb;134:105335. doi: 10.1016/j.archoralbio.2021.105335. Epub 2021 Dec 3.
To investigate the in vitro effects of inhibiting galectin-1 using the small-molecule inhibitor OTX008 on oral squamous cell carcinoma (OSCC) cell lines and the role of the MAPK pathway.
One normal oral keratinocyte (NOK) and three OSCC cell lines were cultured in vitro and the expression of galectin-1 protein by each quantified using ELISA. Cell lines were treated with galectin-1 (50, 100 and 150 ng/mL) or OTX008 (12.5, 25, 50 and 100 μg/mL) and cell viability assayed (n = 3). OSCC cell lines with and without 25 μg/mL OTX008 (n = 3) treatment for 48 h, were analysed using qRT-PCR with a custom array, to assess relative gene expression.
All cell lines were found to express galectin-1 protein. Exogenous galectin-1 significantly reduced cell viability in one OSCC cell line over time while the others were only minimally affected. OTX008 treatment reduced cell viability in a dose and time-dependent manner in all cell lines and this was associated with significant regulation of FOS gene expression in the OSCC cell lines.
OTX008 decreased the viability of OSCC and NOK cells in a dose-dependent manner. The significant regulation of FOS suggests OTX008 causes early induction of the MAPK pathway via the immediate response gene FOS as a subunit of the AP-1 complex.
研究小分子抑制剂 OTX008 抑制半乳糖凝集素-1（galectin-1）对口腔鳞状细胞癌（OSCC）细胞系的体外作用及其对 MAPK 通路的影响。
体外培养 1 株正常口腔角质细胞（NOK）和 3 株 OSCC 细胞系，采用 ELISA 法检测 galectin-1 蛋白的表达。用 galectin-1（50、100 和 150ng/mL）或 OTX008（12.5、25、50 和 100μg/mL）处理细胞系，检测细胞活力（n=3）。用 25μg/mL OTX008 处理 48h 后，采用 qRT-PCR 检测经定制阵列处理的 OSCC 细胞系的相对基因表达。
所有细胞系均表达 galectin-1 蛋白。外源性 galectin-1 随时间推移明显降低了一种 OSCC 细胞系的细胞活力，而其他细胞系仅受到轻微影响。OTX008 处理以剂量和时间依赖的方式降低了所有细胞系的细胞活力，并且与 OSCC 细胞系中 FOS 基因表达的显著调节相关。
OTX008 以剂量依赖的方式降低了 OSCC 和 NOK 细胞的活力。FOS 的显著调节表明，OTX008 通过立即反应基因 FOS 作为 AP-1 复合物的亚基，早期诱导 MAPK 通路。
