Tabandeh Fatemeh, Moradian Tehrani Rana, Sharifi Mohammadreza, Toopchi Elmira
Medical Biotechnology. Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Department of Genetics and Molecular Biology. School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Iran J Basic Med Sci. 2025;28(6):772-783. doi: 10.22038/ijbms.2025.84187.18208.
Melanoma is one of the most aggressive and deadly skin cancers. Despite advances, effective melanoma treatment is challenging, often requiring a shift from individual therapies to combination approaches. This study explores whether combining dacarbazine (DTIC) and temozolomide (TMZ) with the siRNA approach holds promise for melanoma treatment.
To determine the IC values of DTIC and TMZ, the A375 cell line was treated with different drug concentrations for 24-72 hr. The best exposure time of BRAF siRNA transfection was performed. Subsequently, cell viability (using the MTT assay), apoptosis (by flow cytometry), and gene expression levels of B-Raf proto-oncogene, serine/threonine kinase (), caspase 3 (), and phosphoinositide-3-kinase regulatory subunit 3 () genes (by quantitative real-time PCR) were assessed in the treated groups (i.e., control, negative controls, DTIC alone, TMZalone, DTIC+ TMZ, BRAF(V600E)siRNA alone, siRNA+ DTIC, siRNA+ TMZ, and siRNA+ DTIC+ TMZ groups).
Cell viability significantly decreased in the chemotherapy-only and siRNA+drug groups, although no difference was observed between them. The apoptosis percentage in all treated groups indicated a significant difference compared to the control group. The expression of the gene notably decreased in the BRAF () siRNA +drug groups compared to the chemotherapy groups. Despite overexpression of in the chemotherapy-treated groups, the most effective enhancement was noted in the siRNA+DTIC+TMZ group (<0.0001). The mean expression of the gene in siRNA+chemotherapy groups revealed a notable reduction.
These findings suggest that the siRNA-transfected treatment groups have the potential to provide therapeutic effects comparable to those of chemotherapy.
黑色素瘤是最具侵袭性和致命性的皮肤癌之一。尽管取得了进展，但有效的黑色素瘤治疗仍具有挑战性，通常需要从单一疗法转向联合治疗方法。本研究探讨将达卡巴嗪（DTIC）和替莫唑胺（TMZ）与小干扰RNA（siRNA）方法联合应用于黑色素瘤治疗是否具有前景。
为确定DTIC和TMZ的半数抑制浓度（IC值），用不同药物浓度处理A375细胞系24至72小时。进行BRAF siRNA转染的最佳暴露时间实验。随后，在各治疗组（即对照组、阴性对照组、单独使用DTIC组、单独使用TMZ组、DTIC + TMZ组、单独使用BRAF（V600E）siRNA组、siRNA + DTIC组、siRNA + TMZ组以及siRNA + DTIC + TMZ组）中评估细胞活力（采用MTT法）、细胞凋亡（通过流式细胞术）以及B-Raf原癌基因、丝氨酸/苏氨酸激酶（BRAF）、半胱天冬酶3（caspase 3）和磷脂酰肌醇-3-激酶调节亚基3（PIK3R3）基因的表达水平（通过定量实时聚合酶链反应）。
仅化疗组和siRNA + 药物组的细胞活力均显著降低，尽管两组之间未观察到差异。所有治疗组的细胞凋亡百分比与对照组相比均有显著差异。与化疗组相比，BRAF（V600E）siRNA + 药物组中BRAF基因的表达显著降低。尽管化疗治疗组中PIK3R3基因过表达，但在siRNA + DTIC + TMZ组中观察到最有效的增强（P < 0.0001）。siRNA + 化疗组中PIK3R3基因的平均表达显著降低。
这些发现表明，siRNA转染治疗组有可能提供与化疗相当的治疗效果。
