Коbylinska Lesya I, Klyuchivska Olga Yu, Grytsyna Iryna I, Finiuk Natalia, Panchuk Rostyslav R, Starykovych Marina O, Lehka Lilya, Lesyk Roman B, Zіmenkovsky Borys S, Stoika Rostyslav S
Rostyslav Stoika, Drahomanov Str. 14/16, 79005 Lviv, Ukraine,
Croat Med J. 2017 Apr 14;58(2):150-159. doi: 10.3325/cmj.2017.58.150.
To compare various pro-apoptotic effects of synthetic 4-thiazolidinone derivative (Les-3288), doxorubicin (Dox) and temozolomide (TMZ) in the treatment of human glioma U251 cells to improve treatment outcomes of glioblastoma and avoid anticancer drug resistance.
The cytotoxic effects of drugs used in human glioma U251 cells were measured by cell viability and proliferation assay (MTT), Trypan blue exclusion test, and Western-blot analysis of the apoptosis-related proteins. In addition, flow cytometry study of reactive oxygen species (ROS) level in glioma cells was carried out. Cytomorphological changes in treated cells were monitored by fluorescent microscopy after cell staining with Hoechst 33342 and ethydium bromide.
Half-maximal inhibitory concentration (IC50) of Les-3288, Dox, and TMZ was calculated for human glioblastoma U251 cells. The rating of the values of this indicator of cellular vitality was assessed. The results of MTT assay proved the superiority of Les-3288 vs Les-3288>Dox>TMZ, which is in agreement with the results of Trypan blue testing showing Les-3288≈Dox>TMZ. In general, such ranking corresponded to a scale of pro-apoptotic impairments in the morphology of glioma U251 cells and the results of Western-blot analysis of cleaved Caspase 3. Contrary to Dox, Les-3288 and TMZ did not affect significantly ROS levels in the treated cells.
The effect of the synthetic 4-thiazolidinone derivative Les-3288 is realized via apoptosis mechanisms and does not involve ROS. In comparison with Dox and TMZ, it is more effective in destroying human glioblastoma U251 cells. Les-3288 compound has a potential as an anticancer drug for glioblastoma. Nevertheless, further preclinical studies of the blood-brain barrier are needed.
比较合成的4-噻唑烷酮衍生物（Les-3288）、阿霉素（Dox）和替莫唑胺（TMZ）在治疗人胶质瘤U251细胞中的各种促凋亡作用，以改善胶质母细胞瘤的治疗效果并避免抗癌药物耐药性。
通过细胞活力和增殖测定（MTT）、台盼蓝排斥试验以及凋亡相关蛋白的蛋白质印迹分析，检测用于人胶质瘤U251细胞的药物的细胞毒性作用。此外，还进行了胶质瘤细胞中活性氧（ROS）水平的流式细胞术研究。在用Hoechst 33342和溴化乙锭对细胞染色后，通过荧光显微镜监测处理后细胞的细胞形态学变化。
计算了人胶质母细胞瘤U251细胞中Les-3288、Dox和TMZ的半数抑制浓度（IC50）。评估了该细胞活力指标值的等级。MTT试验结果证明Les-3288优于Les-3288>Dox>TMZ，这与台盼蓝试验结果一致，显示Les-3288≈Dox>TMZ。总体而言，这种排名与胶质瘤U251细胞形态学中的促凋亡损伤程度以及裂解的半胱天冬酶3的蛋白质印迹分析结果相对应。与Dox相反，Les-3288和TMZ对处理后细胞中的ROS水平没有显著影响。
合成的4-噻唑烷酮衍生物Les-3288的作用是通过凋亡机制实现的，不涉及ROS。与Dox和TMZ相比，它在破坏人胶质母细胞瘤U251细胞方面更有效。Les-3288化合物具有作为胶质母细胞瘤抗癌药物的潜力。然而，需要对血脑屏障进行进一步的临床前研究。
