Boric Acid-Induced Apoptosis in Breast Cancer Subtypes: Comparative Gene Expression and Machine Learning Study

Abstract

Aim: Breast cancer is a diverse illness that arises from the interplay of outside factors and genetic predisposition. Recently, artificial intelligence approaches have enabled the systematic analysis of potential anticancer agents such as boric acid by modeling the functions of apoptotic regulators. This study proposes an artificial intelligence-based analytical approach to evaluate how boric acid (B) affects cell survival and cell death through apoptosis pathway modulation.

Method: Viability of cells was determined by applying boric acid at various concentrations of 0.1, 1, 10, 50, 100, and 1000 ng/mL to cells at 24 h and 72 h using the cell viability test. Lastly, total RNA was isolated at 24 and 72 hours, and messenger RNA (mRNA) expression levels of specific genes were established by quantitative PCR (qPCR). An analysis was carried out using artificial intelligence based upon the Extreme Gradient Boosting (XGBoost) solution, a technique that enables the development of a pair of predictive models for cell viability and relative mRNA expression.

Results: After 24- and 72-hour treatments, boric acid reduced cell viability in MCF-7 and MDA-MB-231 cells in a rate- and time-sensitive manner in relation to the unaffected category. The triple-negative breast cancer phenotype was shown to be more susceptible to apoptosis at both time periods, as evidenced by the increased lethal consequences of boric acid in MDA-MB-231 cells compared to MCF-7 cells. However, boric acid was found to induce both intrinsic and extrinsic apoptosis in both breast cancer cell lines. Specifically for internal apoptosis signaling, ENDOG, BAX, Caspase-9, and Caspase-3 mRNA expression increased, while BCL-2 mRNA expression decreased, and the BAX/BCL-2 ratio was determined to shift towards apoptosis. Furthermore, apoptotic effects were detected in MCF-7 cells at 24 and 72 hours into the experiment, with this effect being particularly higher in MDA-MB-231 cells at 24 hours. The results obtained showed that boric acid induced the extrinsic apoptosis pathway by increasing mRNA FAS, FASLG, and CASP8 mRNA expression levels at 72 hours in MCF-7 cells and at 24 hours in MDA-MB-231 cells. However, boric acid increased MYC expression in MDA-MB-231 cells at 24 hours, while this effect was observed in MCF-7 cells at 72 hours of the experiment. The optimized XGBoost models were insufficient and showed highly non-homogeneous Margin of Deviation (MoD) values, with cell viability in the range of ±100% and mRNA expression spanning -4000% to +2000%.

Conclusions: The results obtained indicate that boric acid regulates cellular survival and death in breast cancer subtypes through MYC-dependent mechanisms and that this effect exhibits a more pronounced antitumor effect in MDA-MB-231 cells. So, it is concluded that boric acid may be a promising therapeutic agent for breast cancer if supported by additional research. Moreover, the inability of the current AI models also accentuates the complexity and non-linearity of the cellular response and suggests that the quantitative biological outcome can be better predicted with a broader multi-feature set.

Keywords

• Boric acid
• MCF-7
• MDA-MB-231
• XGBoost

Meme Kanseri Alt Tiplerinde Borik Asit ile İndüklenen Apoptoz: Karşılaştırmalı Gen Ekspresyonu ve Makine Öğrenmesi Çalışması

Abstract

Amaç: Meme kanseri, dış faktörlerin ve genetik yatkınlığın etkileşimi sonucu ortaya çıkan çok yönlü bir hastalıktır. Son zamanlarda yapay zeka yaklaşımları, apoptotik düzenleyicilerin işlevlerini modelleyerek borik asit gibi potansiyel antikanser ajanlarının sistematik analizini mümkün kılmaktadır. Bu çalışma, borik asit (B)’in apoptoz yolak modülasyonu üzerinden hücre sağkalımı ve hücre ölümünü nasıl etkilediğini değerlendirmek amacıyla yapay zeka tabanlı analitik bir yaklaşım önermektedir.

