Assessment of Anticancer Properties of Synthesized Pyrazole-Acridine Derivative on SH-SY5Y Human Neuroblastoma Cells

Year 2025, Issue: Early Access

Yusuf Küçükbağrıaçık , Muna Elmusa , Fatıma Elmusa , Hümeyra Yılmaz , Rahmi Kasımoğulları , Mohammadreza Dastourı

https://doi.org/10.18678/dtfd.1520993

Abstract

Aim: This study aimed to synthesize a novel pyrazole acridine derivative (3-ACH) and evaluate its anticancer properties on SH-SY5Y human neuroblastoma cells.
Material and Methods: The pyrazole-4-carbaldehyde derivative was cyclized with dimedone and p-nitroaniline to synthesize the 3-ACH. Characterization of the compound was performed using FT-IR, NMR, HPLC-Q-TOF/MS, and elemental analysis. The cytotoxic impact of the 3-ACH compound on SH-SY5Y human neuroblastoma cells was evaluated using the WST-1 assay in a dose- (50, 100, and 150 μg/mL) and time-dependent (12, 24, and 48 hours) manner. The impact of 3-ACH on apoptosis was investigated through immunostaining for Caspase-3, -8, and -9, and BAX proteins.
Results: 3-ACH reduced SH-SY5Y cell viability with the rate of 89.89±7.63% (p=0.002) at 100 μg/mL concentration after 24 hours of treatment. While a higher (150 μg/mL) concentration showed a similar reduction (90.53±2.88%, p=0.004), the lower (50 μg/mL) concentration maintained high cell viability (98.37±1.67%, p=0.903) at 24 hours. All doses of 50 μg/mL (94.55±0.65%), 100 μg/mL (95.18±1.41%), and 150 μg/mL (95.28±2.57%) significantly reduced cell viability rates, at 48 hours (all p<0.001). Immunostaining revealed a significant upregulation in the synthesis of BAX, Caspase-3, -8, and -9 proteins in cells treated with 3-ACH (100 μg/mL) for 24 hours compared to the control group.
Conclusion: These results indicate that 3-ACH has the potential to induce apoptosis in SH-SY5Y cells by activating both the intrinsic and extrinsic pathways, which may contribute to its cytotoxic effects. This study provides promising evidence supporting the potential of the 3-ACH compound as an anticancer therapeutic agent.

Keywords

Chemical synthesis, antineoplastic agents, neuroblastoma, pyrazoles, acridines, cytotoxicity, apoptosis

Sentezlenen Pirazol-Akridin Türevinin SH-SY5Y İnsan Nöroblastoma Hücrelerinde Antikanser Özelliklerinin Değerlendirilmesi

Abstract

Amaç: Bu çalışmanın amacı yeni bir pirazol akridin türevi (3-ACH) sentezlemek ve bu bileşiğin SH-SY5Y insan nöroblastoma hücreleri üzerindeki antikanser özelliklerini değerlendirmektir.
Gereç ve Yöntemler: Pirazol-4-karbaldehit türevi, 3-ACH sentezlemek için dimedon ve p-nitroanilin ile siklize edildi. Bileşiğin karakterizasyonu FT-IR, NMR, HPLC-Q-TOF/MS ve element analizi kullanılarak yapıldı. 3-ACH bileşiğinin SH-SY5Y insan nöroblastoma hücreleri üzerindeki sitotoksik etkisi, doz (50, 100 ve 150 μg/mL) ve zaman (12, 24 ve 48 saat) bağımlı bir şekilde WST-1 testi kullanılarak değerlendirildi. 3-ACH'nin apoptoz üzerindeki etkisi, Kaspaz-3, -8, -9 ve BAX proteinleri için immün boyama yolu ile araştırıldı.
Bulgular: 3-ACH, 100 μg/mL konsantrasyonda 24 saatlik tedaviden sonra SH-SY5Y hücre canlılığını %89,89±7,63 (p=0,002) oranında azaltmıştır. 24 saatte, daha yüksek (150 μg/mL) konsantrasyon benzer bir azalma gösterirken (%90,53±2,88, p=0,004), daha düşük (50 μg/mL) konsantrasyon hücre canlılığını korumuştur (%98,37±1,67, p=0,903). 50 μg/mL (%94,55±0,65), 100 μg/mL (%95,18±1,41) ve 150 μg/mL (%95,28±2,57) dozlarının tümü, 48 saatte hücre canlılık oranlarını önemli ölçüde azaltmıştır (tüm p<0,001). İmmün boyama, 24 saat boyunca 3-ACH (100 μg/mL) ile muamele edilen hücrelerde, BAX, Kaspaz-3, -8 ve -9 proteinlerinin sentezinde kontrol grubuna kıyasla önemli bir artış olduğunu ortaya koymuştur.
Sonuç: Bu sonuçlar, 3-ACH'nin hem intrinsik hem de ekstrinsik yolakları aktive ederek SH-SY5Y hücrelerinde, sitotoksik etkilerine katkıda bulunabilecek olan, apoptozu indükleme potansiyeline sahip olduğunu göstermektedir. Bu çalışma, 3-ACH bileşiğinin antikanser terapötik ajan olarak potansiyelini destekleyen umut verici kanıtlar sunmaktadır.

Keywords

Kimyasal sentez, antineoplastik ajanlar, nöroblastoma, pirazoller, akridinler, sitotoksisite, apoptoz

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