Alper Meltem, Sav Feyza Nur, Keleş Yasemin, Eroğlu Kübra Paspal, Keskin Saliha Derya, Köçkar Feray
Department of Translational Oncology, Institute of Oncology, Dokuz Eylül University, Izmir, Turkey.
Department of Molecular Biology and Genetics, Faculty of Science and Literature, Balıkesir University, Balikesir, Turkey.
Mol Biol Rep. 2025 Feb 19;52(1):246. doi: 10.1007/s11033-025-10342-4.
ADAMTSs are extracellular matrix metalloproteinases that mainly process extracellular matrix components and closely related tumorigenesis. ADAMTS-8 is an anti-angiogenic member of the family and is dysregulated in common cancers. The tumor suppressor function of the ADAMTS-8 has been demonstrated in colorectal cancer. Although ADAMTS-8 plays a critical role in tumor progression, transcriptional regulatory features haven't been studied yet.
The human ADAMTS-8 promoter was cloned into the pMetLuc Reporter vector. Basal promoter activity and the effect of the IL-6 on ADAMTS-8 promoter activity were determined by transient transfection assays in SW480 cells. QRT-PCR and Western blot analyses assessed the impact of IL-6 on ADAMTS-8 mRNA and protein expressions. Functional binding of the specific transcription factors to the ADAMTS-8 promoter region was evaluated by ChIP qPCR and EMSA.
Our results demonstrated that the ADAMTS-8 promoter includes multiple binding sites for transcription factors that could be activated in the inflammatory pathways. IL-6 stimulation increased ADAMTS-8 promoter activity, also mRNA, and protein expressions. Pathway inhibition studies showed that IL-6-mediated induction of ADAMTS-8 was achieved through p38/MAPK, NF-κB, PI3K, and SAPK/JNK pathways. STATs, Elk-1, and c-Jun functionally bind to the ADAMTS-8 promoter region.
It can be concluded that inflammation is a strong positive regulator of the ADAMTS-8 gene.
含血小板解聚蛋白基序的金属蛋白酶（ADAMTSs）是细胞外基质金属蛋白酶，主要作用于细胞外基质成分，与肿瘤发生密切相关。ADAMTS-8是该家族的一种抗血管生成成员，在常见癌症中表达失调。ADAMTS-8的肿瘤抑制功能已在结直肠癌中得到证实。尽管ADAMTS-8在肿瘤进展中起关键作用，但其转录调控特征尚未得到研究。
将人ADAMTS-8启动子克隆到pMetLuc报告载体中。通过在SW480细胞中进行瞬时转染试验，测定基础启动子活性以及白细胞介素-6（IL-6）对ADAMTS-8启动子活性的影响。实时定量聚合酶链反应（QRT-PCR）和蛋白质免疫印迹分析评估IL-6对ADAMTS-8信使核糖核酸（mRNA）和蛋白质表达的影响。通过染色质免疫沉淀定量聚合酶链反应（ChIP qPCR）和电泳迁移率变动分析（EMSA）评估特定转录因子与ADAMTS-8启动子区域的功能性结合。
我们的结果表明，ADAMTS-8启动子包含多个可在炎症途径中被激活的转录因子结合位点。IL-6刺激增加了ADAMTS-8启动子活性、mRNA和蛋白质表达。通路抑制研究表明，IL-6介导的ADAMTS-8诱导是通过p38/丝裂原活化蛋白激酶（MAPK）、核因子κB（NF-κB）、磷脂酰肌醇-3激酶（PI3K）和应激激活蛋白激酶/应激活化蛋白激酶（SAPK/JNK）通路实现的。信号转导和转录激活因子（STATs）、 Elk-1和c-Jun在功能上与ADAMTS-8启动子区域结合。
可以得出结论，炎症是ADAMTS-8基因的强正调控因子。
