Metin AÇIKYILDIZ, Erdal SÖNMEZ, Abdulgani TATAR, Mehmet Enes ARSLAN, Hasan TÜRKEZ, Fatime GEYİKOĞLU

Mikrodizin Analizi ile Demir Oksit Nanoparçaciklarinin İnsan Akciğer Epitel Hücre Hatti Üzerinde Ekspresyon Profil ve Yolak Analizleri

Toksikogenomik, kanser, immünolojik hastalıklar, çevresel tepkiler, gen-gen etkileşimleri ve ilaç toksisitesi gibi hücresel yolları ve mekanizmaları araştıran gelişmekte olan bir alandır. Nanopartiküller (NP’ler), çeşitli biyolojik aktivitelerde sıradışı özelliklerinden dolayı toksikogenomik deneyler için önemli hale gelmişlerdir. Bu nedenle yapılan çalışmada insan alveolar epitel hücrelerinin (HPAEpiC) gen ekspresyonu profillemesi üzerinde demir oksidin (Fe2O3) nanotoksisitesi incelenmiştir. Bu amaçla, demir oksit nanopartikülleri kısmi erime metodu ile sentezlenmiş ve X-ışını kristalografisi (XRD) ve transmisyon elektron mikroskobu (TEM) teknikleri kullanılarak karakterize edilmiştir. Hücre canlılığı ve sitotoksisite 3- (4,5-dimetil-tiazol-2-il) 2,5-difeniltetrazolyum bromür (MTT), nötral kırmızı (NR) ve laktat dehidrojenaz (LDH) salım testleri ile tespit edilmiştir. Bütün genom mikrodizin analizi, demir oksidin nanopartikülleriyle kültürlenmiş insan alveolar epitel hücrelerinde gen ekspresyonu üzerindeki etkilerini araştırmak için uygulanmıştır. Bunların yanısıra, mikrodizin sonucunda elde edilen farklı ekspresyon düzeylerine sahip genler, gen ontolojisi (GO) analizi ile DAVID (Annotation, Visualization ve Integrated Discovery Veri Tabanı) kullanılarak işlevsel olarak sınıflandırılmıştır. Bu çalışmadan elde edilen sonuçlar, demir oksidin doğrudan veya dolaylı olarak hücre proliferasyonunun düzenlenmesini, hormon uyarımına tepkiyi, östrojen uyarısını, sitokin aktivitesini ve çeşitli genleri uyararak kan dolaşımını etkilediğini göstermiştir.

Expression Profiling and Pathway Analysis of Iron Oxide Nanoparticles Toxicity on Human Lung Alveolar Epithelial Cell Line Using Microarray Analysis

Toxicogenomics is a developing area searching for cellular pathways and mechanisms including cancer, immunological diseases, environmental responses, gene-gene interactions and drug toxicity. Nanoparticles (NPs) become important candidates for analyzing in toxicogenomic experiments because of their unusual properties in various biological activities. Therefore, we examined the nanotoxicity of iron oxide (Fe2O3) on gene expression profiling of human alveolar epithelial cells (HPAEpiC) in the study. For this aim, iron oxide nanoparticles were synthesized by zone melting method and characterized via using X-ray crystallography (XRD) and transmission electron microscope (TEM) techniques. Cell viability and cytotoxicity were determined by 3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT), neutral red (NR) and lactate dehydrogenase (LDH) release tests. Whole-genome microarray expression analysis was performed to explore the effects of iron oxide nanoparticles on gene expression in cultured human alveolar epithelial cells. For further analyses, these genes were functionally classified by using DAVID (The Database for Annotation, Visualization and Integrated Discovery) with gene ontology (GO) analysis. The results from this study indicated that iron oxide-mediated toxicity directly or indirectly affecting the regulation of cell proliferation, response to hormone stimulus, estrogen stimulus, cytokine activity and blood circulation by stimulating diverse genes.

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