Aktug Huseyin, Acikgoz Eda, Uysal Aysegul, Oltulu Fatih, Oktem Gulperi, Yigitturk Gurkan, Demir Kenan, Yavasoglu Altug, Bozok Cetintas Vildan
Department of Histology and Embryology, Faculty of Medicine, Ege University, Izmir, Turkey.
Department of Histology and Embryology, Faculty of Medicine, Yuzuncu Yil University, Van, Turkey.
Tumour Biol. 2016 Sep;37(9):12423-12440. doi: 10.1007/s13277-016-5108-9. Epub 2016 Jun 21.
Similarities and differences in the cell cycle components, apoptosis and cytoskeleton-related molecules among mouse skin fibroblast cells (MSFs), mouse squamous cell lung carcinomas (SqCLCs) and mouse embryonic stem cells (mESCs) are important determinants of the behaviour and differentiation capacity of these cells. To reveal apoptotic pathways and to examine the distribution and the role of cell cycle-cell skeleton comparatively would necessitate tumour biology and stem cell biology to be assessed together in terms of oncogenesis and embryogenesis. The primary objectives of this study are to investigate the effects of flavopiridol, a cell cycle inhibitor, and geldanamycin, a heat shock protein inhibitor on mouse somatic, tumour and embryonic stem cells, by specifically focusing on alterations in cytoskeletal proteins, cell polarity and motility as well as cell cycle regulators. To meet these objectives, expression of several genes, cell cycle analysis and immunofluorescence staining of intracellular cytoskeletal molecules were performed in untreated and flavopiridol- or geldanamycin-treated cell lines. Cytotoxicity assays showed that SqCLCs are more sensitive to flavopiridol than MSFs and mESCs. Keratin-9 and keratin-2 expressions increased dramatically whereas cell cycle regulatory genes decreased significantly in the flavopiridol-treated MSFs. Flavopiridol-treated SqCLCs displayed a slight increase in several cell cytoskeleton regulatory genes as well as cell cycle regulatory genes. However, gene expression profiles of mESCs were not affected after flavopiridol treatment except the Cdc2a. Cytotoxic concentrations of geldanamycin were close to each other for all cell lines. Cdkn1a was the most increased gene in the geldanamycin-treated MSFs. However, expression levels of cell cytoskeleton-associated genes were increased dramatically in the geldanamycin-treated SqCLCs. Our results revealing differences in molecular mechanisms between embryogenesis and carcinogenesis may prove crucial in developing novel therapeutics that specifically target cancer cells.
小鼠皮肤成纤维细胞（MSFs）、小鼠肺鳞状细胞癌（SqCLCs）和小鼠胚胎干细胞（mESCs）在细胞周期成分、细胞凋亡和细胞骨架相关分子方面的异同是这些细胞行为和分化能力的重要决定因素。为了揭示细胞凋亡途径，并比较细胞周期与细胞骨架的分布及作用，有必要从肿瘤发生和胚胎发生的角度共同评估肿瘤生物学和干细胞生物学。本研究的主要目的是研究细胞周期抑制剂黄酮哌啶醇和热休克蛋白抑制剂格尔德霉素对小鼠体细胞、肿瘤细胞和胚胎干细胞的影响，特别关注细胞骨架蛋白、细胞极性和运动性以及细胞周期调节因子的变化。为实现这些目标，在未处理以及经黄酮哌啶醇或格尔德霉素处理的细胞系中进行了几种基因的表达、细胞周期分析和细胞内细胞骨架分子的免疫荧光染色。细胞毒性试验表明，SqCLCs对黄酮哌啶醇比MSFs和mESCs更敏感。在经黄酮哌啶醇处理的MSFs中，角蛋白9和角蛋白2的表达显著增加，而细胞周期调节基因显著减少。经黄酮哌啶醇处理的SqCLCs在几种细胞骨架调节基因以及细胞周期调节基因方面略有增加。然而，除Cdc2a外，黄酮哌啶醇处理后mESCs的基因表达谱未受影响。所有细胞系中格尔德霉素的细胞毒性浓度彼此接近。Cdkn1a是经格尔德霉素处理的MSFs中增加最多的基因。然而，在经格尔德霉素处理的SqCLCs中，细胞骨架相关基因的表达水平显著增加。我们揭示胚胎发生和肿瘤发生之间分子机制差异的结果可能对开发特异性靶向癌细胞的新型疗法至关重要。
