Metformin-induced and Mitochondrial Stress-mediated Apoptosis in Schizosaccharomyces pombe

Year 2024, Volume: 11 Issue: 1, 174 - 182, 31.05.2024

Hızlan Hıncal Ağuş Cenk Kığ Mustafa Kaçmaz

https://doi.org/10.35193/bseufbd.1329191

Abstract

Metformin, a widely used first-line medication in the treatment of type II diabetes, has been proposed to have a second indication in the treatment of cancers and aging. However, its accounting mechanisms in cellular physiology were not clearly understood. Therefore, its cytotoxicity and underlying physiological mechanisms should be explained. Schizosaccharomyces pombe was evaluated as a single-cell cytotoxicity model and was treated with metformin and grown on YEL media at 30 °C and 180 rpm. 0,1-20 mM metformin caused dose-dependent apoptosis and necrosis demonstrated by using Annexin V-FITC/PI and DAPI staining. Surprisingly, metformin reduced ROS levels with stable antioxidant enzyme levels, but the mitochondrial transmembrane potential was significantly increased indicating a differential regulation by the dual character of metformin. In addition, a possible role can be attributed to Cnx1 in apoptotic cell death; which showed a dramatic increase in transcription, however, three other potential apoptotic genes, Rad9, Pca1, and Aif1 were stable. To conclude, the dual effect of metformin was clarified, and related cellular physiological effects with accompanying mechanisms (particularly Cnx1-mediated) were shown using S. pombe.

Keywords

S. pombe, Metformin, Mitochondrial Stress, Apoptosis

Project Number

119Z186

Schizosaccharomyces pombe’de Metformin-tetiklemeli ve Mitokondriyal Stres Aracılı Apoptoz

Abstract

Tip II diyabetin tedavisinde ilk müdahale olarak yaygın kullanılan metforminin kanser tedavisi ve yaşlanma için sekonder endikasyona sahip olduğu öne sürülmüştür. Ancak hücresel fizyolojik sorumlu mekanizmalar net olarak anlaşılmamıştır. Bu yüzden, sitotoksik ve fizyolojik mekanizmaların açıklığa kavuşturulması gerekmektedir. Schizosaccharomyces pombe tek hücreli sitotoksisite modeli olarak değerlendirilmiş, metforminle muamele edilerek 30 °C sıcaklık ve 180 rpm hızda, YEL medyumu içerisinde inkübasyona bırakılmıştır. Annexin V-FITC/PI ve DAPI boyaması ile gösterildiği üzere, 0,1-20 mM metformin doz-bağımlı apoptoz ve nekroza neden olmuştur. Öte yandan, metformin ROS seviyelerini düşürmüş, uygulamada antioksidan enzim seviyeleri sabit kalmış, fakat mitokondri membran potansiyeli önemli derecede yükselmiştir. Bu durum metforminin ikili karakteri ile diferansiyel regülasyona işaret etmektedir. Ek olarak, apoptotik hücre ölümünde Cnx1’e olası bir rol biçilebilir; ki transkripsiyonda dramatik bir yükseliş görülmüştür. Ancak diğer üç olası apoptoz geni Rad9, Pca1 ve Aif1 stabil kalmıştır. Sonuç olarak, bu çalışma ile metforminin ikili etkisi net olarak ortaya konmuş ve sorumlu mekanizmalarla birlikte (özellikle Cnx1-aracılı), ilişkili hücresel fizyolojik etkiler S. pombe maya hücresinde gösterilmiştir.

Keywords

Schizosaccharomyces pombe, metformin, mitokondriyal stress, apoptoz

Supporting Institution

TÜBİTAK

Project Number

119Z186

Thanks

This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK) 1002 Program (119Z186). We especially thank B. PALABIYIK and E. YORUK for the chemicals and S. pombe strains.

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