Dukel Muzaffer, Fiskin Kayahan
Molecular Biology and Genetics Department, Faculty of Art and Science, Mehmet Akif Ersoy University, Burdur, Turkey.
Biology Department, Faculty of Science, Akdeniz University, Antalya, Turkey.
Int J Radiat Biol. 2023;99(2):340-354. doi: 10.1080/09553002.2022.2110326. Epub 2022 Aug 15.
PAKs proteins are speculated as new promising targets for cancer therapy due to their central role in many oncogenic pathways. Because PAKs proteins are very significant during carcinogenesis, we aimed to investigate the hypothesis that inhibition of PAKs with IPA-3 and PF-3758309 treatment could synergistically reduce colon carcinoma cell growth.
The cytotoxic effects of both drugs were determined by a cell viability assay. Cell cycle and apoptosis were analyzed by flow cytometry. The effects of inhibitor drugs on marker genes of apoptosis, autophagy, cell cycle, and DNA damage were tested via immunoblotting.
We found out the synergistic effect of these drugs in pair on five colon cancer cell lines. Combined treatment with IPA-3+PF-3758309 in SW620 and Colo 205 cells markedly suppressed colon formation and induced apoptosis, cell cycle arrest, and autophagy compared with treatment with each drug alone. Additionally, this combination sensitized colon cancer cells to ionizing radiation that resulted in inhibition of cell growth.
Collectively, our findings show for the first time that cotreatment of IPA-3 with PF-3758309 exhibits superior inhibitory effects on colon carcinoma cell growth via inducing DNA damage-related cell death and also enforces a cell cycle arrest.
PAKs蛋白在许多致癌途径中起核心作用，因此被推测为癌症治疗中有前景的新靶点。由于PAKs蛋白在致癌过程中非常重要，我们旨在研究用IPA-3和PF-3758309处理抑制PAKs是否能协同降低结肠癌细胞的生长。
通过细胞活力测定法确定两种药物的细胞毒性作用。采用流式细胞术分析细胞周期和细胞凋亡情况。通过免疫印迹法检测抑制剂药物对凋亡、自噬、细胞周期和DNA损伤标志物基因的影响。
我们发现这两种药物对五种结肠癌细胞系具有协同作用。与单独使用每种药物相比，在SW620和Colo 205细胞中联合使用IPA-3 + PF-3758309显著抑制结肠形成并诱导细胞凋亡、细胞周期停滞和自噬。此外，这种联合用药使结肠癌细胞对电离辐射敏感，从而抑制细胞生长。
总体而言，我们的研究结果首次表明，IPA-3与PF-3758309联合处理通过诱导DNA损伤相关的细胞死亡对结肠癌细胞生长具有卓越的抑制作用，并且还能导致细胞周期停滞。
