Proapoptotic effects of novel thiazole derivative on human glioma cells

Abstract

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.

Effects of thiazole derivatives on intracellular structure and functions in murine lymphoma cells

“…In our previous studies, we found that the novel thiazole derivatives BF1 and PP2 activate caspasedependent and mitochondria-associated mechanism of apoptosis [6,7]. In order to reveal potential morphological changes in mitochondria in NK/Ly lymphoma cells under treatment with BF1 and PP2, we have carried out the electron microscopy study of these cells.…”

“…These compounds also had low toxicity towards human kidney cells (HEK 293) and human keratinocytes (HaCaT line) [5,8]. Previously we have identified that BF1 increased the level of Bax and Bim pro-apoptotic proteins in glioma cells [6], and PP2 increased the level of pro-apoptotic Bim protein and the mitochondriaspecific EndoG nuclease, and decreased the level of the anti-apoptotic Bcl-2 protein in the leukemia cells in vitro [7].…”

“…introduction Thiazole derivatives are attractive heterocycles for pharmaceutical and medicinal chemists who design new potent anticancer agents [1][2][3][4]. It was shown that two novel thiazole derivatives -N-(5-benzyl-1,3-thiazol-2-yl)-3,5-dimethyl-1-benzofuran-2-carboxamide (BF1) and 7-benzyl-8-methyl-2-propylpyrazolo[4,3-e]thiazolo [3,2-a] pyrimidin-4(2H)-one (PP2) [5] have a high cytotoxic effect toward various cancer cell lines, such as glioblastoma (U251 and T98G) and human myeloid leukemia (HL 60 and K562) [6,7]. These compounds also had low toxicity towards human kidney cells (HEK 293) and human keratinocytes (HaCaT line) [5,8].…”

Apoptosis induction in human leukemia cells by novel 2-amino-5-benzylthiazole derivatives

“…For calculation of the olive tail mo-ment (OTL), Casplab-1.2.3b2 software (CASPLab, Wroclaw, Poland) was used. OTL was studied as a product of the detectable DNA in the comet tail, and distance between centers of the head mass and tail region, and calculated as OTL = (Tail mean -Head mean)×% of tail DNA [9,10].…”

“…Gel retardation assay of plasmid DNA. Plasmid DNA (1 μg/μl) pEGFPc-1 (Clontech, Mountain View, California, USA) was mixed with 2 μl of compounds 1, 2 (0.09, 1.0, 10.0 μM), and Dox (0.09 μM) in 18 µl of 0.9% sodium chloride (Arterium, Lviv, Ukraine) at room temperature for 1 h. The prepared mixture was analyzed by electrophoresis in 1% agarose gel (Lachema, Brno, Czech Republic) with 1× Tris acetate (TAE) buffer containing 1 μg/ml of EtBr (Sigma-Aldrich, St. Louis, Missouri, USA) for 1 h at a constant voltage of 70 V. A trans-illuminator (MacroVue UV-20, Hoeffer Pharmacia Biotech Inc., San Francisco, California, USA) was used for visualization of plasmid DNA [10].…”