Kakino S, Sasaki K, Kurose A, Ito H
Department of Neurosurgery, Yamaguchi University School of Medicine, Japan.
Cytometry. 1996 May 1;24(1):49-54. doi: 10.1002/(SICI)1097-0320(19960501)24:1<49::AID-CYTO6>3.0.CO;2-D.
We investigated cyclin B1 expression during the cell cycle in human glioma cells cultured under asynchronous growing condition by two cytometry techniques: flow cytometry (FCM) and laser scanning cytometry (LSC). FCM analysis revealed the specific accumulation of cyclin B1 in G2/M phase with a wide intercellular variation (Dunphy WG: Trend Cell Biol 4:202-207, 1994). It is noteworthy that LSC, which is characterized by rapid quantitative analysis followed by imaging, allows morphological observation of the intracellular distribution of cyclin B1 as a function of cell cycle position cell by cell (Hunter T: Cell 75:839-841, 1993). Cyclin B1 was virtually undetectable in cells from G0/G1 phase to mid S phase, but became visible in the cytoplasm in late S phase. As cells proceeded within G2 phase, the level of cyclin B1 rapidly increased in the perinuclear region of the cytoplasm, but cyclin B1 was still faintly present in the nucleus. Cyclin B1 appeared in the nucleus at the mitotic phase. Then the nuclear membrane was disrupted and cyclin B1 was distributed evenly in the cell. The level of cyclin B1 was maximum in metaphase. However, it abruptly degraded at the end of metaphase, and subsequently G1 cells were cyclin B1 negative.
我们采用两种细胞计数技术,即流式细胞术(FCM)和激光扫描细胞术(LSC),研究了在非同步生长条件下培养的人胶质瘤细胞在细胞周期中的细胞周期蛋白B1表达情况。FCM分析显示细胞周期蛋白B1在G2/M期有特异性积累,且细胞间差异较大(邓菲·W·G:《细胞生物学趋势》4:202 - 207,1994年)。值得注意的是,LSC以快速定量分析随后成像为特点,能够逐个细胞地观察细胞周期蛋白B1在细胞内的分布与细胞周期位置的关系(亨特·T:《细胞》75:839 - 841,1993年)。在从G0/G1期到S期中期的细胞中几乎检测不到细胞周期蛋白B1,但在S期后期其在细胞质中可见。随着细胞进入G2期,细胞周期蛋白B1在细胞质的核周区域水平迅速升高,但细胞周期蛋白B1在细胞核中仍微弱存在。在有丝分裂期细胞周期蛋白B1出现在细胞核中。然后核膜破裂,细胞周期蛋白B1在细胞内均匀分布。细胞周期蛋白B1的水平在中期最高。然而,在中期结束时它突然降解,随后G1期细胞的细胞周期蛋白B1呈阴性。