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Effect of transforming growth factor-alpha on enterocyte apoptosis is correlated with EGF receptor expression along the villus-crypt axis during methotrexate-induced intestinal mucositis in a rat

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Abstract

The purpose of the present study was to evaluate the effect of transforming-growth factor-alpha (TGF-α) on enterocyte apoptosis following methotrexate (MTX) induced intestinal mucositis in a rat and in Caco-2 cells. Non-pretreated and pretreated with MTX Caco-2 cells were incubated with increasing concentrations of TGF-α. Cell apoptosis was determined by FACS cytometry. Adult rats were divided into four groups: Control, Control-TGF-α, MTX, and MTX- TGF-α rats. Three days later rats were sacrificed. Enterocyte apoptosis were measured at sacrifice. RT-PCR and Western Blotting was used to determine the level of Bax and Bcl-2 mRNA and protein. Real time PCR was used to measure epidermal growth factor receptor (EGFr) expression along the villus-crypt axis. The in vitro experiment has shown that treatment with TGF-α of Caco-2 cells results in a significant inhibition of cell apoptosis in a dose-dependent manner. In vivo experiment, a decreased levels of apoptosis in MTX- TGF-α rats corresponded with the decrease in Bax and with the increase in Bcl-2 at both mRNA and protein levels. The inhibiting effect of TGF-α on enterocyte apoptosis was strongly correlated with EGFr expression along the villus-crypt axis. In conclusion, treatment with TGF-α inhibits enterocyte apoptosis following MTX- injury in the rat.

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Acknowledgments

This work was supported by a research grant from VFK Krebsforschung gGmbH in Berlin, Germany.

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Authors and Affiliations

  1. Laboratory of intestinal adaptation and recovery, The Bruce Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel

    Igor Sukhotnik, Dan Shteinberg, Shani Ben Lulu & Yulia Bashenko

  2. Department of Pediatric Surgery, Bnai Zion Medical Center, 47 Golomb St., P. O. Box 4940, 31048, Haifa, Israel

    Igor Sukhotnik, Jorge G. Mogilner & Ron Shaoul

  3. Department of Pediatric Surgery, Hannover Medical School, Hannover, Germany

    Benno M. Ure

  4. Section of Pediatric Surgery C.S, Mott Children’s Hospital and University of Michigan Medical School, Ann Arbor, MI, USA

    Arnold G. Coran

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  1. Igor Sukhotnik
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  2. Dan Shteinberg
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Correspondence to Igor Sukhotnik.

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Sukhotnik, I., Shteinberg, D., Ben Lulu, S. et al. Effect of transforming growth factor-alpha on enterocyte apoptosis is correlated with EGF receptor expression along the villus-crypt axis during methotrexate-induced intestinal mucositis in a rat. Apoptosis 13, 1344–1355 (2008). https://doi.org/10.1007/s10495-008-0258-x

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