Işıl TEKMEN, Doğan ÖZYURT, Çetin PEKÇETİN, ŞERMİN GENÇ, Kemal Kürşad GENÇ, Zişan BULDAN

TRAIL molekülünün in vitro beta hücre kültüründe hücre canlılığı üzerine etkisi

Amac,: Insüline bagimlı (Tip 1) diabetes mellitus (T1DM) pankreas beta adacik otoantijenlerine spesifik otoreaktif T hücrelerinin aracilı ettigi organa spesifik bir otoimmün hastaliktir. Bu hastalikta beta hücrelerinin %90'dan fazlasi kayba uğramaktadir ve bu hücre olümüne soluble ya da membrana bagk formda hücre olüm ligandlarınin aracilık etmesi olasidir. Bu molekiillerden biri tümor nekrozis faktor-alfa (TNFa) süperailesine ait bir hücre olüm ligandi olan TNF-related apoptosis inducing ligand (TRAIL) olabilir. Bu çalışmada TRAIL molekülünün erişkin siçan pankreas beta hücre kültürlerinde ve INS-IE sifan insulinoma hücre hatti kültürlerinde sitotoksik etkiye yol acip açmadiğının değerlendirilmesi amaçlanmıştir. Gereç ve yöntemler: Bu çalişmada hücre hasan modeli 10, 100 ve 1000 ng kon-santrasyonlarda TRAIL ile olusturuldu. Hücre canliliginın degerlendirilmesinde 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) testi kullanildi. Bulgular: Herhangi bir toksik madde eklenmeyen kontrol kültürleriyle karsilastırıldiginda TRAIL eklenen kültürlerde hücre canliliginın istatistiksel olarak anlamlı fark gostermedigi saptandi. Sonug: Bu sonuçlar TRAIL molekülünün pankreas beta hücre kültürlerinde ve INS-IE sican insulinoma hiicre hatti kiiltiirlerinde anlamh sitotoksik etkiye yol acmadigini gostermektedir. Bu hiicre kultiirii sisteminde TRAIL ile indiiklenen hücre olümüne direncin mekanizmalarının arastırılması açisindan INS-IE hiicre hatti hücrelerinde TRAIL reseptorlerinin ve dogal apoptoz inhibitorü proteinlerin ekspresyonunun belirlenmesi uygun olacaktir.

Anahtar Kelimeler:

Hücre sağkalımı, Diabetes mellitus, tip 1, Tümör nekrozis faktör-alfa, İnsülin salgılayan hücreler, Apoptozis düzenleyici proteinler

The effect of TRAIL molecule on cell viability in in vitro beta cell culture

Objective: Insulin-dependent (Type 1) diabetes mellitus (T1DM) is an organ-specific autoimmune disease which is mediated by autoreactive T cells that are specific against pancreatic beta islet autoantigens. In this disorder, above 90% of beta cells undergo loss and it is likely that soluble or membrane-bound cell death ligands mediate beta cell loss. One of the these putative molecules may be TNF-related apoptosis inducing.ligand (TRAIL) that is a cell death ligand and member of TNF superfamily. In this study, it was aimed to evaluate whether TRAIL molecule results in cytotoxicity in adult rat pancreatic beta cell cultures and INS-IE rat insulinoma cell line cultures. Material and methods: In the present study, cell injury was induced by TRAIL at 10, 100 and 1000 ng concentrations. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetra- zolium bromide (MTT) test was used to evaluate cell viability. Results: In the present study, it was found that TRAIL does not induce statistically significant cytotoxicity in beta cell and INS-IE cell line cultures. Discussion: The results of the present study show that TRAIL does not result significant cytotoxicity in pancreas beta cell and INS-IE rat insulinoma cell line cultures. To clarify the mechanisms of resistance of this cell line against TRAIL- induced cell death, the expression of TRAIL receptors and natural apoptosis inhibitpr proteins should be evaluated in beta cell cultures and INS-IE cell line.

Keywords:

Cell Survival, Diabetes Mellitus, Type 1, Tumor Necrosis Factor-alpha, Insulin-Secreting Cells, Apoptosis Regulatory Proteins,

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