Institute of Cell Biology of National Academy of Sciences of Ukraine.
Biology Faculty.
Anticancer Drugs. 2019 Jan;30(1):27-37. doi: 10.1097/CAD.0000000000000686.
The aim of the present study was to investigate the antiproliferative and proapoptotic actions of N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide derivative (compound 5) in glioma cells in comparison with the actions of temozolomide (TMZ) and doxorubicin (Dox), used as positive controls. The antiproliferative activity of the compound 5, TMZ, and Dox on human glioblastoma U251 and human glioblastoma multiform T98G cells was measured using the MTT test. Western blot analysis, fluorescent microscopy, agarose gel retardation assay, flow cytometric analysis, and the DNA comet assay under alkaline conditions were carried out to study the effect of compound 5 on U251 cells. This compound showed ~20 times higher cytotoxicity toward U251 and T98G cells compared with the effects of TMZ and approximately two times higher activity than that of the Dox. Compound 5 induced apoptosis in U251 cells by PARP1 and caspase 3 cleavage mechanisms, also inducing an increase in the level of Bax and Bim proapoptotic proteins and a decrease in the level of phosho-ERK1/2 kinase. The cytotoxicity of compound 5 was associated with an increase in the production of the hydrogen peroxide and the formation of DNA single-strand breaks. This compound 5 did not intercalate into a DNA molecule. Thus, the novel thiazole derivative (compound 5) proved to be a potential antiglioma drug that showed much higher cytotoxic action on human glioma cells compared with the effects of TMZ and Dox. Its cytotoxicity is associated with apoptosis induction, production of the reactive oxygen species, and formation of DNA single-strand breaks without significant DNA intercalation.
本研究旨在研究 N-(5-苄基-1,3-噻唑-2-基)-3,5-二甲基-1-苯并呋喃-2-甲酰胺衍生物（化合物 5）在胶质瘤细胞中的抗增殖和促凋亡作用，并与替莫唑胺（TMZ）和多柔比星（Dox）的作用进行比较，后者被用作阳性对照。使用 MTT 试验测量化合物 5、TMZ 和 Dox 对人神经胶质瘤 U251 和人多形性胶质母细胞瘤 T98G 细胞的增殖活性。通过 Western blot 分析、荧光显微镜、琼脂糖凝胶阻滞试验、流式细胞术分析和碱性条件下的 DNA 彗星试验研究化合物 5 对 U251 细胞的影响。与 TMZ 和 Dox 的作用相比，该化合物对 U251 和 T98G 细胞的细胞毒性高约 20 倍，对 U251 细胞的活性高约 2 倍。化合物 5 通过 PARP1 和 caspase 3 切割机制诱导 U251 细胞凋亡，同时诱导促凋亡蛋白 Bax 和 Bim 的水平增加，磷酸化 ERK1/2 激酶的水平降低。化合物 5 的细胞毒性与过氧化氢的产生和 DNA 单链断裂的形成有关。该化合物 5 不嵌入 DNA 分子。因此，新型噻唑衍生物（化合物 5）被证明是一种潜在的抗神经胶质瘤药物，与 TMZ 和 Dox 的作用相比，对人神经胶质瘤细胞具有更高的细胞毒性作用。其细胞毒性与凋亡诱导、活性氧的产生和 DNA 单链断裂的形成有关，而没有明显的 DNA 嵌入。
