Gülsevinç AKSOY, Ümit LÜLEYAP, Gülşah EVYAPAN, Perçin PAZARCI, Davut ALPTEKİN, Ayfer PAZARBAŞI, Mehmet Bertan YILMAZ

Sh-Sy5y hücre hattında sodyum bütiratın bazı alternatif kırpılma genleri ve BACE1 izoformları üzeindeki etkisi

Amaç: Bu çalışmanın amacı, bir Histon Deasetilaz (HDAC) inhibitörü olan Sodyum Bütirat’ın(NaB) BACE1/501, BACE1/457 ve BACE1/432 gibi BACE1 izoformları ve hnRNP H, U2AF35, U2AF65, SRSF1, SRSF2, SRSF5 ve SRSF6 alternatif kırpılma faktörleri üzerindeki etkisini araştırmaktır. Gereç ve Yöntem: Bu çalışmada, SH-SY5Y hücreleri 1 mM ve 5 mM Na Bile muamele edilmiştir. Daha sonra BACE1 izoformları ve alternative kırpılma genlerinin ekspresyonlarındaki değişimleri değerlendirmek için Real-Time PCR yöntemi kullanılmıştır. Bulgular: 5 mM NaB konsantrasyonunda SRSF6, SRSF1, SRSF2, SRSF5 ve U2AF65 genlerinin ekspresyonlarında %35’ten %80’e kadar görece bir azalma olduğu belirlenmiştir. Ayrıca 5 Mm NaB BACE1/501, BACE1/457 ve BACE1/432 izoformlarının ekspresyonunu artırmıştır. NaB konsantrasyonu arttıkça, SRSF5 geninin ekspresyonu diğerlerinden daha fazla azalmıştır. Sonuç: Bulgularımızdan yola çıkılarak, U2AF65 ve SRSF6 genlerinin BACE1/457 ve BACE1/432 izoformlarının artışına diğer genlerden daha fazla katkı sağladığı sonucuna varılmıştır.

Effect of sodium butyrate on some alternative splicing genes and BACE1 isoforms in Sh-Sy5y cell line

Purpose: The aim of this study is to investigate effect of sodium butyrate (NaB), which is a Histon Deacetylase (HDAC) inhibitor, on BACE1 isoforms BACE1/501, BACE1/457 and BACE1/432 and alternative splicing factors including hnRNP H, U2AF35, U2AF65, SRSF1, SRSF2, SRSF5 and SRSF6. Materials and Methods: In this study, SH-SY5Y cells were treated with of 1 mM and 5 mM NaB. Then Real-Time PCR method was performed to evaluate the change in expression of alternative splicing genes and BACE1 isoforms. Results: At 5 mM NaB concentration, a relative decrease of 35% to 80% in expression of the SRSF6, SRSF1, SRSF2, SRSF5 and U2AF65 genes were detected. Morever 5 mM NaB concentration increased expression of BACE1/501, BACE1457 and BACE1/432 isoforms. As NaB concentration increase, expression of SRSF6 mRNA decreased to a lower extent than others. Conclusion: According to our results, it can be concluded that contribution of the U2AF65 and SRSF6 genes to the increase of BACE1/457 and BACE1/432 expression is higher than the other genes.

Keywords:

