Effects of hyaluronidase on doxorubicin penetration into squamous carcinoma multicellular tumor spheroids and its cell lethality (original) (raw)

Abstract

Doxorubicin is an anticancer agent widely used in the treatment of human cancer. The major limitation of this drug governing the cell-killing effect appears to be its poor penetration into a tumor mass. We have studied the effects of hyaluronidase on the penetration and cell-killing effect of doxorubicin using multicellular tumor spheroids (MTS). MTS approximately 500 μm in diameter were produced by a liquid-overlay culture technique from PC-10 lung and HEp-2 laryngeal squamous carcinoma cell lines. Cells in MTS and monolayer were exposed to hyaluronidase for various lengths of time; this was followed by a 1-h resting interval and a subsequent 1-h exposure to doxorubicin. MTS and monolayer cells were then trypsinized to a single-cell suspension and subjected to clonogenic assay. Hyaluronidase at a concentration of 25 U/ml or 250 U/ml was nontoxic to the monolayer cells. For PC-10 MTS, pretreatment with 25 U/ml hyaluronidase for 24 h and 72 h resulted in approximately 20% increases in Doxorubicin cell killing at the median (IC50) dose as compared to doxorubicin alone. HEp-2 MTS were more sensitive to the hyaluronidase pretreatment. Thus, a 1-h exposure to the enzyme produced a 40% increase in doxorubicin-induced cell death at the IC50 dose. A fluorescence microscopic study revealed that a 1-h exposure of MTS to doxorubicin produced doxorubicin fluorescence only in the one or two outer layers of MTS. When MTS were pretreated with hyaluronidase, there was enhanced penetration of doxorubicin fluorescence into the MTS core. Hyaluronidase-induced enhancement of Doxorubicin penetration and its cell-killing effect is dependent on the exposure time and tumor cell origin. These data suggest that anecdotal reports of hyaluronidase-enhanced activity of preclinical chemotherapy deserve a controlled trial.

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Abbreviations

MTS:

multicellular tumor spheroids

References

• Baumgartner G (1987) Hyaluronidase in der Therapie maligner Erkrankungen. Wien Klin Wochenschr 15 [Suppl 175]:99
  Google Scholar
• Baumgartner G, Fortelny A, Zänker KS, Kroczek R (1988) Phase I study in chemoresistant loco-regional malignant disease with hyaluronidase. Reg Cancer Treat 1:55–58
  Google Scholar
• Bender RA, Bleyer WA, Friesby SA, Oliverio VT (1975) Alteration of methotrexate uptake in human leukemia cells by other agents. Cancer Res 35:1305–1308
  PubMed Google Scholar
• Erlichmann C, Vidgen D (1984) Cytotoxicity of adriamycin in MGHU1 cells grown as monolayer cultures, sphereroids, and xenografts in immune-deprived mice. Cancer Res 44:5369–5375
  PubMed Google Scholar
• Folkman J (1986) How is blood vessel growth regulated in normal and neoplastic tissue? G.H.A. Clowes Memorial Award Lecture. Cancer Res 46:467–473
  PubMed Google Scholar
• Horn Y, Eidelman A, Walach N, Waron M, Barak F (1985) Intravesical chemotherapy of superficial bladder turmors in a controlled trial with cis-platinum versus cis-platinum plus hyaluronidase. J Surg Oncol 28:304–307
  PubMed Google Scholar
• Inoue S, Ohnuma T, Takaoka K, Suzuki Y, Kaneko M, Safirstein R, Holland J (1987) Effects of doxorubicin and cisplatin in multicellular tumor spheroids from human lung cancer. Cancer Drug deliv 4:213–224
  PubMed Google Scholar
• Kinjo M, Oka K, Naito S, Kohga S, Tanaka K, Oboshi S, Hayata Y, Yasumoto K (1979) Thromboplastic and fibrinolytic activities of cultured human cancer lines. Br J Cancer 39:15–23
  PubMed Google Scholar
• Kohno N, Ohnuma T, Kaneko M, Holland JF (1988) Interactions of doxorubicin and cisplatin in squamous carcinoma in culture. Br J Cancer 58:330–334
  PubMed Google Scholar
• Kuroki T (1974) Colony formation of mamalian cells of agar plates and its application to Lederberg's replica plating. Exp Cell Res 80:55–62
  Google Scholar
• Maier U, Baumgartner G (1989) Metaphylactic effect of mitomycin C with and without hyaluronidase after transurethral resection of bladder cancer: randomized trial. J Urol 141:529–530
  PubMed Google Scholar
• Sutherland RM, Durand RE (1984) Growth and cellular characteristics of multicell spheroids. Recent Results Cancer Res 95:24–48
  PubMed Google Scholar
• Thomlison RH, Gray LH (1955) The histological structure of some human lung cancer and the possible implications for radiotherapy. Br J Cancer 9:539–549
  PubMed Google Scholar
• Toburen D (1981) Differential survival of cell subpopulations in spheroids after treatment with bleomycin. J Cancer Res Clin Oncol 102:189–193
  PubMed Google Scholar
• Warren RD, Nichols AP, Bender RA (1977) The effect of vincristine on methotrexate uptake and inhibition of DNA synthesis by human lymphoblastoid cells. Cancer Res 37:2993–2997
  PubMed Google Scholar
• West GW, Weichselbaum R, Little JB (1980) Limited penetration of methotrexate into human osteosarcoma spheroids as a proposed model for solid tumor resistance to adjuvant Chemotherapy. Cancer Res 40:3665–3668
  PubMed Google Scholar
• Yuhas JM, Li AP, Martinez AD, Ladman AJ (1977) A simplified method for production and growth of multicellular tumor spheroids. Cancer Res 37:3639–3643
  PubMed Google Scholar

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Authors and Affiliations

1. Department of Otolaryngology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, 113, Tokyo, Japan
   Naoyuki Kohno
2. Department of Neoplastic Disease, Mount Sinai School of Medicine, New York, USA
   Takao Ohnuma
3. St.-Johanns-Vorstadt 38, 4056, Basel, Switzerland
   Peter Truog

Authors

1. Naoyuki Kohno
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2. Takao Ohnuma
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3. Peter Truog
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Additional information

This work was supported in part by the Chemotherapy Foundation, Inc., New York, NY; by the T. J. Martell Foundation for Leukemia and Cancer Research, New York, NY and by the Hayashibara Biochemical Laboratories, Inc., Okayama, Japan

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Kohno, N., Ohnuma, T. & Truog, P. Effects of hyaluronidase on doxorubicin penetration into squamous carcinoma multicellular tumor spheroids and its cell lethality.J Cancer Res Clin Oncol 120, 293–297 (1994). https://doi.org/10.1007/BF01236386

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• Received: 01 June 1993
• Accepted: 21 September 1993
• Issue Date: May 1994
• DOI: https://doi.org/10.1007/BF01236386

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