Rajabi Ali, Safaralizadeh Reza, Saber Ali, Pourmahdi Mahsa, Teimourian Shahram, Montazeri Vahid, Fakhrjou Ashraf, Hosseinpourfeizi Mohammadali
Department of Animal Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Iran.
Dr. Saber Medical Genetics Laboratory, Almas Complex, Namaz Blvd, Rasht, Gilan, Iran.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Feb;398(2):1815-1823. doi: 10.1007/s00210-024-03383-2. Epub 2024 Aug 23.
Breast cancer (BC) is the second most common malignancy worldwide. ADGRL4, as a modulator of angiogenesis, undergoes various epigenetic modifications affecting its biological functions. In this study, we aimed to assess ADGRL4 promoter methylation status and its expression levels in primary breast tumors and to evaluate its potency as a plausible prognostic biomarker in BC. Furthermore, we evaluated the effect of melatonin on ADGRL4 expression and viability of BC cells in vitro. One hundred breast tumor tissue samples and adjacent non-tumor tissues were collected, followed by DNA isolation, bisulfite conversion, qRT-PCR, qMSP assay, and immunoblotting. In addition, four BC cell lines were treated with melatonin and subjected to ADGRL4 expression analysis and apoptosis assay. We found a significant correlation between ADGRL4 expression levels and HER2 status and stage of disease (P < 0.05). We observed a substantial attenuation in ADGRL4 promoter methylation in tumor samples compared to marginal non-tumor samples. A significantly lower expression of ADGRL4 was detected in two BC cell lines in the presence of melatonin. MCF-7 and BT474 melatonin-treated cell lines showed a significantly higher number of apoptotic cells than non-treated cells (P < 0.0001). Based on the receiver operating characteristic (ROC) curve analysis, ADGRL4 expression and ADGRL4 promoter methylation status showed moderate prognostic value. We found that melatonin has anti-cancer effects on BC cells. In addition, ADGRL4 expression can potentially be used as a prognostic biomarker in BC.
乳腺癌（BC）是全球第二常见的恶性肿瘤。ADGRL4作为血管生成的调节因子，会经历各种表观遗传修饰，影响其生物学功能。在本研究中，我们旨在评估原发性乳腺肿瘤中ADGRL4启动子的甲基化状态及其表达水平，并评估其作为BC中一种合理的预后生物标志物的潜力。此外，我们在体外评估了褪黑素对BC细胞中ADGRL4表达和活力的影响。收集了100份乳腺肿瘤组织样本和相邻的非肿瘤组织，随后进行DNA提取、亚硫酸氢盐转化、qRT-PCR、qMSP分析和免疫印迹。此外，用褪黑素处理四种BC细胞系，并进行ADGRL4表达分析和凋亡检测。我们发现ADGRL4表达水平与HER2状态和疾病分期之间存在显著相关性（P < 0.05）。与边缘非肿瘤样本相比，我们观察到肿瘤样本中ADGRL4启动子甲基化显著减弱。在存在褪黑素的情况下，在两种BC细胞系中检测到ADGRL4表达显著降低。与未处理的细胞相比，MCF-7和BT474褪黑素处理的细胞系显示凋亡细胞数量显著增加（P < 0.0001）。基于受试者工作特征（ROC）曲线分析，ADGRL4表达和ADGRL4启动子甲基化状态显示出中等的预后价值。我们发现褪黑素对BC细胞具有抗癌作用。此外，ADGRL4表达有可能用作BC的预后生物标志物。
