Effects of clotrimazole on the growth, morphological characteristics, and cisplatin sensitivity of human glioblastoma cells in vitro
Effects of clotrimazole on the growth, morphological characteristics, and cisplatin sensitivity of human glioblastoma cells in vitro
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DOI：
10.3171/jns.1999.90.5.0918
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发表时间：
1999-05-01
影响因子：
4.1
通讯作者：
Hiura, T
中科院分区：
文献类型：
--
作者：
Khalid, MH;Shibata, S;Hiura, T
Object.克霉唑是一种抗真菌药物，通过下调细胞内Ca++和K+的运动来抑制正常细胞和癌细胞的增殖。作者在体外研究了克霉唑对两种人胶质母细胞瘤细胞系A172（具有野生型p53基因）和T98 G（具有突变型p53基因）的生长和对顺铂的敏感性的影响。将A172和T98 G胶质母细胞瘤细胞暴露于克霉唑，并使用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基氯化四唑比色测定法评估细胞生长。克霉唑对细胞增殖产生剂量依赖性抑制，并导致细胞结构向分化良好的形式变化。克霉唑的生长抑制作用是可逆的。Western免疫印迹分析显示细胞胶质细胞酸性蛋白和野生型p53显着增加，c-myc和c-fos癌蛋白在克霉唑治疗的两个细胞系的减少。流式细胞术分析显示，克霉唑处理的细胞在G（0）/G（i）期积累，S期细胞显著减少;当克霉唑从培养基中洗出时，细胞再次开始增殖，G（0）/G（i）期细胞显著减少，S期细胞增加。克霉唑的生长抑制作用不能被表皮生长因子或c-myc肽的外源性刺激所克服。与单独使用任一种药物的治疗相比，克霉唑和顺铂的联合治疗显著增强了细胞毒性。DNA片段化实验表明，克霉唑和顺铂均诱导细胞凋亡，且两种药物处理的细胞凋亡率均增加。结论。目前的研究表明，克霉唑抑制细胞增殖伴随着形态学的变化，向胶质母细胞瘤细胞的分化，这种药物协同增强顺铂的抗肿瘤作用，诱导野生型p53介导的凋亡。
Object. Clotrimazole, an antimycotic drug, inhibits proliferation of normal and cancer cells by downregulating the movement of intracellular Ca++ and K+. The authors examined the effect of clotrimazole on the growth and sensitivity to cisplatin of two human glioblastoma cell lines-A172, which has the wild-type p53 gene, and T98G, which has the mutant p53 gene in vitro.Methods. The A172 and T98G glioblastoma cells were exposed to clotrimazole and cell growth was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium chloride colorimetric assay. Clotrimazole produced a dose-dependent inhibition of cell proliferation and caused changes in cellular structure toward a well-differentiated form. The growth inhibitory effect of clotrimazole was reversible. Western immunoblot analysis revealed a marked increase in cellular glial fibrillary acidic protein and wild-type p53 and a decrease in c-myc and c-fos oncoproteins in both cell lines treated with clotrimazole. Flow cytometric analysis revealed that clotrimazole-treated cells accumulated in the G(0)/G(i) phase with a marked decrease in cells in the S phase; when clotrimazole was washed out from the culture medium, cells again started to proliferate, with a marked decrease in cells in the G(0)/G(i) phase and an increase in cells in the S phase. The growth inhibitory effect of clotrimazole could not be overcome by exogenous stimulation with either epidermal growth factor or c-myc peptide. A combined treatment with clotrimazole and cisplatin significantly enhanced cell cytotoxicity compared with treatment using either drug alone. A DNA fragmentation assay showed that both clotrimazole and cisplatin induced apoptosis, which was increased in cells treated by both drugs.Conclusions. The present study indicates that clotrimazole inhibits cell proliferation accompanied by morphological changes toward differentiation of glioblastoma cells and that this drug synergistically enhances the antitumor effect of cisplatin by inducing wild-type p53-mediated apoptosis.