Nikolic Ivana, Lazovic Aleksandar, Stanisavljevic Isidora, Andjelkovic Marija, Popovic Suzana, Pavlovic Sladjana, Jurisevic Milena, Mitrovic Marina
Department of Medical Biochemistry, Faculty of Medical Sciences, University of Kragujevac, Svetozara Markovića, 69, 34000 Kragujevac, Serbia.
Department of Surgery, Faculty of Medical Sciences, University of Kragujevac, Svetozar Markovic 69, 34000 Kragujevac, Serbia.
Eur J Pharm Sci. 2025 Sep 1;212:107165. doi: 10.1016/j.ejps.2025.107165. Epub 2025 Jun 10.
Cancer remains the second leading cause of mortality worldwide, underscoring the urgent need for novel therapeutic strategies. Drug repurposing is an effective strategy to address current cancer challenges, such as the resistance and toxicity associated with traditional chemotherapy. Among the various psychotropic drugs, antidepressants are emerging as promising candidates due to their demonstrated anticancer activity.
We evaluated duloxetine (DUL), a serotonin-norepinephrine reuptake inhibitor, for anticancer and immunomodulatory activity by performing comprehensive in vitro and in vivo experiments. In HCT116 (colon), HeLa (cervical), and MDA-MB-231 (triple-negative breast cancer) human cancer cells, and 4T1 mouse breast cancer cells, we investigated DUL's cytotoxicity, apoptosis, autophagy, cell cycle arrest, and migration inhibition through MTT assay, flow cytometry, and immunofluorescence methods. An orthotopic mouse model of breast cancer was utilized to investigate the in vivo tumor inhibitory effects of DUL, along with its systemic toxicity and immunomodulatory properties. Gene expression (p53, Beclin-1), cytokine profile, and CD3T cell activation were examined by mRNA sequencing and ELISA kits to explore underlying mechanisms.
For the first time, our in vitro results indicated that DUL caused dose-dependent cytotoxicity, apoptosis, autophagy, and cell cycle arrest in all cancer lines tested, which was more selective than 5-fluorouracil (5-FU). Mechanistically, DUL modulated apoptotic (Bcl-2, Bax, caspase-3), autophagic (p62), and survival (pAkt) signaling, disrupted mitochondrial membrane potential, and inhibited cell migration. In vivo, DUL inhibited tumor growth without inducing hepatic or renal toxicity. Notably, DUL stimulated the production of pro-inflammatory cytokines (IFN-γ, IL-1β, TNF-α), enhanced the immunity of CD3T cells, and increased the expression of pro-apoptotic p53 and autophagic Beclin-1 genes, reflecting its dual antitumor and immunomodulatory actions.
Our findings revealed for the first time that DUL is a promising repurposed drug for cancer treatment, as it has demonstrated proven antitumor efficacy and immune-stimulating properties. This novel dual role of DUL warrants further investigation in clinical oncology.
癌症仍然是全球第二大死因，这凸显了对新型治疗策略的迫切需求。药物重新利用是应对当前癌症挑战（如与传统化疗相关的耐药性和毒性）的有效策略。在各种精神药物中，抗抑郁药因其已证明的抗癌活性而成为有前景的候选药物。
我们通过进行全面的体外和体内实验，评估了5-羟色胺-去甲肾上腺素再摄取抑制剂度洛西汀（DUL）的抗癌和免疫调节活性。在人结肠癌细胞HCT116、宫颈癌细胞HeLa、三阴性乳腺癌细胞MDA-MB-231以及小鼠乳腺癌细胞4T1中，我们通过MTT法、流式细胞术和免疫荧光法研究了DUL的细胞毒性、凋亡、自噬、细胞周期阻滞和迁移抑制作用。利用原位小鼠乳腺癌模型研究了DUL的体内肿瘤抑制作用及其全身毒性和免疫调节特性。通过mRNA测序和酶联免疫吸附测定试剂盒检测基因表达（p53、Beclin-1）、细胞因子谱和CD3T细胞活化情况，以探索潜在机制。
我们的体外研究结果首次表明，DUL在所有测试的癌细胞系中均引起剂量依赖性细胞毒性、凋亡、自噬和细胞周期阻滞，其选择性高于5-氟尿嘧啶（5-FU）。从机制上讲，DUL调节凋亡信号（Bcl-2、Bax、caspase-3）、自噬信号（p62）和生存信号（pAkt），破坏线粒体膜电位，并抑制细胞迁移。在体内，DUL抑制肿瘤生长而不诱导肝毒性或肾毒性。值得注意的是，DUL刺激促炎细胞因子（IFN-γ、IL-1β、TNF-α）的产生，增强CD3T细胞的免疫力，并增加促凋亡p53和自噬相关Beclin-1基因的表达，反映了其双重抗肿瘤和免疫调节作用。
我们的研究结果首次表明，DUL是一种有前景的癌症治疗重新利用药物，因为它已证明具有抗肿瘤疗效和免疫刺激特性。DUL的这种新型双重作用值得在临床肿瘤学中进一步研究。
