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

Yıl 2023, Cilt: 7 Sayı: 3, 553 - 564, 30.09.2023

Tuğba Semerci Sevimli Aynaz Ghorbani Murat Sevimli

https://doi.org/10.46237/amusbfd.1287877

Öz

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.

Anahtar Kelimeler

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

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

Öz

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.

Anahtar Kelimeler

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

Kaynakça

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