Mainwaring P N, Ellis P A, Detre S, Smith I E, Dowsett M
Academic Department of Biochemistry, Royal Marsden Hospital, London, UK.
J Clin Pathol. 1998 Jan;51(1):34-7. doi: 10.1136/jcp.51.1.34.
Apoptosis has a role in many cellular processes including development, normal tissue homeostasis, and malignancy. This aspect of research is relatively new with distinct methods of analysing disparate biochemical and genetic events to measure apoptotic cells. The use of biotinylated nucleotides to identify DNA strand breaks is a commonly reported method of estimating cells numbers undergoing apoptosis; however, investigators report inconsistent results for a variety of reasons.
To compare two in situ techniques of measuring apoptosis: in situ nick translation (ISNT) and TdT mediated dUTP-biotin nick end labelling (TUNEL); and to assess DNA cleavage in 20 paired paraffin wax embedded breast cancer tissues from patients; one group who had received no prior treatment and one group who had received chemohormonal treatment.
Apoptotic scores obtained from paraffin wax embedded human breast cancer after using ISNT and TUNEL methods were not significantly different (p = 0.11). A strong correlation between scores obtained from the two techniques was found (r = 0.758, p < 0.0001). Optimisation of both techniques is crucial to ensure maximal assay performance in breast cancer tissue.
细胞凋亡在包括发育、正常组织稳态和恶性肿瘤在内的许多细胞过程中发挥作用。这方面的研究相对较新，有不同的方法来分析不同的生化和基因事件以检测凋亡细胞。使用生物素化核苷酸来识别DNA链断裂是一种常用的估计凋亡细胞数量的方法；然而，由于各种原因，研究人员报告的结果并不一致。
比较两种测量细胞凋亡的原位技术：原位缺口平移（ISNT）和末端脱氧核苷酸转移酶介导的dUTP生物素缺口末端标记（TUNEL）；并评估20对来自患者的石蜡包埋乳腺癌组织中的DNA切割情况；一组未接受过任何治疗，另一组接受了化学激素治疗。
使用ISNT和TUNEL方法从石蜡包埋的人乳腺癌中获得的凋亡评分没有显著差异（p = 0.11）。发现两种技术获得的评分之间有很强的相关性（r = 0.758，p < 0.0001）。两种技术的优化对于确保在乳腺癌组织中获得最大的检测性能至关重要。
