Squitti Rosanna, De Luca Anastasia, Severino Altea, Rizzo Gianluca, Marzi Federica, Amodio Luca Emanuele, Vicano Gabriella, Focaccio Antonio, Tondolo Vincenzo, Rongioletti Mauro
Department of Laboratory Science, Research and Development Division, Ospedale Isola Tiberina-Gemelli Isola, 00186 Rome, Italy.
Department of Theoretical and Applied Sciences, eCampus University, Viale Massenzio Masia 26, Novedrate, 22100 Como, Italy.
Int J Mol Sci. 2025 May 28;26(11):5163. doi: 10.3390/ijms26115163.
Sex steroid hormones and systemic iron metabolism are emerging as modulators of colorectal cancer (CRC) development and progression. However, information linking systemic factors to tumor characteristics and epithelial-mesenchymal transition (EMT) is limited, particularly in a sex-specific context. We measured serum levels of sex hormones [testosterone, estradiol, progesterone, Luteinizing Hormone (LH), Follicle-Stimulating Hormone (FSH), Carcinoembryonic antigen (CEA)] and iron-related biomarkers (iron, transferrin, ferritin, % transferrin saturation, ceruloplasmin, and the ceruloplasmin/transferrin ratio) in 82 CRC patients and 31 healthy controls. EMT-related proteins [mediator of ErbB2-driven cell motility 1 (MEMO1), E-cadherin, fibronectin, vimentin, and vinculin] were quantified by Western blotting in tumor and adjacent normal mucosa. Non-parametric tests and Spearman correlations were applied, stratified by sex and corrected for age and anemia where appropriate. Progesterone levels were significantly lower in male CRC patients (median 0.17 ng/mL vs. 0.20 ng/mL, = 0.04) and higher in female patients (0.17 ng/mL vs. 0.10 ng/mL, = 0.0077) compared with controls. The iron-related biomarkers indicated a pattern of iron deficiency, including in non-anemic patients, with reduced % transferrin saturation ( < 0.01) and an elevated ceruloplasmin/transferrin ratio ( = 0.02). Correlations were found between iron status, tumor stage, and hormonal levels. Progesterone correlated with EMT protein expression in healthy mucosa (e.g., fibronectin in females: ρ = 0.567, = 0.014; vimentin in males: ρ = -0.446, = 0.007), but not in tumor tissue. In the healthy mucosa of male patients, ceruloplasmin/transferrin correlated with MEMO1 (ρ = 0.419, = 0.04), vinculin (ρ = 0.299, = 0.041), and vimentin (ρ = 0.394, = 0.07); transferrin levels inversely correlated with MEMO1 expression (ρ = -0.392, = 0.032), and vimentin showed a positive correlation with serum iron (ρ = 0.350, = 0.043). Furthermore, fibronectin expression inversely correlated with iron in the sole tumor tissue of female patients (ρ = -0.366, = 0.040). These findings support the role of sex hormones and iron metabolism in CRC biology, suggesting that EMT might be accompanied by altered iron uptake and redox remodeling, which can enhance cellular motility and the metastatic potential.
性类固醇激素和全身铁代谢正逐渐成为结直肠癌（CRC）发生发展的调节因子。然而，将全身因素与肿瘤特征及上皮-间质转化（EMT）联系起来的信息有限，尤其是在性别特异性背景下。我们测定了82例CRC患者和31名健康对照者血清中的性激素水平[睾酮、雌二醇、孕酮、促黄体生成素（LH）、促卵泡生成素（FSH）、癌胚抗原（CEA）]以及铁相关生物标志物（铁、转铁蛋白、铁蛋白、转铁蛋白饱和度百分比、铜蓝蛋白以及铜蓝蛋白/转铁蛋白比值）。通过蛋白质免疫印迹法对肿瘤及相邻正常黏膜中的EMT相关蛋白[ErbB2驱动的细胞运动介质1（MEMO1）、E-钙黏蛋白、纤连蛋白、波形蛋白和纽蛋白]进行定量分析。采用非参数检验和Spearman相关性分析，按性别分层，并在适当情况下对年龄和贫血进行校正。与对照组相比，男性CRC患者的孕酮水平显著降低（中位数0.17 ng/mL对0.20 ng/mL，P = 0.04），而女性患者的孕酮水平较高（0.17 ng/mL对0.10 ng/mL，P = 0.0077）。铁相关生物标志物显示出缺铁模式，包括非贫血患者，转铁蛋白饱和度百分比降低（P < 0.01）且铜蓝蛋白/转铁蛋白比值升高（P = 0.02）。发现铁状态、肿瘤分期和激素水平之间存在相关性。孕酮与健康黏膜中的EMT蛋白表达相关（例如，女性中的纤连蛋白：ρ = 0.567，P = 0.014；男性中的波形蛋白：ρ = -0.446，P = 0.007），但与肿瘤组织无关。在男性患者的健康黏膜中，铜蓝蛋白/转铁蛋白与MEMO1相关（ρ = 0.419，P = 0.04）、纽蛋白（ρ = 0.299，P = 0.041）和波形蛋白（ρ = 0.394，P = 0.07）；转铁蛋白水平与MEMO1表达呈负相关（ρ = -0.392，P = 0.032），且波形蛋白与血清铁呈正相关（ρ = 0.350，P = 0.043）。此外，在女性患者的肿瘤组织中，纤连蛋白表达与铁呈负相关（ρ = -0.366，P = 0.040）。这些发现支持了性激素和铁代谢在CRC生物学中的作用，表明EMT可能伴随着铁摄取和氧化还原重塑的改变，这可增强细胞运动性和转移潜能。
