Skip to main content
SpringerLink
Log in

Immunocytochemical studies on the distribution pattern of daunomycin in rat gastrointestinal tract

  • Short Communication
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

The cancer drug daunomycin is used in treatment of leukemia but possesses severe side effects that involve the gastrointestinal tract. We therefore used a newly developed immunocytochemical procedure to determine the distribution of DM in the gastrointestinal tracts of rats after i.v. injection. Two hours after injection, DM was diffusely distributed in nuclei and most parts of the cytoplasm of intestinal epithelial cells. The cytoplasmic immunoreactivity for DM was most pronounced in small granules of the apical cytoplasm. Sixteen hours after injection, DM immunostaining was by and large absent in the villous epithelium but persisted in the intestinal crypts. In addition, staining was also detected in endothelial cells, scattered cells of the lamina propria and in smooth muscle cells. After 5 days, only little staining for DM remained. Similar findings were made in the colon. In the gastric mucosa, DM accumulation persisted at 16 h in some glandular cells but was lost from the surface epithelium. No staining was detected in saline-injected control rats. The distribution of DM accumulation correlated partially with the distribution of apoptotic cells as detected by the TUNEL procedure. Our results pinpoint that DM may exert prolonged effects on glandular and regenerative cells of the gastrointestinal tract—an observation that may explain the gastrointestinal toxicity of the drug. It seems possible that DM accumulation in surface epithelial cells is rapidly cleared through drug transporters.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Abbreviations

DM:

Daunomycin

ICC:

Immunocytochemistry

HRP:

Horseradish peroxidase

PBS:

Phosphate-buffered saline

RT:

Room temperature

TdT:

Terminal deoxynucleotidyl transferase

DAB:

3,3′-Diaminobenzidine

References

  • Adams JC (1981) Heavy metal intensification of DAB-based HRP reaction product. J Histochem Cytochem 29:775

    PubMed  Google Scholar 

  • Anikumar TV, Sarraf CE, Hunt T, Alison MR (1992) The nature of cytotoxic drug-induced cell death in murine intestinal crypts. Br J Cancer 65:552–558

    Google Scholar 

  • Bittl A, Nap M, Jager W, Cornillie F, Lang N (1995) Immunohistochemical detection of P-glycoprotein on frozen and paraffin-embedded tissue sections of normal and malignant tissues. Anticancer Res 15(3):1007–1014

    PubMed  Google Scholar 

  • Blum RH, Carter SK (1974) Adriamycin. A new anticancer drug with significant clinical activity. Ann Intern Med 80:249–259

    PubMed  Google Scholar 

  • Bristow MR (1982) Toxic cardiomyopathy due to doxorubicin. Hosp Pract 18:291–300

    Google Scholar 

  • Burke JF, Laucius JF, Brodovsky HS, Soriano RZ (1977) Doxorubicin hydrochloride-associated renal failure. Arch Intern Med 137:385–388

    Article  PubMed  Google Scholar 

  • Chabner BA, Ryan DP, Paz-Ares L, Carbonero RG, Calabresi P (2001) Antineoplastic agents. In: Hardman JG, Limbird LE, Gilman AG (eds) Goodman and Gilman’s the pharmacological basis of therapeutics, 10th edn. McGraw-Hill, New York, pp 1389

    Google Scholar 

  • Cherrington NJ, Hartley DP, Li N, Johnson DR, Klaassen CD (2002) Organ distribution of multidrug resistance protein 1, 2, and 3 (Mep 1,2, and 3) mRNA and hepatic induction of Mrp 3 by constitutive androstane receptor activators in rats. J Pharmacol Exp Ther 300:97–104

    Article  PubMed  Google Scholar 

  • Damavandi E, Hishikawa Y, Izumi S, Shin M, Koji T (2002) Involvement of Bax redistribution in the induction of germ cell apoptosis in neonatal mouse testis. Acta Histochem Cytochem 35:449–459

    Article  Google Scholar 

  • Dimarco A, Gaetani M, Dorigotti L, Soldati M, Bellini O (1963) Experimental studies of the antineoplastic activity of a new antibiotic daunomycin. Tumori 49:203–217