Yöntem: Hücre canlılığı, MCF-7 ve MDA-MB-231 meme kanseri hücre hatlarında 0.1, 1, 10, 50, 100 ve 1000 ng/mL konsantrasyonlarında borik asit uygulamasından 24 ve 72 saat sonra hücre canlılığı testi ile değerlendirildi. Son olarak, 24 ve 72 saatte toplam RNA izole edildi ve belirli genlerin mRNA ekspresyon seviyeleri kantitatif PCR (qPCR) ile belirlendi. Deneysel kantifikasyon için yapay zeka tabanlı bir analitik yaklaşım kullanıldı. Sonrasında hücre canlılığı ve mRNA ekspresyonunun iki bağımsız tahmin modelini oluşturmak için Extreme Gradient Boosting (XGBoost) çerçevesi kullanıldı.

Bulgular: Deneyin 24 ve 72. saatlerinde, borik asit, kontrol grubuna kıyasla, doz ve zamana bağlı olarak MCF-7 ve MDA-MB-231 hücre canlılığında azalmaya yol açtı. Borik asidin sitotoksik etkileri, MDA-MB-231 hücrelerinde MCF-7 hücrelerine göre daha güçlü gözlemlendi, bu da her iki zaman noktasında da üçlü negatif meme kanseri fenotipinde apoptoza karşı daha yüksek bir duyarlılığa işaret etmektedir. Bununla birlikte borik asit, her iki meme kanseri hücresinin iç ve dış apoptozunu indüklediği tespit edilmiştir. Özellikle iç apoptoz sinyallemesi için, ENDOG, BAX, Caspase-9 ve Caspase-3 mRNA ekspresyonu artarken, BCL-2 mRNA ekspresyonu ise azaldı ve BAX/BCL-2 oranının apoptoza doğru yön değiştirdiği belirlendi. Ayrıca apoptotik etkiler, deneyin 24. ve 72. Saatlerinde MCF-7 hücrelerinde belirlenirken özellikle bu etkinin deneyin 24. saatinde MDA-MB-231 hücrelerinde daha yüksek olduğu ortaya çıktı. Elde edilen sonuçlar, borik asitin MCF-7 hücrelerinde 72. saatte ve MDA-MB-231 hücrelerinde ise 24. saatte mRNA FAS, FASLG ve CASP8 mRNA ekspresyon seviyelerinin artarak dış apoptoz yolunu indüklediğini gösterdi. Bununla birlikte borik asit, MDA-MB-231 hucrelerinde 24. saatte MYC ekspresyonunu artırırken bu etki MCF-7 hücrelerinde deneyin 72. saatinde belirlendi. Çalışma, optimize edilmiş XGBoost modellerinin tahmin doğruluğunun yetersiz olduğunu, hem hücre canlılığı (±%100'e kadar) hem de mRNA ifadesi (-%4000'den +%2000'e kadar değişen) için son derece tutarsız sapma marjı değerlerine sahip olduğunu ortaya koydu.

Sonuç: Elde edilen sonuçlar, borik asidin meme kanseri alt tiplerinde hücresel sağkalım ve ölümü MYC-bağımlı mekanizmalar üzerinden düzenlediğini ve bu etkinin MDA-MB-231 hücrelerinde daha belirgin antitümoral etki ortaya koyduğunu göstermektedir. Bu nedenle, daha ileri gelişmiş deneylerle desteklenen borik asit, meme kanserini tedavi etmek için kullanılabilecek potansiyel bir terapötik ajandır. Ayrıca, mevcut yapay zeka modellerinin başarısızlığı, hücresel yanıtın son derece karmaşık ve son derece doğrusal olmayan bir yapıya sahip olduğunu göstermekte, bu da nicel biyolojik sonuç tahminlerini mümkün kılan daha geniş, çok özellikli bir setin önemli bir gereklilik olduğunu ortaya koymaktadır.

Keywords

• Borik asit
• MCF-7
• MDA-MB-231
• XGBoost

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