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Impact of single-nucleotide polymorphisms at the TP53-binding and responsive promoter region of BCL2 gene in modulating the phenotypic variability of LGMD2C patients

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Abstract

Apoptosis of skeletal muscle fibers is a well-known event occurring in patients suffering from muscular dystrophies. In this study, we hypothesized that functional polymorphisms in genes involved in the mitochondrial apoptotic pathway might modulate the apoptotic capacity underlying the muscle loss and contributing to intrafamilial and interfamilial variable phenotypes in LGMD2C (Limb Girdle Muscular Dystrophy type 2C) patients sharing the same c.521delT mutation in SGCG gene. Detection of apoptosis was confirmed on muscle biopsies taken from LGMD2C patients using the TUNEL method. We genotyped then ten potentially functional SNPs in TP53, BCL-2 and BAX genes involved in the mitochondrial apoptotic pathway. Potential genotype-dependent Bcl-2 and p53 protein expressed in skeletal muscle was investigated using western blot and ELISA assays. The result showed that muscle cells carrying the TP53-R72R and TP53-16 bp del/del genotypes displayed an increased p53 level which could be more effective in inducing apoptosis by activation of the pro-apoptotic gene expression. In addition, the BCL2-938 AA genotype was associated with increased Bcl-2 protein expression in muscle from LGMD2C patients compared to −938CC genotype, while there was no evidence of significant difference in the BAX haplotype. Our findings suggest that increased Bcl-2 protein expression may counteract pro-apoptotic pathways and thus reduce the muscle loss. To the best of our knowledge, this is a pioneer study evaluating the role of apoptotic BCL-2 and TP53 genes in contributing to the phenotypic manifestation of c.521delT mutation in LGMD2C patients. Larger studies are needed to validate these findings.

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Acknowledgments

We are grateful to the family members for their invaluable cooperation. We are grateful to Pr. Raja Mokdad Gargouri from Centre of Biotechnology of Sfax, for providing us the p53 antibody. We also acknowledge Mr. Jamil JAOUA, founder and former head of the English Unit at the Sfax Faculty of Science, Tunisia, for proofreading this paper.

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Authors and Affiliations

  1. Laboratoire de Génétique Moléculaire Humaine, Faculté de Médecine de Sfax, Université de Sfax, Avenue Magida Boulila, 3029, Sfax, Tunisia

    Ikhlass Hadj Salem, Nacim Louhichi & Faiza Fakhfakh

  2. Service de Neurologie Pédiatrique, CHU Hédi Chaker de Sfax, Sfax, Tunisia

    Fatma Kamoun & Chahnez Triki

  3. Service d’Orthopédie et Traumatologie, CHU Habib-Bourguiba de Sfax, Sfax, Tunisia

    Moez Trigui

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  1. Ikhlass Hadj Salem
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  2. Fatma Kamoun
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  3. Nacim Louhichi
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  4. Moez Trigui
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  5. Chahnez Triki
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  6. Faiza Fakhfakh
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Correspondence to Ikhlass Hadj Salem or Faiza Fakhfakh.

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Hadj Salem, I., Kamoun, F., Louhichi, N. et al. Impact of single-nucleotide polymorphisms at the TP53-binding and responsive promoter region of BCL2 gene in modulating the phenotypic variability of LGMD2C patients. Mol Biol Rep 39, 7479–7486 (2012). https://doi.org/10.1007/s11033-012-1581-4

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  • DOI: https://doi.org/10.1007/s11033-012-1581-4

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