The multifaceted antitumor effects of the ALK5 inhibitor BI 4659 in colon cancer: an in vitro study of cytotoxicity, clonogenicity, migration, and 3D spheroid formation

Abstract

Purpose: Inhibition of ALK5, the main receptor of the TGF-β pathway, is one of the new strategies in colon cancer treatment. In this study, the anticancer effects of the small molecule BI 4659 were investigated in the colon cancer cell line Caco-2. Method: Caco-2 cells were treated with increasing doses of BI 4659 for 72 hours. WST-1 was used for the viability assay, colony formation was used to observe the long-term effect, wound healing was used to determine migration ability, and hanging drop assays were performed for 3D tumors. Findings: BI 4659 exhibited high cytotoxicity in a dose-dependent manner in the Caco-2 cell line (IC50 = 29.33 ± 2.35 nM). Treatment resulted in significantly reduced colony formation, migration ability, and 3D spheroid size (p <0.05). Conclusion: Treatment with BI 4659 produced a multifaceted antitumor effect in the Caco-2 cell line. Inhibition of ALK5 showed an inhibitory effect on proliferation, migration, and 3D organization. These results suggest that BI 4659 may be a potential candidate for colon cancer treatment. Further experiments are recommended to elucidate the underlying molecular mechanisms and demonstrate vivo effects.

Keywords

• colon cancer
• ALK5 inhibitor
• BI 4659
• TGF-β
• cell migration
• tumor sprehoids

Kolon kanserinde ALK5 inhibitörü BI 4659'un çok yönlü antitümör etkileri: sitotoksisite, klonojeniklik, göç ve 3D sferoid oluşumu üzerine bir in vitro çalışma

Abstract

Amaç: TGF- β yolunun ana reseptörü olan ALK5’in inhibe edilmesi, kolon kanseri tedavsinde yeni stratejilerden birisidir. Bu çalışmada kolon kanser hücre hattı Caco-2’de küçük moleküllü BI 4659’un antikanser etkileri incelendi. Metod: Caco-2 hücreleri artan dozlarda BI 4659 ile 72 saat süre ile tedavi edildi. Canlılık deneyi için WST-1, uzun süreli etkiyi görmek için koloni formasyon, migrasyon yeteneğini belirlemek için Wound healing, 3B tümör için ise hanging drop deneyleri yapıldı. Bulgular: BI 4659, Caco-2 hücre hattında doza bağlı olarak yüksek sitotoksisite gösterdi (IC50 = 29.33 ± 2.35 nM). Tedavi ile koloni oluşumu, migrasyon yeteneği ve 3B sferoit büyüklüğü anlamlı düzeyde düşük çıktı (p <0.05). Sonuç: BI 4659 tedavisiyle Caco-2 hücre hattında çok yönlü antitümör etki ortaya çıkmıştır. ALK5’in inhibisyonu proliferasyon, göç ve 3B organizasyonunda baskılayıcı etki göstermiştir. Bu sonuçlar kolon kanseri tedavisinde BI 4659’un potansiyel bir aday olabileceğini göstermektedir. Altta yatan moleküler mekanizmaların ve in vivo etkilerin gösterilmesi için ileri deneylerin yapılması tavsiye edilmektedir.

Keywords

• kolon kanseri
• ALK5 inhibitörü
• BI 4659
• TGF-β
• hücre göçü
• tümör sferoidleri

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