Department of Histology, Faculty of Medicine, Medical University of Gdansk, 80‑211 Gdansk, Poland.
School of Molecular Sciences, Faculty of Science, The University of Western Australia, Perth, Western Australia 6009, Australia.
Int J Oncol. 2019 Apr;54(4):1481-1495. doi: 10.3892/ijo.2019.4725. Epub 2019 Feb 25.
Melanoma represents a significant challenge in cancer treatment due to the high drug resistance of melanomas and the patient mortality rate. This study presents data indicating that nanomolar concentrations of the hormonally active form of vitamin D, 1α,25‑dihydroxyvitamin D3 [1α,25(OH)2D3], its non‑calcemic analogues 20S‑hydroxyvitamin D3 and 21‑hydroxypregnacalciferol, as well as the low‑calcemic synthetic analog calcipotriol, modulate the efficacy of the anticancer drugs cisplatin and dacarbazine. It was observed that vitamin D analogs sensitized melanoma A375 cells to hydrogen peroxide used as an inducer of oxidative stress. On the other hand, only 1α,25(OH)2D3 resulted in a minor, but significant effect on the proliferation of melanoma cells treated simultaneously with dacarbazine, but not cisplatin. Notably, cisplatin (300 µM) exhibited a higher overall antiproliferative activity than dacarbazine. Cisplatin treatment of melanoma cells resulted in an induction of apoptosis as demonstrated by flow cytometry (accumulation of cells at the subG1 phase of the cell cycle), whereas dacarbazine caused G1/G0 cell cycle arrest, with the effects being improved by pre‑treatment with vitamin D analogs. Treatment with cisplatin resulted in an initial increase in the level of reactive oxygen species (ROS). Dacarbazine caused transient stimulation of ROS levels and the mitochondrial membrane potential (Δψm) (after 1 or 3 h of treatment, respectively), but the effect was not detectable following prolonged (24 h) incubation with the drug. Vitamin D exhibited modulatory effects on the cells treated with dacarbazine, decreasing the half maximal inhibitory concentration (IC50) for the drug, stimulating G1/G0 arrest and causing a marked decrease in Δψm. Finally, cisplatin, dacarbazine and 1α,25(OH)2D3 displayed modulatory effects on the expression of ROS and vitamin D‑associated genes in the melanoma A375 cells. In conclusion, nanomolar concentrations of 1,25(OH)2D3 only had minor effects on the proliferation of melanoma cells treated with dacarbazine, decreasing the relative IC50 value. However, co‑treatment with vitamin D analogs resulted in the modulation of cell cycle and ROS responses, and affected gene expression, suggesting possible crosstalk between the signaling pathways of vitamin D and the anticancer drugs used in this study.
黑色素瘤是癌症治疗中的一个重大挑战,这是由于黑色素瘤的药物耐药性和患者死亡率高。本研究提供的数据表明,激素活性形式的维生素 D,1α,25-二羟维生素 D3[1α,25(OH)2D3]、其非钙调素类似物 20S-羟维生素 D3 和 21-羟孕甾烷二氢胆钙化醇,以及低钙合成类似物卡泊三醇,调节了顺铂和达卡巴嗪等抗癌药物的疗效。研究发现,维生素 D 类似物使黑色素瘤 A375 细胞对用作诱导氧化应激的过氧化氢敏感。另一方面,只有 1α,25(OH)2D3 对同时用达卡巴嗪处理的黑色素瘤细胞的增殖产生轻微但显著的影响,但对顺铂没有影响。值得注意的是,顺铂(300μM)对黑色素瘤细胞的总体增殖抑制活性高于达卡巴嗪。顺铂处理黑色素瘤细胞导致凋亡的诱导,如通过流式细胞术(细胞周期的 subG1 期细胞的积累)所示,而达卡巴嗪引起 G1/G0 细胞周期阻滞,用维生素 D 类似物预处理可改善这种效果。用顺铂处理导致活性氧(ROS)水平的初始增加。达卡巴嗪引起 ROS 水平和线粒体膜电位(Δψm)的短暂刺激(分别在处理 1 或 3 小时后),但在用药物孵育 24 小时后,这种作用无法检测到。维生素 D 对用达卡巴嗪处理的细胞具有调节作用,降低了药物的半最大抑制浓度(IC50),刺激 G1/G0 阻滞,并导致 Δψm 显著下降。最后,顺铂、达卡巴嗪和 1α,25(OH)2D3 对黑色素瘤 A375 细胞中 ROS 和维生素 D 相关基因的表达均有调节作用。总之,纳米摩尔浓度的 1,25(OH)2D3 对用达卡巴嗪处理的黑色素瘤细胞的增殖仅有轻微影响,降低了相对 IC50 值。然而,与维生素 D 类似物共同处理导致细胞周期和 ROS 反应的调节,并影响基因表达,这表明维生素 D 的信号通路与本研究中使用的抗癌药物之间可能存在串扰。