Mohamed Hadeel A B Elnaim, Agus Hizlan Hincal, Palabiyik Bedia
Institute of Graduate Studies in Science, Department of Molecular Biology and Genetics, Istanbul University, 34116, Istanbul, Turkey.
Faculty of Science and Literature, Department of Molecular Biology and Genetics, Istanbul Yeni Yuzyil University, 34010, Istanbul, Turkey.
FEMS Yeast Res. 2025 Jan 30;25. doi: 10.1093/femsyr/foae040.
Fission yeast is the ideal model organism for studying telomere maintenance in higher eukaryotes. Telomere length has been directly correlated with life expectancy and the onset of aging-related diseases in mammals. In this study, we developed a novel simple, and reproducible method to measure the telomere length, by investigating the effect of caffeine and cisplatin on the telomere length in fission yeast. Hydroxyurea-synchronized fission yeast cells were exposed to 62 µM cisplatin and 8.67 mM caffeine treatments for 2 h, then their telomere lengths were evaluated with two different methods. First, the quantitative polymerase chain reaction (qPCR) assay was used as a confirmative method, where telomere length was determined relative to a single-copy gene in the genome. Second, the newly developed method standard polymerase chain reaction (PCR)/ImageJ assay assessed the telomere length based on the amplified PCR band intensity using a set of telomere primers, reflecting telomeric sequence availability in the genome. Both methods show a significant decrease and a notable telomere lengthening in response to cisplatin and caffeine treatments, respectively. The finding supports the accuracy and productivity of the standard PCR/ImageJ assay as it can serve as a quick screening tool to study the effect of suspected chemotherapeutic and antiaging drugs on telomere length in fission yeast.
裂殖酵母是研究高等真核生物端粒维持的理想模式生物。端粒长度与哺乳动物的预期寿命和衰老相关疾病的发生直接相关。在本研究中，我们通过研究咖啡因和顺铂对裂殖酵母端粒长度的影响，开发了一种新颖、简单且可重复的端粒长度测量方法。用羟基脲同步化的裂殖酵母细胞分别用62µM顺铂和8.67mM咖啡因处理2小时，然后用两种不同的方法评估其端粒长度。首先，定量聚合酶链反应（qPCR）测定法用作确认方法，其中端粒长度相对于基因组中的单拷贝基因进行测定。其次，新开发的标准聚合酶链反应（PCR）/ImageJ测定法使用一组端粒引物，根据扩增的PCR条带强度评估端粒长度，反映基因组中端粒序列的可用性。两种方法均显示，顺铂和咖啡因处理分别导致端粒显著缩短和显著延长。这一发现支持了标准PCR/ImageJ测定法的准确性和高效性，因为它可作为一种快速筛选工具，用于研究疑似化疗药物和抗衰老药物对裂殖酵母端粒长度的影响。
