Işlek Köklü Zeynep, Şanverdi Elif Lidya, Karadağ Başak, Üçişik Mehmet Hikmet, Taşkan Ezgi, Şahin Fikrettin
Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, İstanbul, Turkiye.
Turk J Biol. 2024 Oct 14;48(6):401-413. doi: 10.55730/1300-0152.2715. eCollection 2024.
BACKGROUND/AIM: Melanoma arises from the uncontrolled multiplication of melanocytes, and poses an escalating global health concern. Despite the importance of early detection and surgical removal for effective treatment, metastatic melanoma poses treatment challenges, with limited options. Among optional therapies, including chemotherapy and immunotherapy, all-trans retinoic acid (ATRA), a natural metabolite of vitamin A, has shown promise in treating melanoma by inducing differentiation, apoptosis, growth arrest, and immune modulation in melanoma cells. However, ATRA treatment alone can lead to resistance and relapse. Furthermore, sphingomyelin (SM) was implicated in the inhibition of cell proliferation, differentiation, and apoptotic cell death during melanoma progression.
The combinational anticancer effects of ATRA and SM on an in vitro B16F10 melanoma model were investigated based on cell viability, apoptotic cell death, cell cycle progression, and gene expression levels; whereas the safety properties of the treatments were tested on RAW264.7 macrophages.
The combination of 123 μM of ATRA + 136 μM of SM was the most effective treatment, showing a 50% reduction in cell proliferation, leading to 53.91% apoptotic cell death in 48 h, and G2/M phase-cell cycle arrest in the B16F10 cells. While 123 μM of ATRA alone did not change the caspase 3 and Bax gene expressions, the combinational ATRA + SM treatment resulted in 2- and 5-fold increases in the gene expression level, respectively. A 13-fold increase in cyclin-dependent kinase inhibitor 2A was observed with the combinational ATRA + SM treatment, while suppressing the programmed death ligand 1 (PD-L1) expression by 0.5-fold.
Combinational ATRA and SM therapy could be a promising therapeutic approach for melanoma, potentially improving efficacy, while reducing toxicity to healthy cells.
背景/目的：黑色素瘤起源于黑素细胞的失控增殖，对全球健康构成日益严重的威胁。尽管早期检测和手术切除对于有效治疗至关重要，但转移性黑色素瘤的治疗仍面临挑战，治疗选择有限。在包括化疗和免疫疗法在内的可选疗法中，全反式维甲酸（ATRA）是维生素A的天然代谢产物，已显示出通过诱导黑色素瘤细胞分化、凋亡、生长停滞和免疫调节来治疗黑色素瘤的潜力。然而，单独使用ATRA治疗可能会导致耐药和复发。此外，鞘磷脂（SM）在黑色素瘤进展过程中参与抑制细胞增殖、分化和凋亡性细胞死亡。
基于细胞活力、凋亡性细胞死亡、细胞周期进程和基因表达水平，研究了ATRA和SM对体外B16F10黑色素瘤模型的联合抗癌作用；而在RAW264.7巨噬细胞上测试了这些治疗的安全性。
123μM的ATRA与136μM的SM联合是最有效的治疗方法，细胞增殖减少50%，在48小时内导致53.91%的凋亡性细胞死亡，并使B16F10细胞停滞于G2/M期细胞周期。虽然单独使用123μM的ATRA不会改变半胱天冬酶3和Bax基因的表达，但联合使用ATRA+SM治疗分别使基因表达水平增加了2倍和5倍。联合使用ATRA+SM治疗观察到细胞周期蛋白依赖性激酶抑制剂2A增加了13倍，同时程序性死亡配体1（PD-L1）的表达被抑制了0.5倍。
联合使用ATRA和SM疗法可能是一种有前景的黑色素瘤治疗方法，有可能提高疗效，同时降低对健康细胞的毒性。
