Comparison of gene expressions in individuals with G6PD Mediterranean mutation and hemolytic anemia

Başak Günaştı; Abdullah Tuli

Makale detayları

Comparison of gene expressions in individuals with G6PD Mediterranean mutation and hemolytic anemia

Yazarlar:

Başak Günaştı | Abdullah Tuli

Yayıncı:

Cukurova University

Ana Başlıklar:

Klinik Tıp Bilimleri

Dergi:

Cukurova Medical Journal Vol.48 Issue 2, pp.351-360

Belge Tipi:

Makale

Yayın Tarihi:

2/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001286347

pISSN:

2602-3032

Anahtar kelimeler:

Özet:

 Purpose: In our study, we aimed to clarify the relationship between Glucose-6-phosphate dehydrogenase G6PD enzyme kinetics and mRNA expression levels of the G6PD gene in Gd-Med patients with and without hemolytic anemia. Materials and Methods: The study group consisted of 30 cases with Gd-Med mutation and 30 cases with enzyme activity levels in the reference range. G6PD activity was determined by the Beutler method. G6PD enzyme was partially purified with DE-52 anion exchange resin, and its kinetic parameters were studied. Gd-Med mutation was genotyped by MboII enzyme digest and sequence analysis. The expression level of the G6PD gene was calculated according to the 2-ΔΔCt formula. Results: In our study, a significant difference was found between the KmNADP and KmG6P values of the cases with Gd-Med mutation and the control group. There was no significant difference between KmNADP and KmG6P values in Gd-Med mutated patients with and without hemolytic anemia. Gene expression results of 18 patients without hemolytic anemia were significantly higher than 12 patients with hemolytic anemia. In addition, there was a significant difference between these variables and the control group. Conclusion: It might be a possible explanation that the substrate binding site of the enzyme in cases with Gd-Med mutation may have undergone post-transcriptional or post-translational modifications, and therefore gene expression might be changed. As a further study, the decrease in gene expressions of patients with hemolytic anemia with Gd-Med mutation can be clarified by evaluating the promoter side of the gene.

 Purpose: In our study, we aimed to clarify the relationship between Glucose-6-phosphate dehydrogenase G6PD enzyme kinetics and mRNA expression levels of the G6PD gene in Gd-Med patients with and without hemolytic anemia. Materials and Methods: The study group consisted of 30 cases with Gd-Med mutation and 30 cases with enzyme activity levels in the reference range. G6PD activity was determined by the Beutler method. G6PD enzyme was partially purified with DE-52 anion exchange resin, and its kinetic parameters were studied. Gd-Med mutation was genotyped by MboII enzyme digest and sequence analysis. The expression level of the G6PD gene was calculated according to the 2-ΔΔCt formula. Results: In our study, a significant difference was found between the KmNADP and KmG6P values of the cases with Gd-Med mutation and the control group. There was no significant difference between KmNADP and KmG6P values in Gd-Med mutated patients with and without hemolytic anemia. Gene expression results of 18 patients without hemolytic anemia were significantly higher than 12 patients with hemolytic anemia. In addition, there was a significant difference between these variables and the control group. Conclusion: It might be a possible explanation that the substrate binding site of the enzyme in cases with Gd-Med mutation may have undergone post-transcriptional or post-translational modifications, and therefore gene expression might be changed. As a further study, the decrease in gene expressions of patients with hemolytic anemia with Gd-Med mutation can be clarified by evaluating the promoter side of the gene.

 Purpose: In our study, we aimed to clarify the relationship between Glucose-6-phosphate dehydrogenase G6PD enzyme kinetics and mRNA expression levels of the G6PD gene in Gd-Med patients with and without hemolytic anemia. Materials and Methods: The study group consisted of 30 cases with Gd-Med mutation and 30 cases with enzyme activity levels in the reference range. G6PD activity was determined by the Beutler method. G6PD enzyme was partially purified with DE-52 anion exchange resin, and its kinetic parameters were studied. Gd-Med mutation was genotyped by MboII enzyme digest and sequence analysis. The expression level of the G6PD gene was calculated according to the 2-ΔΔCt formula. Results: In our study, a significant difference was found between the KmNADP and KmG6P values of the cases with Gd-Med mutation and the control group. There was no significant difference between KmNADP and KmG6P values in Gd-Med mutated patients with and without hemolytic anemia. Gene expression results of 18 patients without hemolytic anemia were significantly higher than 12 patients with hemolytic anemia. In addition, there was a significant difference between these variables and the control group. Conclusion: It might be a possible explanation that the substrate binding site of the enzyme in cases with Gd-Med mutation may have undergone post-transcriptional or post-translational modifications, and therefore gene expression might be changed. As a further study, the decrease in gene expressions of patients with hemolytic anemia with Gd-Med mutation can be clarified by evaluating the promoter side of the gene.