Alzheimer’s disease, alternative splicing, sodium butyrate,

PDF

___

Marques SC et al. Alzheimer's disease: the quest to understand complexity. J Alzheimers Dis. 2010;21:373-383.
Sadigh-Eteghad S et al. Amyloid-beta: a crucial factor in Alzheimer's disease. Med Princ Pract. 2015;24:1-10.
Cole SL, Vassar R. The Alzheimer's disease beta-secretase enzyme. BACE1. Mol Neurodegener. 2007;2:22.
Love JE et al. Alternative Splicing in Alzheimer's Disease. J Parkinsons Dis Alzheimers Dis. 2015;2.
Wang Y et al. Mechanism of alternative splicing and its regulation. Biomed Rep. 2015;3(2): 152–158.
Vassar R, Cole S. The Basic Biology of BACE1: A Key Therapeutic Target for Alzheimers Disease. Curr Genomics. 2007;8:509-530.
Holsinger RMD et al. Splice variants of the Alzheimer’s disease beta-secretase, BACE1. Neurogenetics. 2012;14:1-9.
Kornblihtt AR et al. Alternative splicing: a pivotal step between eukaryotic transcription and translation. Nature Reviews Molecular Cell Biology. 2013;14:153-165.
Hacht Av et al. Identification and characterization of RNA guanine-quadruplex binding proteins. Nucleic Acids Res. 2014;42:6630-6644.
Fisette J-F et al. A G-Rich element forms a G-quadruplex and regulates BACE1 mRNA alternative splicing. J Neurochem. 2012;121:763-773.
Mowrer KR, Wolfe MS. Identification of acis-acting element involved in the regulation of BACE1 mRNA alternative splicing. J Neurochem. 2009;109:1008-1016.
Rahman MA et al. SRSF1 and hnRNP H antagonistically regulate splicing of COLQ exon 16 in a congenital myasthenic syndrome. Sci Rep. 2015;5.
Bayat S et al. HDACis (class I), cancer stem cell, and phytochemicals: Cancer therapy and prevention implications. Biomed Pharmacother. 2018;97:1445-1453.
Bonfils C et al. Pharmacological inhibition of histone deacetylases for the treatment of cancer, neurodegenerative disorders and inflammatory diseases. Expert Opinion on Drug Discovery. 2008;3:1041-1065.
Xuan et al. Valproic acid alleviates memory deficits and attenuates amyloid-β deposition in transgenic mouse model of Alzheimer's disease. Mol Neurobiol. 2015;51(1):300-12.
Wang L et al. Sodium Butyrate Induces Human Colon Carcinoma HT-29 Cell Apoptosis through a Mitochondrial Pathway. J Int Med Res. 2009;37:803-811.
Soldatenkov VA et al. Sodium butyrate induces apoptosis and accumulation of ubiquitinated proteins in human breast carcinoma cells. Cell Death Differ. 1998;5:307-312.
Hnilicová J et al. Histone Deacetylase Activity Modulates Alternative Splicing. PLoS One. 2011;6:e16727.
Natoni F et al. Sodium butyrate sensitises human pancreatic cancer cells to both the intrinsic and the extrinsic apoptotic pathways. Biochim Biophys Acta. 2005;1745:318-329.
Favaloro B et al.: Role of apoptosis in disease. Aging (Albany NY). 2012;4:330-349.
Piotrowska H, Jagodzinski PP. Trichostatin A, sodium butyrate, and 5-aza-2'-deoxycytidine alter the expression of glucocorticoid receptor alpha and beta isoforms in Hut-78 T- and Raji B-lymphoma cell lines. Biomed Pharmacother. 2007;61:451-454.
Govindarajan N et al. Sodium Butyrate Improves Memory Function in an Alzheimer's Disease Mouse Model When Administered at an Advanced Stage of Disease Progression. J Alzheimers Dis. 2011;26:187-197.
Cho S et al. Splicing inhibition of U2AF65leads to alternative exon skipping. Proceedings of the National Academy of Sciences. 2015;112:9926-9931.
Agrawal AA et al. Structure-guided U2AF(65) variant improves recognition and splicing of a defective pre-mRNA. Proc Natl Acad Sci USA. 2014;111:17420-17425.
Zhang J et al. Disease-associated mutation in SRSF2 misregulates splicing by altering RNA-binding affinities. Proc Natl Acad Sci USA. 2015;112:E4726-E4734.
Deschenes M, Chabot B. The emerging role of alternative splicing in senescence and aging. Aging Cell. 2017;16:918-933.
Shepard PJ, Hertel KJ. The SR protein family. Genome Biol. 2009;10:242.
Gipson TA et al. Aberrantly spliced HTT, a new player in Huntington's disease pathogenesis. RNA Biol. 2013;10:1647-1652.
Ceccacci E, Minucci S. Inhibition of histone deaceylases in cancer therapy: lessons from leukaemia. Br J Cancer. 2016; 114:605-611.
Davie JR. Inhibition of deacetylase activity by butyrate. J Nutr. 2003;132:2485S-2493S.