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Reports

Imaging of Nucleolar Dynamics During the Cell Cycle of Cancer Cells in Live Mice

Kensuke Yamauchi, Meng Yang, Katsuhiro Hayashi, Ping Jiang, Norio Yamamoto, Hiroyuki Tsuchiya, Katsuro Tomita, A.R. Moossa, Michael Bouvet and Robert M. Hoffman

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The synthesis and assembly of ribosomal subunits take place in the nucleolus. The nucleolus forms in the nucleus around the repeated ribosomal gene clusters and undergoes cyclic changes during the cell cycle. Although the nucleolus is easily visualized by light microscopy of cells in vitro, the nucleolus has not been imaged in cells in vivo. We report here development of a mouse model to visualize the nucleolus cycle of cancer cells in live mice. HT-1080 human fibrosarcoma cells were labeled in the nucleus with histone H2B-GFP and with retroviral RFP in the cytoplasm. The nucleolus was visualized by contrast to the fluorescence of GFP expressed in the nucleus. HT-1080 dual-color cells were seeded on the surface of a skin-flap of nude mice. The inside surface of the skin-flap was directly imaged with a laser scanning microscope 24 hours after seeding. The nucleoli of the cancer cells were clearly imaged in real-time. The appearance of the nucleoli changed dramatically during the cell cycle. During mitosis, the nucleolus disappeared. After mitosis, the nucleoli decreased in number and increased in size. The nucleolus appears to have a major role in cell cycle regulation effected at least in part by sequestering proteins which affect cell cycle progression. Nucleolar imaging could be used for more precise determination of cancer-cell position in the cell cycle in vivo.

Authors

Kensuke Yamauchi

AntiCancer, Inc.; San Diego, CA

Meng Yang

AntiCancer, Inc.; San Diego, CA

Katsuhiro Hayashi

AntiCancer, Inc.; San Diego, CA

Ping Jiang

AntiCancer, Inc.; San Diego, CA

Norio Yamamoto

Kanazawa University; Kanazawa, Ishikawa, Japan

Hiroyuki Tsuchiya

Kanazawa University; Kanazawa, Ishikawa, Japan

Katsuro Tomita

Kanazawa University; Kanazawa, Ishikawa, Japan

A.R. Moossa

University of California; San Diego, CA

Michael Bouvet

University of California; San Diego, CA

Robert M. Hoffman

AntiCancer, Inc.; San Diego, CA

Purchase article for $19

Subscribe to this journal for $129/year