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High-intensity focused ultrasound induced apoptosis with caspase 3, 8, and 9/6 activation in rat hepatoma

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Abstract

Purpose

The purpose of the present study is to investigate anticancer efficacy and apoptosis confirmed by caspase under several exposure conditions of high-intensity focused ultrasound (HIFU).

Materials and methods

Twenty-five rats with KDH-8 hepatoma were treated by HIFU at several acoustic energies to evaluate treatment efficacy. Apoptosis was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) and Hoechst 33258 staining, and caspase 3, 8, and 9/6 activity was respectively assayed.

Results

The KDH-8 subcutaneous tumors were reduced by HIFU, and these rats survived longer than the nontreatment rats (P < 0.01). The minimal threshold of HIFU energy was 30 W × 1.0 s for tumor control and long-term survival. The tumors exposed to HIFU exhibited marked apoptotic features under conditions of less than 10 W × 1.0 s. In cultured KDH-8 cells, apoptosis was caused at less than 30 W × 1.0 s (P < 0.01), and more was induced as the energy went down. Caspase 3, 8, and 9/6 were more activated at low energy under 10 W × 1.0 s (P < 0.01), and caspase 8, which is death receptor dependent, was significantly more activated than caspase 9/6, which is mitochondria dependent (P < 0.01).

Conclusion

HIFU-induced apoptosis in vivo and in vitro is one of the mechanisms for tumor control and is mediated by caspase 3, 8, and 9/6. The significantly greater activation of caspase 8 than of caspase 9/6 suggests that the apoptosis pathway induced by HIFU might be more mitochondria dependent than death receptor dependent. However, further examination will be needed.

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Abbreviations

HIFU:

High-intensity focused ultrasound

SPTA:

Spatial peak temporal average

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling

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Authors and Affiliations

  1. Department of Radiology, School of Medicine, Sapporo Medical University, Minami-1, Nishi-16, Chuo-ku, Sapporo, 060-8543, Japan

    Naoki Hirokawa, Kazumitsu Koito, Takeshi Ichimura, Masakazu Hori, Taishi Satoh & Masato Hareyama

  2. Department of Biomolecular Function, Graduate School of Medicine, Yamagata University, Yamagata, 990-9585, Japan

    Futoshi Okada

  3. Laboratory of Biochemical Instrumentation & Measurements Research Associate, Graduate School of Engineering, Hokkaido University, Sapporo, 060-8628, Japan

    Nobuki Kudo & Katsuyuki Yamamoto

  4. Toshiba Medical Systems Corporation, Ohtawara, 324-8550, Japan

    Katsuhiko Fujimoto

  5. Division of Clinical Laboratory and Transfusion Medicine, Hokkaido University Hospital, Sapporo, 060-8628, Japan

    Mutsumi Nishida

Authors
  1. Naoki Hirokawa
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  2. Kazumitsu Koito
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  3. Futoshi Okada
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  4. Nobuki Kudo
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  5. Katsuyuki Yamamoto
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  6. Katsuhiko Fujimoto
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  7. Mutsumi Nishida
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  8. Takeshi Ichimura
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  9. Masakazu Hori
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  10. Taishi Satoh
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  11. Masato Hareyama
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Correspondence to Naoki Hirokawa.

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Hirokawa, N., Koito, K., Okada, F. et al. High-intensity focused ultrasound induced apoptosis with caspase 3, 8, and 9/6 activation in rat hepatoma. J Med Ultrasonics 36, 177–185 (2009). https://doi.org/10.1007/s10396-009-0234-2

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  • DOI: https://doi.org/10.1007/s10396-009-0234-2

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