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Detection of Cyclin B1 Expression in G₁-Phase Cancer Cell Lines and Cancer Tissues by Postsorting Western Blot Analysis

¹ Tongji Cancer Institute and Center for Molecular Medicine, Department of Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, People’s Republic of China, and ² Mary Babb Randolph Cancer Center and Department of Microbiology, Immunology and Cell Biology, West Virginia University School of Medicine and Robert C. Byrd Health Sciences Center, Morgantown, West Virginia

Protein complex of cyclin B1 and cyclin-dependent protein kinase 1 induces phosphorylation of key substrates that mediate cell cycle transition during the G₂-M phase. It is believed that cyclin B1 accumulates in the S phase of the cell cycle and reaches the maximal level at mitosis but is absent in G₁-phase cells. In the present study, we demonstrated that cyclin B1 was expressed in the arrested G₁-phase MOLT-4 lymphocyte leukemia cells and in G₁ phase T-7 transitional tumor cells, as determined by flow cytometry. In addition, we showed that cyclin B1 was detected in the G₁ phase in breast cancer cells from patient tissues and in lymphocytes from patients with leukemia. These findings were confirmed for the first time by postsorting Western blot analysis and by confocal microscopy. Furthermore, by using postsorting Western blotting, we found that cyclin B1 was expressed in different time-window sections of the G₁ phase under different conditions. For the asynchronously growing T-7 cells, cyclin B1 was detected in early G₁ phase, whereas in MOLT-4 cells arrested in G₁-S phase, cyclin B1 was mainly detected in late G₁ phase. We propose that the cyclin B1 expressed in the G₁ phase may differ from that expressed in the G₂-M phase, and that this unscheduled type of cyclin B1 may play an important role in tumorigenesis and apoptosis.

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