Suetsugu Atsushi, Jiang Ping, Yang Meng, Yamamoto Norio, Moriwaki Hisataka, Saji Shigetoyo, Hoffman Robert M
AntiCancer Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. Department of Gastroenterology, Gifu University Graduate School of Medicine, Gifu, Japan.
AntiCancer Inc., San Diego, CA, U.S.A.
Anticancer Res. 2015 May;35(5):2553-7.
BACKGROUND/AIM: A library of dual-color fluorescent cancer cells with green fluorescent protein (GFP), linked to histone H2B, expressed in the nucleus and red fluorescent protein (RFP) expressed in the cytoplasm was previously genetically engineered. The aim of the current study was to use the dual-color cancer cells to visualize chromosome and cytoplasmic dynamics during mitosis.
Using an Olympus FV1000 confocal microscope, a library of dual-color cells from the major cancer types was cultured on plastic. The cells were imaged by confocal microscopy to demonstrate chromosome and cytoplasmic dynamics during mitosis.
Nuclear GFP expression enabled visualization of chromosomes behavior, whereas simultaneous cytoplasmic RFP expression enabled visualization of cytoplasmic behavior during mitosis. Thus, total cellular dynamics can be visualized at high resolution, including individual chromosomes in some cases, in living dual-color cells in real time.
Dual-color cancer cells expressing H2B-GFP in the nucleus and RFP in the cytoplasm provide unique tools for visualizing subcellular nuclear and cytoplasm dynamics, including the behavior of individual chromosomes during mitosis. The dual-color cells can be used to evaluate chromosomal loss or gain in real time during treatment with a variety of agents or as the cells are selected for increased or decreased malignancy in culture or in vivo. The dual color cells will be a useful tool to discover and evaluate novel strategies for killing cancer cells.
背景/目的：先前已通过基因工程构建了一个双色荧光癌细胞库，其中与组蛋白H2B相连的绿色荧光蛋白（GFP）在细胞核中表达，红色荧光蛋白（RFP）在细胞质中表达。本研究的目的是利用双色癌细胞观察有丝分裂过程中的染色体和细胞质动态变化。
使用奥林巴斯FV1000共聚焦显微镜，将来自主要癌症类型的双色细胞库培养在塑料上。通过共聚焦显微镜对细胞进行成像，以展示有丝分裂过程中的染色体和细胞质动态变化。
细胞核中GFP的表达能够观察到染色体行为，而同时细胞质中RFP的表达能够观察到有丝分裂过程中的细胞质行为。因此，在活的双色细胞中，可以实时高分辨率地观察到整个细胞的动态变化，在某些情况下还能观察到单个染色体。
在细胞核中表达H2B-GFP且在细胞质中表达RFP的双色癌细胞为观察亚细胞水平的细胞核和细胞质动态变化提供了独特的工具，包括有丝分裂过程中单个染色体的行为。双色细胞可用于实时评估在用各种药物治疗期间或在培养或体内选择细胞以增加或降低恶性程度时染色体的丢失或增加情况。双色细胞将成为发现和评估杀死癌细胞新策略的有用工具。
