Department of Diagnostic Pathology, Kobe University Hospital, Japan.
Am J Surg Pathol. 2012 May;36(5):769-73. doi: 10.1097/PAS.0b013e3182478b93.
Carcinogenesis is widely believed to occur when regulatory systems governing cellular proliferation and differentiation are compromised. To date, various methods have been devised to determine cell cycle. However, these methods have not gained popularity in the diagnostic field. We developed a multiplex immunohistochemical method that can simultaneously stain cells in the G1 and S/G2/M phases and those undergoing apoptosis with the 3 markers Cdt1, geminin, and gamma H2A.X. The staining procedure can be performed using an autoimmunostainer. The nuclei of cells in the G1 phase stain red with the antibody for Cdt1, those in the S/G2/M phases stain blue with the antibody for geminin, and the nuclei of cells undergoing apoptosis stain brown with the antibody for H2A.X. The present method enables accurate cell cycle assessments using paraffin-embedded tissue specimens, which are superior to other forms of specimens in terms of morphologic observation.
癌症发生被广泛认为是在调控细胞增殖和分化的系统受到损害时发生的。迄今为止，已经设计出了各种方法来确定细胞周期。然而，这些方法在诊断领域并没有得到普及。我们开发了一种多重免疫组织化学方法，可以同时用 Cdt1、geminin 和 γH2AX 这 3 种标志物对 G1 期和 S/G2/M 期的细胞以及正在凋亡的细胞进行染色。染色过程可以使用自动免疫染色仪进行。用 Cdt1 抗体对 G1 期的细胞核进行红色染色，用 geminin 抗体对 S/G2/M 期的细胞核进行蓝色染色，用 H2AX 抗体对正在凋亡的细胞核进行棕色染色。本方法能够使用石蜡包埋组织标本进行准确的细胞周期评估，在形态学观察方面优于其他形式的标本。
