Adnan Menderes Üniversitesi Sağlık Bilimleri Fakültesi Dergisi

2587-2427

Aydin Adnan Menderes University

10.46237/amusbfd.1287877

Health Care Administration

Sağlık Kurumları Yönetimi

Investigation Of The Anti-Proliferative and Anti-Apoptotic Effects Of Boric Acid On Human Non-Small Cell Lung Cancer Cells Through The TGF-β Signaling Pathway

Borik Asitin Anti-Proliferatif ve Anti-Apoptotik Etkilerinin İnsan Küçük Hücreli Dışı Akciğer Kanseri Hücrelerinde TGF-β Sinyal Yolağı Üzerinden İncelenmesi

https://orcid.org/0000-0003-4856-2304

Semerci Sevimli

Tuğba

ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ

https://orcid.org/0000-0001-5516-027X

Ghorbani

Aynaz

ESKİŞEHİR OSMANGAZİ ÜNİVERSİTESİ

https://orcid.org/0000-0001-8463-6943

Sevimli

Murat

SÜLEYMAN DEMİREL ÜNİVERSİTESİ, TIP FAKÜLTESİ

09 30 2023

7 3 553 564 04 26 2023 07 27 2023

2017

Adnan Menderes Üniversitesi Sağlık Bilimleri Fakültesi Dergisi

Objective: Lung cancer is the most common type of cancer in both men and women worldwide. Despite scientific advances in cancer in recent years, this pathology still ranks first with high mortality rates. Non-small cell lung cancer constitutes 85% of lung cancer cases. In these patients, the course of the disease progresses very poorly and since the diagnosis is mostly made in advanced stages, the treatments applied in most patients are not successful. Boric acid (BA) is an essential microelement with its chemical properties, low side effects and anti-carcinogenic effects. In this study, it was aimed to investigate the cytotoxic, anti-proliferative and apoptotic effects of Boric Acid (BA), which has known anti-proliferative and anti-apoptotic effects, on non-small cell lung cancer cells (A549) via TGF-β signaling pathway.Methods: Human non-small cell lung cancer cells (A549) were used in the study. Cytotoxic analysis of BA was performed by MTT analysis. The apoptotic effect of BA was determined by Annexin V/PI and immunofluorescence analysis. Expression levels of TGF-β, SMAD2/3/4 genes were analyzed at the molecular level by quantitative real-time polymerase chain reaction (RT-qPCR). Each reaction was replicated three times.Results: As a result of MTT analysis, it was observed that 50 mM BA decreased proliferation in A549 cells (p<0.01). With 50 mM BA application, the rate of apoptotic cells at 48 hours increased to 9.7%, and the rate of cells in the early apoptosis stage to 10.1%. In the immunofluorescence analysis, it was observed that the level of Caspase-3 labeling index in A549 cells was higher than the control group. No significant change was observed in the expression of SMAD3 and SMAD4 genes in BA applied A549 cells. While a decrease was observed in the expression of TGF-β gene (p<0.05), an increase in SMAD2 gene expression was observed (p<0.01).Conclusion: The results show that BA may be a new therapeutic agent with anti-proliferative and anti-apoptotic activity. However, further in vivo and advanced molecular level studies are needed to support these findings.

Amaç: Akciğer kanseri, tüm dünyada hem kadın hem de erkeklerde en sık görülen kanser türüdür. Son yıllarda kanser konusunda elde edilen bilimsel gelişmelere rağmen, bu patoloji hala yüksek mortalite oranları ile ilk sıralarda yer almaktadır. Akciğer kanseri olgularının %85’ini küçük hücreli olmayan akciğer kanseri oluşturmaktadır. Bu hastalarda hastalığın seyri oldukça kötü ilerlemekte ve tanısı çoğunlukla ileri evrelerde konulduğundan, çoğu hastada uygulanan tedaviler başarılı olamamaktadır. Borik asit (BA) sahip olduğu kimyasal özellikler, düşük yan etki oranları ve anti-kanserojen etkilere sahip esansiyel bir mikro elementtir. Bu çalışma kapsamında anti-proliferatif ve anti-apoptotik etkileri bilinen BA’nın, küçük hücreli dışı akciğer kanseri hücreleri (A549) üzerindeki sitotoksik, anti-proliferatif ve apoptotik etkilerinin TGF-β sinyal yolağı üzerinden araştırılması amaçlanmıştır.Yöntem: Çalışmada insan küçük hücreli dışı akciğer kanseri hücreleri (A549) kullanıldı. BA’nın sitotoksik analizi MTT analizi ile yapıldı. BA’nın apoptotik etkisi Annexin V/PI ve immünfloresan analizlerle belirlendi. TGF-β, SMAD2/3/4 genlerinin ekspresyon düzeyleri moleküler düzeyde kantitatif gerçek zamanlı polimeraz zincir reaksiyonu (RT-qPCR) ile analiz edildi. Her bir reaksiyon üç kez tekrarlandı.Bulgular: MTT analizi sonucu 50 mM BA’nın A549 hücrelerinde proliferasyonu azalttığı gözlendi (p<0.01). 50 mM BA uygulaması ile 48. saatte apoptotik hücre oranı %9.7’ye, erken apoptoz evresindeki hücre oranı ise %10.1’e yükseldi. İmmünfloresan analizinde de A549 hücrelerinde Kaspaz-3 işaretlenme düzeyininin kontrol grubuna göre yüksek miktarda olduğu gözlendi. BA uygulanan A549 hücrelerinde SMAD3 ve SMAD4 genlerinin ifadesinde anlamlı bir değişme gözlenmedi. TGF-β geninin ifadesinde azalma gözlenirken (p<0.05), SMAD2 gen ifadesinde artış gözlendi (p<0.01). Sonuç: Sonuçlar BA’nın; anti-proliferatif ve anti-apoptotik aktiviteye sahip yeni bir tedavi edici ajan olabileceğini göstermektedir. Ancak bu etkilere aracılık eden yolakların aydınlatılması için ileri düzeyde çalışmalara ihtiyaç bulunmaktadır.

Küçük hücreli dışı akciğer kanseri borik asit sitotoksisite apoptoz TGF-β/Smad sinyal yolağı

Non-small cell lung cancer boric acid cytotoxicity apoptosis TGF-β/Smad signaling pathway

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