 Amaç: Çalışmamızda, Gd-Akdenizli bireylerde hemolitik anemisi olan ve olmayan olguların Glukoz-6-fosfat dehidrogenaz G6PD enzimi kinetiği ve G6PD geninin mRNA ekspresyon seviyeleri arasındaki ilişkinin irdelenerek bu duruma açıklık getirmeyi hedefledik. Gereç ve Yöntem: Çalışma grubu, Gd-Akdeniz mutasyonu bulunan 30 olgu ve enzim aktivite düzeyi referans aralığında olan 30 olgudan oluşmaktadır. G6PD aktivitesi Beutler yöntemi ile tayin edilmiştir. G6PD enzimi DE-52 anyon değiştirici reçine ile kısmi olarak saflaştırılarak kinetik özellikleri çalışılmıştır. MboII enzim kesimi ve sekans analizi ile Gd-Akdeniz mutasyonu genotiplendirilmiştir. G6PD geninin ekspresyon seviyesi 2-ΔΔCt formülüne göre hesaplanmıştır. Bulgular: Çalışmamızda Gd-Akdeniz mutasyonu bulunan olguların KmNADP ve KmG6P değerleri ile G6PD enzim aktivitesi referans değerlerde olan olgular arasında anlamlı düzeyde fark bulundu. Hemolitik anemisi olan ve olmayan Gd-Akdeniz mutasyonuna sahip olguların ise KmNADP ve KmG6P değerleri arasında anlamlı fark bulunamadı. Hemolitik anemisi olmayan 18 hastanın gen ekspresyon sonuçları, hemolitik anemisi olan 12 hastaya göre anlamlı derecede yüksek olduğu görüldü. Ayrıca enzim aktivite düzeyi referans aralığında olan olgularla da bu değişkenler kıyaslandığında yine anlamlı farklılık olduğu belirlendi Sonuç: Gd-Med mutasyonu olan olgularda enzimin substrat bağlanma bölgesinin post-transkripsiyonel veya post-translasyonel modifikasyonlara uğramış olması ve bu nedenle gen ekspresyonunun değişmesi olası bir açıklama olabilir. İleri çalışma olarak Gd-Akdeniz mutasyonuna sahip hemolitik anemisi bulunan olguların gen ekspresyonlarındaki azalmayı geninin promotör bölgesi değerlendirilerek açıklık getirilebilir.

Makale detayları

Comparison of gene expressions in individuals with G6PD Mediterranean mutation and hemolytic anemia

Yazarlar:

Başak Günaştı | Abdullah Tuli

Yayıncı:

Cukurova University

Ana Başlıklar:

Klinik Tıp Bilimleri

Dergi:

Cukurova Medical Journal Vol.48 Issue 2, pp.351-360

Belge Tipi:

Makale

Yayın Tarihi:

2/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001286347

pISSN:

2602-3032

Enzyme Kinetics

Gene Expression

Glucose-6-phosphate Dehydrogenase

Hemolytic Anemia.

Enzim Kinetiği

Gen Ekspresyonu

Glukoz-6-fosfat Dehidrogenaz

Hemolitik Anemi

Sayı

Makale detayları

Yazarlar: Başak Günaştı | Abdullah Tuli

Yayıncı: Cukurova University

Ana Başlıklar: Klinik Tıp Bilimleri

Dergi: Cukurova Medical Journal Vol.48 Issue 2, pp.351-360

Belge Tipi: Makale

Yayın Tarihi: 2/2023

Belge Dili: İngilizce

Document ID (Org No.): 2001286347

pISSN: 2602-3032

Sayı

Yazarlar:

Başak Günaştı | Abdullah Tuli

Yayıncı:

Cukurova University

Ana Başlıklar:

Klinik Tıp Bilimleri

Dergi:

Cukurova Medical Journal Vol.48 Issue 2, pp.351-360

Belge Tipi:

Makale

Yayın Tarihi:

2/2023

Belge Dili:

İngilizce

Document ID (Org No.):

2001286347

pISSN:

2602-3032