Jo Deuk-Won, Kim Young-Kyun, Yun Pil-Young
Department of Prosthodontics, Section of Dentistry, Seoul National University Bundang Hospital, Seongnam, Korea.
Department of Oral and Maxillofacial Surgery, Section of Dentistry, Seoul National University Bundang Hospital, Seongnam, Korea.
J Korean Assoc Oral Maxillofac Surg. 2016 Dec;42(6):337-344. doi: 10.5125/jkaoms.2016.42.6.337. Epub 2016 Dec 27.
The purpose of this study was to evaluate the anti-cancer activity of cisplatin by studying its effects on cell viability and identifying the mechanisms underlying the induction of cell cycle arrest and apoptosis on oral squamous cell carcinoma (OSCC) cell lines with varying p53 mutation status.
Three OSCC cell lines, YD-8 (p53 point mutation), YD-9 (p53 wild type), and YD-38 (p53 deletion) were used. To determine the cytotoxic effect of cisplatin, MTS assay was performed. The cell cycle alteration and apoptosis were analyzed using flow cytometry. Western blot analysis was used to detect the expression of cell cycle alteration- or apoptosis-related proteins as well as p53.
Cisplatin showed a time- and dose-dependent anti-proliferative effect in all cell lines. Cisplatin induced G2/M cell accumulation in the three cell lines after treatment with 0.5 and 1.0 µg/mL of cisplatin for 48 hours. The proportion of annexin V-FITC-stained cells increased following treatment with cisplatin. The apoptotic proportion was lower in the YD-38 cell line than in the YD-9 or YD-8 cell lines. Also, immunoblotting analysis indicated that p53 and p21 were detected only in YD-8 and YD-9 cell lines after cisplatin treatment.
In this study, cisplatin showed anti-cancer effects via G2/M phase arrest and apoptosis, with some difference among OSCC cell lines. The mutation status of p53 might have influenced the difference observed among cell lines. Further studies on p53 mutation status are needed to understand the biological behavior and characteristics of OSCCs and to establish appropriate treatment.
本研究旨在通过研究顺铂对细胞活力的影响，并确定其在不同p53突变状态的口腔鳞状细胞癌（OSCC）细胞系中诱导细胞周期停滞和凋亡的潜在机制，来评估顺铂的抗癌活性。
使用三种OSCC细胞系，即YD-8（p53点突变）、YD-9（p53野生型）和YD-38（p53缺失）。为了确定顺铂的细胞毒性作用，进行了MTS测定。使用流式细胞术分析细胞周期改变和凋亡情况。蛋白质免疫印迹分析用于检测细胞周期改变或凋亡相关蛋白以及p53的表达。
顺铂在所有细胞系中均表现出时间和剂量依赖性的抗增殖作用。在用0.5和1.0μg/mL顺铂处理48小时后，顺铂在三种细胞系中诱导了G2/M期细胞积累。顺铂处理后，膜联蛋白V-FITC染色细胞的比例增加。YD-38细胞系中的凋亡比例低于YD-9或YD-8细胞系。此外，免疫印迹分析表明，顺铂处理后仅在YD-8和YD-9细胞系中检测到p53和p21。
在本研究中，顺铂通过G2/M期阻滞和凋亡显示出抗癌作用，在OSCC细胞系之间存在一些差异。p53的突变状态可能影响了细胞系之间观察到的差异。需要进一步研究p53突变状态，以了解OSCC的生物学行为和特征，并建立适当的治疗方法。
