Imaging of nucleolar dynamics during the cell cycle of cancer cells in live mice.
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. Nucleolar imaging could be used for more precise determination of cancer-cell position in the cell cycle in vivo.
Yamauchi
Kensuke
K
AntiCancer, Inc., San Diego, California 92111-3604, USA.
Yang
Meng
M
Hayashi
Katsuhiro
K
Jiang
Ping
P
Yamamoto
Norio
N
Tsuchiya
Hiroyuki
H
Tomita
Katsuro
K
Moossa
A R
AR
Bouvet
Michael
M
Hoffman
Robert M
RM
D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000737:chemistry* / Q000502:physiology*; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal / Q000379:methods