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Alternative splicing of the FGF antisense gene: differential subcellular localization in human tissues and esophageal adenocarcinoma

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Abstract

Overexpression of FGF-2 is associated with tumor recurrence and reduced survival after surgical resection of esophageal cancer, and these risks are reduced in tumors co-expressing the FGF antisense (FGF-AS) RNA. The aim of this study was to characterize the expression of alternatively spliced FGF-AS transcripts and encoded nudix-motif proteins in normal human tissues and in esophageal adenocarcinoma, and to correlate their expression with clinicopathologic findings and outcome. Three alternatively spliced FGF-AS transcripts encoding GFG/NUDT6 isoforms with distinct N termini were detected in various human tissues including esophageal adenocarcinoma. Expression of each isoform as a fusion protein with enhanced green fluorescent protein revealed differential subcellular trafficking: hGFGa is localized to mitochondria by an N-terminal targeting sequence (MTS), whereas hGFGb and hGFGc were localized in the cytoplasm and nucleus. Mutation/deletion analysis confirmed that the predicted MTS was necessary and sufficient for mitochondrial compartmentalization. The predominant FGF-AS mRNA expressed in esophageal tumors was splice variant b. GFG immunoreactivity was detected in the cytoplasm of all esophageal adenocarcinomas and in 88% of tumor cell nuclei. Although we found a trend towards reduced disease-free survival in patients with FGF-2 overexpressing esophageal adenocarcinomas, significantly worse disease-free survival was noted among patients whose tumors did not also overexpress the FGF-AS b isoform (p = 0.03). Tetracycline-inducible FGF-AS b expression in stably transfected human Seg-1 esophageal adenocarcinoma cells resulted in a significant suppression of steady state FGF-2 mRNA content and cell proliferation. Our data implicate the FGF-AS b isoform in modulation of FGF-2 expression and clinical outcome in esophageal adenocarcinoma.

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Acknowledgements

We thank the Dalhousie Cancer Research and Training Program for postdoctoral fellowship support to SC Zhang. AG Casson and GA Porter were supported by a Clinical Research Scholarships from the Dalhousie University Faculty of Medicine. We also wish to thank Ms. R. Mi for excellent technical assistance and Dr. Mark Nachtigal for critical reading of the manuscript. This project was funded by grants from the Canadian Institutes of Health Research and the Cancer Research Society Inc.

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Authors and Affiliations

  1. Department of Physiology and Biophysics, Faculty of Medicine, Dalhousie University, 5850 College Street, Halifax, NS, B3H 1X5, Canada

    Shuo Cheng Zhang, Christie Barclay, Leigh Ann Alexander & Paul R. Murphy

  2. Department of Pathology, Faculty of Medicine, Dalhousie University, 5850 College Street, Halifax, NS, B3H 1X5, Canada

    Laurette Geldenhuys & Alan G. Casson

  3. Department of Surgery, Faculty of Medicine, Dalhousie University, 5850 College Street, Halifax, NS, B3H 1X5, Canada

    Geoffrey A. Porter & Alan G. Casson

  4. Department of Surgery, University of Saskatchewan, Royal University Hospital, Saskatoon, SK, S7N 0W8, Canada

    Alan G. Casson

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  1. Shuo Cheng Zhang
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  2. Christie Barclay
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  3. Leigh Ann Alexander
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  4. Laurette Geldenhuys
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Correspondence to Paul R. Murphy.

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Zhang, S.C., Barclay, C., Alexander, L.A. et al. Alternative splicing of the FGF antisense gene: differential subcellular localization in human tissues and esophageal adenocarcinoma. J Mol Med 85, 1215–1228 (2007). https://doi.org/10.1007/s00109-007-0219-9

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  • DOI: https://doi.org/10.1007/s00109-007-0219-9

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