    PubMed  Google Scholar 

  • DuBuske LM (2005) The role of P-glycoprotein and organic anion-transporting polypeptides in drug interactions. Drug Saf 28:789–801

    Article  PubMed  Google Scholar 

  • Egorin MJ, Hildebrand RC, Cimino EF, Bachur NR (1974) Cytofluorescence localization of adriamycin and daunorubicin. Cancer Res 34:2243–2245

    PubMed  Google Scholar 

  • Ernest S, Bello-Reuss E (1995) Expression and function of P-glycoprotein in mouse kidney cell line. Am J Physiol 269:C323–C333

    PubMed  Google Scholar 

  • Fujiwara K, Takatsu H, Tsukamoto K (2005) Immunocytochemistry for drugs containing an aliphatic amino group in the molecule, anticancer antibiotic daunomycin as a model. J Histochem Cytochem 53:467–474

    Article  PubMed  Google Scholar 

  • Fujiwara K, Shin M, Hougaard MH, Larsson LI (2007) Distribution of anticancer antibiotic daunomycin in the rat heart an kidney revealed by immunocytochemistry using monoclonal antibodies. Histochem Cell Biol 127:69–77

    Article  PubMed  Google Scholar 

  • Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501

    Article  PubMed  Google Scholar 

  • Graham RC, Karnovsky MJ (1966) glomerular permeability. Ultrastructural cytochemitry by a new technique. J Histochem Cytochem 14:291–302

    PubMed  Google Scholar 

  • Hussain I, Kellett L, Affleck J, Shepherd J, Boyd R (2002) Expression and cellular distribution during development of the peptide transporter (PepT1) in the small intestinal epithelium of the rat. Cell Tissue Res 307(1):139–142

    Article  PubMed  Google Scholar 

  • Ijiri K, Potten CS (1987) Further studies on the response of intestinal crypt cells of different hierarchical status to eighteen different cytotoxic agents. Br J Cancer 55:113–123

    PubMed  Google Scholar 

  • Katsura T, Inui K (2003) Intestinal absorption of drugs mediated by drug transporters: mechanisms and regulation. Drug Metab Pharmacokinet 18(1):1–15

    Article  PubMed  Google Scholar 

  • Leu BL, Huang JD (1995) Inhibition of intestinal P-glycoprotein and effects on etoposide absorption. Cancer Chemother Pharmacol 35:432–436

    Article  PubMed  Google Scholar 

  • Ohara K, Shin M, Larsson LI, Fujiwara K (2007) Improved immunocytochemical detection of daunomycin. Histochem Cell Biol 127(6):603–608

    Article  PubMed  Google Scholar 

  • Pan Y, Wong EA, Bloomquist JR, Webb KE Jr (2001) Expression of a cloned ovine gastrointestinal peptide transporter (oPepT1) in Xenopus oocytes induces uptake of oligopeptides in vitro. J Nutr 131(4):1264–1270

    PubMed  Google Scholar 

  • Paschoud N, Bignell C, Reinhold HS (1985) A double fluorescence method for detecting doxorubicin distribution and vascular supply in the mouse kidney. J Histochem Cytochem 33:73–76

    PubMed  Google Scholar 

  • Platel D, Bonorone-Adele S, Robert J (2001) role of daunorubicinol in daunomycin-induced cardiotoxicity as evaluated with the model of isolated perfused rat heart. Pharmacol toxicol 88:250–254

    Article  PubMed  Google Scholar 

  • Rutherford A, Willingtham MC (1993) Ultrastructural localization of daunomycin in multidrug-resistant culture cells with modulation of the multidrug transpoter. J Histochem Cytochem 41:1573–1577

    PubMed  Google Scholar 

  • Schinkel AH, Mayer U, Wagenaar E, Mol CA, van Deemter L, Smit JJ, van der Valk MA, Voordouw AC, Spits H, van Tellingen O, Zijlmans JM, Fibbe WE, Borst P (1997) Normal viability and altered pharmacokinetics in mice lacking mdr1-type (drug-transporting) P-glycoproteins. Proc Natl Acad Sci USA 94:4028–4033

