Evaluation of the Anti-Cancer Effect of TRAF2 and NCK In teracting Protein Kinase (TNIK) Inhibition in Breast Cancer Cells

Abstract

Objective: This study aimed to evaluate the anticancer effect of NCB-0846, a TNIK inhibitor, in MCF-7 cells and to assess its impact on the expression levels of NF-κB and TNFA at the gene level. Materials and Methods: The MCF-7 cell line was cultured at 37°C in a 5% CO2 atmosphere using Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics (50 IU/mL penicillin and 50 mg/mL streptomycin). Cell viability was analyzed using the CCK-8 assay to determine the cytotoxic effect of NCB- 0846. Acridine Orange/Propidium Iodide (AO/PI) staining was performed to evaluate the effect of NCB-0846 on cellular morphology in the MCF-7 cell line. Total RNA was isolated from cells treated with NCB-0846. Data analysis was performed using the SPSS 22.0 statistical program. Results: The data indicated that NCB-0846 significantly decreased the viability rates of MCF-7 cells in a dose-dependent manner (1-3 μM, p<0.01). RT-PCR analysis revealed that the expression level of NFKB1 increased 5.4-fold compared to the control group in MCF-7 cells treated with NCB-0846 at the effective dose and duration (p<0.01). In contrast, the expression level of TNFA decreased to 0.4-fold compared to the control group (p<0.01). Conclusion: The results demonstrate that NCB-0846 induces changes in the mRNA levels of the NFKB1 and TNFA genes, which are associated with inflammatory signalling pathways in MCF-7 cells. However, further molecular analyses are necessary to clarify the effect of NCB-0846 on inflammation in breast cancer and other cancer types.

Keywords

• Breast cancer
• Inflammation
• NCB-0846
• TNIK

Meme Kanseri Hücrelerinde TRAF2 ve NCK Etkileşimli Protein Kinaz (TNIK) İnhibisyonunun Kanser Karşıtı Etkisinin Değerlendirilmesi

Abstract

Amaç: Bu çalışmanın amacı, TNIK inhibitörü olan NCB-0846'nın MCF-7 hücrelerindeki antikanser etkisini değerlendirmek ve gen düzeyinde NF-κB ve TNFA ifade seviyeleri üzerindeki etkisini değerlendirmektir. Materyal ve Metot: MCF-7 hücre hattı, %10 fetal sığır serumu (FBS) ve antibiyotiklerle (50 IU/mL penisilin ve 50 mg/mL streptomisin) desteklenmiş Dulbecco's Modified Eagle Medium (DMEM) kullanılarak %5 CO2 atmosferinde 37°C'de kültüre edildi. Hücre canlılığı, NCB-0846'nın sitotoksik etkisini belirlemek için CCK-8 testi kullanılarak analiz edildi. Akridin Oranj/Propidyum İyodür (AO/PI) boyama, NCB-0846'nın MCF-7 hücre hattındaki hücresel morfoloji üzerindeki etkisini değerlendirmek için gerçekleştirildi. Toplam RNA, NCB-0846 ile tedavi edilen hücrelerden izole edildi. Veri analizi SPSS 22.0 istatistik programı kullanılarak gerçekleştirildi. Bulgular: Veriler, NCB-0846'nın MCF-7 hücrelerinin canlılık oranlarını doza bağlı bir şekilde önemli ölçüde azalttığını gösterdi (1-3 µM, p<0,01). RT-PCR analizi, etkili doz ve sürede NCB-0846 ile tedavi edilen MCF-7 hücrelerinde NFKB1 ekspresyon düzeyinin kontrol grubuna kıyasla 5,4 kat arttığını ortaya koydu (p<0,01). Buna karşılık, TNFA ekspresyon düzeyi kontrol grubuna kıyasla 0,4 kat azaldı (p<0,01). Sonuç: Sonuçlar, NCB-0846'nın MCF-7 hücrelerinde inflamatuar sinyal yollarıyla ilişkili olan NFKB1 ve TNFA genlerinin mRNA düzeylerinde değişikliklere neden olduğunu göstermektedir. Ancak, NCB-0846'nın meme kanseri ve diğer kanser türlerindeki inflamasyon üzerindeki etkisini açıklığa kavuşturmak için daha fazla moleküler analiz gereklidir.

Keywords

• Meme kanseri
• Enflamasyon
• NCB-0846
• TNIK

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