ISSN: 1301-2193 E-ISSN: 1308-9846
  • Turkish Journal of
    Endocrinology and Metabolism

Association of Serum Resistin Level and Resistin (RETN) Gene (-420 C>G) Polymorphism in Pakistani Women with Polycystic Ovarian Syndrome
Polikistik Over Sendromlu Pakistanlı Kadınlarda Resistin (RETN) Gen (-420 C>G) Polimorfizmi ve Serum Resistin Düzeyi İlişkisi
Received Date : 05 May 2019
Accepted Date : 18 Dec 2019
Available Online : 04 Feb 2020
University of Health Sciences, Department of Chemical Pathology, Lahore, Pakistan *University of Health Sciences, Department of Cell Biology and Physiology, Lahore, Pakistan **Jouf University, Department of Clinical Laboratory Sciences, Sakaka, Aljouf, Kingdom of Saudi Arabia
Doi: 10.25179/tjem.2019-66784 - Makale Dili: EN
Turk J Endocrinol Metab 2020;24:16-22
ABSTRACT
Objective: The objective of the present study was to investigate the association of altered serum resistin levels to RETN gene (420C>G) polymorphism in women with polycystic ovarian syndrome (PCOS) and in healthy controls. Material and Methods: Eighty (40 PCOS cases and 40 healthy controls) individuals were included. Whole blood and serum samples were taken from all participants. Enzyme linked immunosorbent (ELISA) was performed for measuring the levels of serum resistin. Whole blood was used for extracting total genomic DNA by the phenol-chloroform method. Polymerase chain reaction with fragment length polymorphism was performed for detecting single nucleotide polymorphism (SNP) in the promoter region (-420 C>G) of the resistin (RETN) gene by amplifying the oligonucleotide sequence of the SNP. The amplified products were first confirmed on 2.0% agarose gel for product size, and then restriction digestion of these products was performed by using the Bpil restriction enzyme. After completion of digestion, the products were resolved on 2.5% agarose gel with a 100 bp DNA ladder, and the bands were inspected to infer genotype. Data analysis was done using SPSS software and the association between serum resistin levels and RETN genotypes was analyzed. Results: There was no significant difference (p=0.125) observed in serum resistin levels between PCOS cases (mean±SD=19.33±3.50) and healthy controls (mean±SD= 13.48±1.31). The frequency of the G allele was high in PCOS cases (65%) than in controls (53.7%). The GG genotype frequency of SNP (-420 C>G) was high in PCOS cases (40%) than in controls (20%), but no association was found (p=0.148). The high serum resistin levels were significantly associated with the GG genotype in PCOS cases (p=0.027). Conclusion: High serum resistin levels are not associated with the genotypes of RETN (-420 C>G) polymorphism in PCOS women and controls, although women with PCOS had high GG genotype levels of serum resistin. Further studies with large sample size should be conducted to explore the mechanism of genetic factors in complex diseases like PCOS.
ÖZET
Amaç: Polikistik over sendromu (PKOS) olan kadınlarda ve sağlıklı kontrollerde değişmiş serum resistin düzeylerinin RETN geni (–420C> G) polimorfizmi ile ilişkisini araştırmaktır. Gereç ve Yöntemler: Çalışmaya, seksen kişi (40 PKOS hastası ve 40 sağlıklı kontrol) dâhil edildi. Tüm katılımcılardan tam kan ve serum örnekleri alındı. Serum resistin düzeylerini ölçmek için enzime bağlı immünosorbent (ELISA) yapıldı. Tam kan, fenolkloroform yöntemiyle total genomik DNA’nın ekstrakte edilmesi için kullanıldı. Tek nükleotit polimorfizminin (SNP) oligonükleotid sekansını amplifiye ederek resistin (RETN) geninin promotör bölgesinde (-420 C>G) SNP saptanması için polimeraz zincir reaksiyonu ile parça uzunluk polimorfizmi yapıldı. Amplifiye edilen ürünler önce ürün boyutu için %2,0 agaroz jel üzerinde doğrulandı ve daha sonra bu ürünlerin sindirimi Bpil restriksiyon enzimi kullanılarak gerçekleştirildi. Sindirimin tamamlanmasından sonra ürünler, 100 bp'lik bir DNA merdiveni ile %2,5 agaroz jel üzerinde çözündürüldü ve bantlar genotip çıkarımı için incelendi. SPSS yazılımı kullanılarak veri analizi yapıldı ve serum resistin düzeyleri ile RETN genotipleri arasındaki ilişki analiz edildi. Bulgular: PKOS hastaları (ortalama±SS=19,33±3,50) ile sağlıklı kontroller (ortalama±SS=13,48±1,31) arasında serum resistin düzeylerinde anlamlı bir fark izlenmedi (p=0,125). G allelinin sıklığı PKOS (%65) hastalarında kontrollerden (%53,7) yüksekti. SNP’nin (-420 C>G) GG genotip frekansı, PKOS (%40) vakalarında kontrollerden (%20) yüksekti, ancak ilişki bulunmadı (p=0,148). Yüksek serum resistin düzeyleri PKOS hastalarında GG genotipi ile anlamlı şekilde ilişkili idi (p=0,027). Sonuç: PKOS’lu kadınların serum resistin GG genotip düzeyleri yüksek olsa da PKOS’lu kadınlarda ve kontrollerde yüksek serum resistin düzeyleri ile RETN (-420 C>G) polimorfizmi genotipleri ilişkili değildir. PKOS gibi karmaşık hastalıklarda genetik faktörlerin mekanizmasını araştırmak için büyük ölçekli daha ileri çalışmalar yapılmalıdır.
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