    Article  PubMed  Google Scholar 

  • Takahashi S, Bachur NR (1975) Daunorubicin metabolites in human urine. J Pharmacol Exp Ther 195:41–49

    Google Scholar 

  • Tanigawara Y (2000) Role of P-glycoprotein in drug disposition. The Drug Monit 22:137–140

    Article  Google Scholar 

  • Thakkar NS, Potten CS (1993) Inhibition of doxorubicin-induced apoptosis in vivo by 2-deoxy-D-glucose. Cancer Res 53:2057–2060

    PubMed  Google Scholar 

  • Thakkar NS, Potten CS (1992) Abrogation of adriamycin toxicity in vivo by cycloheximide. Biochem Pharmacol 43:1683–1691

    Article  PubMed  Google Scholar 

  • Thiebaut F, Tsuruo T, Hamada H, Gottesman MM, Pastan I, Willingham MC (1987) Cellular localization of the multidrug-resistance gene product P-glycoprotein in normal human tissues. Proc Natl Acad Sci USA 84:7735–7738

    Article  PubMed  Google Scholar 

  • Tsuji A (2006) Impact of transporter-mediated drug absorption, distribution, elimination and drug interactions in antimicrobial chemotherapy. J Infect Chemother 12:241–250

    Article  PubMed  Google Scholar 

  • van Asperen J, van Tellingen O, van der Valk MA, Rozenhart M, Beijnen JH (1998) Enhanced oral absorption and decreased elimination of paclitaxel in mice cotreated with cyclosporin A. Clin Cancer Res 4:2293–2297

    PubMed  Google Scholar 

  • Willingham MC, Cornwell MM, Cardarelli CO, Gottesman MM, Pastan I (1986) Single cell analysis of daunomycin uptake and efflux in multidrug-resistant and sensitive KB cells: Effects of verapamil and other drugs. Cancer Res 46:5941–5946

    PubMed  Google Scholar 

  • Zagotto G, Gatto B, Moro S, Sissi C, palumbo M (2001) Anthracyclines: recent developments in their separation and quantification. J Chromatogr B Biomed Sci Appl 764:161–171

    Article  PubMed  Google Scholar 

  • Zhang J, Clark JR, Herman EH, Ferrans VJ (1996) Doxorubicin-induced apoptosis in spontaneously hypertensive rats: differential effects in heart, kidney and intestine, and inhibition by ICRF-187. J Mol Cell Cardiol 28:1931–1943

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We are grateful to Mariko Takauye and Yuuta Tamamura for their technical assistance throughout this study.

Author information

Authors and Affiliations

  1. Department of Applied Life Science, Faculty of Biotechnology and Life Science, Sojo University, Ikeda 4-22-1, Kumamoto, 860-0082, Japan

    Koji Ohara, Masashi Shin & Kunio Fujiwara

  2. Laboratory of Pharmacology, Department of Pharmaceutical Sciences, Faculty of Pharmaceutical Sciences, Hiroshima International University, Hirokoshingai 5-1-1, Kure, Hiroshima, 737-0112, Japan

    Hiromichi Nakamuta

  3. Department of Clinical Biochemistry, Statens Serum Institute, Copenhagen, Denmark

    David M. Hougaard

  4. Department of Cell Biology IBHV, Faculty of Life Sciences, University of Copenhagen, Gronnegaardsvej 7, 1870, Frederiksberg C, Denmark

    Lars-Inge Larsson

Authors
  1. Koji Ohara
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Masashi Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiromichi Nakamuta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Lars-Inge Larsson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. David M. Hougaard
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kunio Fujiwara
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kunio Fujiwara.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ohara, K., Shin, M., Nakamuta, H. et al. Immunocytochemical studies on the distribution pattern of daunomycin in rat gastrointestinal tract. Histochem Cell Biol 128, 285–290 (2007). https://doi.org/10.1007/s00418-007-0314-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-007-0314-6

Keywords

Search

Navigation