Skobowiat Cezary, Oak Allen S W, Kim Tae-Kang, Yang Chuan He, Pfeffer Lawrence M, Tuckey Robert C, Slominski Andrzej T
Department of Dermatology, University of Alabama at Birmingham, AL, USA.
Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Collegium Medicum, Nicolaus Copernicus University in Torun, Poland.
Oncotarget. 2017 Feb 7;8(6):9823-9834. doi: 10.18632/oncotarget.14193.
A novel pathway of vitamin D3 (D3) metabolism, initiated by C20-hydroxylation of D3 by CYP11A1, has been confirmed to operate in vivo. Its major product, 20(OH)D3, exhibits antiproliferative activity in vitro comparable to that of 1,25(OH)2D3, but is noncalcemic in mice and rats. To further characterize the antimelanoma activity of 20(OH)D3, we tested its effect on colony formation of human melanoma cells in monolayer culture and anchorage-independent growth in soft agar. The migratory capabilities of the cells and cell-cell and cell-extracellular matrix interactions were also evaluated using transwell cell migration and spheroid toxicity assays. To assess the antimelanoma activity of 20(OH)D3in vivo, age-matched immunocompromised mice were subcutaneously implanted with luciferase-labelled SKMel-188 cells and were randomly assigned to be treated with either 20(OH)D3 or vehicle (n=10 per group). Tumor size was measured with caliper and live bioimaging methods, and overall health condition expressed as a total body score scale. The following results were observed: (i) 20(OH)D3 inhibited colony formation both in monolayer and soft agar conditions, (ii) 20(OH)D3 inhibited melanoma cells in both transwell migration and spheroid toxicity assays, and (iii) 20(OH)D3 inhibited melanoma tumor growth in immunocompromised mice without visible signs of toxicity. However, although the survival rate was 90% in both groups, the total body score was higher in the treatment group compared to control group (2.8 vs. 2.55). In conclusion, 20(OH)D3, an endogenously produced secosteroid, is an excellent candidate for further preclinical testing as an antimelanoma agent.
一种由CYP11A1将维生素D3（D3）进行C20-羟基化引发的新型D3代谢途径已在体内得到证实。其主要产物20（OH）D3在体外表现出与1,25（OH）2D3相当的抗增殖活性，但在小鼠和大鼠中无血钙升高作用。为了进一步表征20（OH）D3的抗黑色素瘤活性，我们测试了其对单层培养的人黑色素瘤细胞集落形成以及软琼脂中不依赖贴壁生长的影响。还使用Transwell细胞迁移和球体毒性试验评估了细胞的迁移能力以及细胞-细胞和细胞-细胞外基质相互作用。为了评估20（OH）D3在体内抗黑色素瘤的活性，将年龄匹配的免疫受损小鼠皮下植入荧光素酶标记的SKMel-188细胞，并随机分为两组，分别用20（OH）D3或赋形剂处理（每组n = 10）。用卡尺和活体生物成像方法测量肿瘤大小，并用全身评分量表表示整体健康状况。观察到以下结果：（i）20（OH）D3在单层和软琼脂条件下均抑制集落形成，（ii）20（OH）D3在Transwell迁移和球体毒性试验中均抑制黑色素瘤细胞，（iii）20（OH）D3在免疫受损小鼠中抑制黑色素瘤肿瘤生长，且无明显毒性迹象。然而，尽管两组的存活率均为90%，但治疗组的全身评分高于对照组（2.8对2.55）。总之，内源性产生的甾醇类化合物20（OH）D3作为一种抗黑色素瘤药物，是进一步临床前试验的极佳候选物。
