Otsuki Yoshinori, Li Zhonglian, Shibata Masa-Aki
Department of Anatomy and Biology, Osaka Medical College, 2-7 Daigaku-machi, Takatsuki, Osaka 569-8686, Japan.
Prog Histochem Cytochem. 2003;38(3):275-339. doi: 10.1016/s0079-6336(03)80002-5.
To date, many terms have been given for cell death and forgotten before they became widely accepted. Most researchers studying cell death use some terms for cell death that have survived over centuries and regard apoptosis and programmed cell death (PCD), and necrosis and oncosis as synonymous. The different terminologies used for cell death depending on the preference of researchers sometimes cause confusion in the study of apoptosis. The study of apoptosis was first based on cell morphology using transmission electron microscopy (TEM): chromatin condensation, cellular shrinkage, budding and apoptotic body formation. Recently, marked progress in biochemistry, molecular biology and genetics provided researchers of apoptosis various tools for apoptosis detection, such as the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end-labeling (TUNEL) method, agarose gel electrophoresis using extracted DNA, staining methods using fluorescence dyes, and flow cytometry. This review focuses on the following topics: history of the study of cell death and its classification, apoptosis-related proteins and their signal pathways, morphological, biochemical and molecular biological methods of apoptosis detection, and the transcriptional regulation of bcl-2 using the real-time Southwestern method and cancer gene therapy. In addition, the merits and demerits of the above-mentioned apoptosis detection methods are discussed.
迄今为止,已有许多关于细胞死亡的术语出现,但在它们被广泛接受之前就被遗忘了。大多数研究细胞死亡的人员使用了一些历经数百年沿用下来的细胞死亡术语,并将凋亡与程序性细胞死亡(PCD)以及坏死与胀亡视为同义词。因研究人员的偏好而使用的不同细胞死亡术语,有时会在凋亡研究中造成混乱。凋亡研究最初是基于透射电子显微镜(TEM)下的细胞形态学:染色质浓缩、细胞皱缩、出芽及凋亡小体形成。近来,生物化学、分子生物学及遗传学方面取得的显著进展为凋亡研究人员提供了多种凋亡检测工具,诸如末端脱氧核苷酸转移酶(TdT)介导的dUTP-生物素缺口末端标记(TUNEL)法、使用提取DNA的琼脂糖凝胶电泳、使用荧光染料的染色方法以及流式细胞术。本综述聚焦于以下主题:细胞死亡研究的历史及其分类、凋亡相关蛋白及其信号通路、凋亡检测的形态学、生物化学及分子生物学方法,以及运用实时西南杂交法对bcl-2的转录调控和癌症基因治疗。此外,还讨论了上述凋亡检测方法的优缺点。