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Nuclear co-expression of p21 and p27 induced effective cell-cycle arrest in T24 cells treated with BCG

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Abstract

A proposed mechanism underlying the effect of bacillus Calmette–Guérin (BCG) treatment for bladder cancer cells is as follows: BCG-induced crosslinking of cell-surface receptors results in the activation of signaling cascades, including cell-cycle regulators. However, the clinical significance of cell-cycle regulators such as p21 and p27 is controversial. Here we investigated the relationship between BCG exposure and p21 and p27. We used confocal laser microscopy to examine the expression levels of pKi67, p21 and p27 in T24 cells (derived from human urothelial carcinoma) exposed six times to BCG. We performed dual immunofluorescence staining methods for p21 and p27 and observed the localization of nuclear and cytoplasm expressions. We investigated the priority of p27 over p21 regarding nuclear expression by using p27 Stealth RNAi™ (p27-siRNA). With 2-h BCG exposure, the nuclear-expression level of p21 and p27 was highest, while pKi67 was lowest. The percentage of double nuclear-expression of p21 and p27 in BCG cells was significantly higher than that in control cells during the 1st to 6th exposure (P < 0.05), and the expression of pKi67 showed the opposite of this pattern. Approximately 10% of the nuclear p21 was independent of p27, whereas the cytoplasmic p21 was dependent on p27. Our results suggested that the nuclear co-expression of p21 and p27 caused effective cell-cycle arrest, and thus the evaluation of the nuclear co-expression of p21 and p27 might help determine the effectiveness of BCG treatment.

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Acknowledgements

This study was supported by JSPS KAKENHI Grant Nos. #JP25460459 and #JP17K08744.

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Authors and Affiliations

  1. Department of Health Sciences, Faculty of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka City, 812-8582, Japan

    Sumiko Watanabe, Hitoshi Katsuta, Setsuo Sugishima & Tsunehisa Kaku

  2. Department of Health Sciences, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka City, 812-8582, Japan

    Shota Yamaguchi, Naoto Fujii & Natsuki Eguchi

  3. Department of Obstetrics and Gynecology, Takagi Hospital, 141-11, Sakemi, Okawa City, Fukuoka, 831-0016, Japan

    Tsuyoshi Iwasaka

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  1. Sumiko Watanabe
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  2. Shota Yamaguchi
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  3. Naoto Fujii
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  4. Natsuki Eguchi
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  5. Hitoshi Katsuta
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Correspondence to Sumiko Watanabe.

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Watanabe, S., Yamaguchi, S., Fujii, N. et al. Nuclear co-expression of p21 and p27 induced effective cell-cycle arrest in T24 cells treated with BCG. Cytotechnology 71, 219–229 (2019). https://doi.org/10.1007/s10616-018-0278-5

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  • DOI: https://doi.org/10.1007/s10616-018-0278-5

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