title: The Use of Living Cancer Cells Expressing Green Fluorescent Protein in the Nucleus and Red Fluorescence Protein in the Cytoplasm for Real-time Confocal Imaging of Chromosome and Cytoplasmic Dynamics During Mitosis.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: 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.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Suetsugu
authors.affiliation: AntiCancer Inc., San Diego, CA, U.S.A.
authors.forename: Ping
authors.initials: P
authors.lastname: Jiang
authors.affiliation: AntiCancer Inc., San Diego, CA, U.S.A. AntiCancer Biotech (Beijing) Co., Ltd., Beijing, P.R. China.
authors.forename: Meng
authors.initials: M
authors.lastname: Yang
authors.affiliation: AntiCancer Inc., San Diego, CA, U.S.A. Department of Community Tramatology and Orthopaedics, Kanazawa University, Kanazawa, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Yamamoto
authors.affiliation: Department of Gastroenterology, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Hisataka
authors.initials: H
authors.lastname: Moriwaki
authors.affiliation: Department of Gastroenterology, Gifu University Graduate School of Medicine, Gifu, Japan.
authors.forename: Shigetoyo
authors.initials: S
authors.lastname: Saji
authors.affiliation: AntiCancer Inc., San Diego, CA, U.S.A. Department of Surgery, University of California, San Diego, CA, U.S.A. all@anticancer.com.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hoffman
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics / Q000473:pathology; D058948:Cell Tracking / Q000379:methods; D003593:Cytoplasm / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D008856:Microscopy, Fluorescence; D008938:Mitosis; D057054:Molecular Imaging; D009369:Neoplasms / Q000235:genetics / Q000473:pathology*; D000097573:Red Fluorescent Protein
chemical_list: D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: Cancer cells; GFP; RFP; chromosomes; cytoplasmic labeling; fusion; histone-H2B; mitosis; nuclear labeling; retrovirus