Bandini Arianna, Banchi Marta, Orlandi Paola, Vaglini Francesca, Alì Greta, Fontanini Gabriella, Ottani Alessandra, Giuliani Daniela, Vandini Eleonora, Francia Giulio, Carli Marco, Scarselli Marco, Bocci Guido
Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Università di Pisa, 56126 Pisa, Italy.
Dipartimento di Medicina Clinica e Sperimentale, Università di Pisa, 56126 Pisa, Italy.
J Clin Med. 2025 Feb 11;14(4):1165. doi: 10.3390/jcm14041165.
MC4R expression and its role in colorectal and anaplastic thyroid cancers, where resistance to therapy and lack of standard treatments remain significant challenges, are poorly understood. This study aimed to investigate MC4R as a potential therapeutic target in these cancers using the selective antagonist ML00253764 (ML), alone and in combination with vinorelbine (VNR) and irinotecan (or its active metabolite SN-38). Human colorectal adenocarcinoma HT-29, Caco-2, and anaplastic thyroid carcinoma 8305C cell lines were used. MC4R expression was assessed by Real-Time PCR with validated primers (Assay ID Hs00271877_s1), immunofluorescence, and Western blotting. Proliferation and apoptosis assays were conducted with ML, and synergy with VNR and SN-38 was evaluated by Combination Index and Loewe methods. ERK1/2 phosphorylation was measured using an ELISA assay. In vivo studies were conducted by injecting tumor cells into Athymic Nude-Foxn1nu mice, treated with ML, VNR, irinotecan, or their combinations. MC4R expression was confirmed in all cell lines. ML treatment inhibited MC4R, producing antiproliferative and pro-apoptotic effects, with IC values of 7667 ± 2144.6 nM (8305C), 806.4 ± 321.8 nM (HT-29), and 2993 ± 1135.2 nM (Caco-2). In combination with VNR and SN-38, ML exhibited significant synergy in vitro and reduced tumor volume in vivo without causing weight loss or adverse effects in mice. This study identifies ML as a promising therapeutic agent that, when combined with chemotherapy, may offer a novel strategy for treating colorectal and anaplastic thyroid cancers.
黑素皮质素4受体（MC4R）的表达及其在结直肠癌和间变性甲状腺癌中的作用尚不清楚，在这些癌症中，对治疗的耐药性和缺乏标准治疗方法仍然是重大挑战。本研究旨在研究MC4R作为这些癌症潜在治疗靶点的作用，单独使用选择性拮抗剂ML00253764（ML），以及与长春瑞滨（VNR）和伊立替康（或其活性代谢物SN-38）联合使用。使用了人结肠腺癌HT-29、Caco-2和间变性甲状腺癌8305C细胞系。通过使用经过验证的引物（检测ID Hs00271877_s1）的实时PCR、免疫荧光和蛋白质印迹法评估MC4R的表达。用ML进行增殖和凋亡检测，并通过联合指数和洛伊方法评估与VNR和SN-38的协同作用。使用酶联免疫吸附测定法测量细胞外信号调节激酶1/2（ERK1/2）的磷酸化。通过将肿瘤细胞注射到无胸腺裸鼠（Foxn1nu）中进行体内研究，用ML、VNR、伊立替康或它们的组合进行治疗。在所有细胞系中均证实了MC4R的表达。ML治疗可抑制MC4R，产生抗增殖和促凋亡作用，8305C细胞系的半数抑制浓度（IC）值为7667±2144.6 nM，HT-29细胞系为806.4±321.8 nM，Caco-2细胞系为2993±1135.2 nM。与VNR和SN-38联合使用时，ML在体外表现出显著的协同作用，在体内可减小肿瘤体积，且不会导致小鼠体重减轻或产生不良反应。本研究确定ML是一种有前景的治疗药物，与化疗联合使用时，可能为治疗结直肠癌和间变性甲状腺癌提供一种新策略。
