Cellular Therapy and Stem Cell Production Application and Research Centre, ESTEM, Eskisehir Osmangazi University, Eskisehir, Turkey.
Department of Stem Cell, Institute of Health Sciences, Eskisehir Osmangazi University, Eskisehir, Turkey.
Biotechnol J. 2024 Feb;19(2):e2300496. doi: 10.1002/biot.202300496.
Despite advances in early detection and treatment, colorectal cancer remains one of the leading causes of cancer-related deaths. The klotho (KL) gene plays a critical role in the development and progression of colorectal cancer. This study investigates the role of the KL gene in colorectal cancer by using the CRISPR/Cas9 system to overexpress and knock out (KO) the KL gene in human colorectal cancer cells (Caco-2). The effects of the changes were assessed by gene expression analysis, flow cytometry, scratch wound closure assays, colony formation assays, and immunofluorescence staining. Our results showed that overexpression of the KL gene increased apoptosis and decreased cell motility in cancer cells, whereas knockout of the KL gene had the opposite role. The present study elucidates the mechanisms underlying this role and highlights the potential of the CRISPR/Cas9 system as a gene editing tool in cancer research. Our data suggest that activation of the KL gene may serve as a novel therapeutic strategy and biomarker for studies in colorectal cancer.
尽管早期检测和治疗取得了进展，但结直肠癌仍然是癌症相关死亡的主要原因之一。 klotho（KL）基因在结直肠癌的发生和发展中起着关键作用。本研究通过使用 CRISPR/Cas9 系统过表达和敲除（KO）人结直肠癌细胞（Caco-2）中的 KL 基因，来研究 KL 基因在结直肠癌中的作用。通过基因表达分析、流式细胞术、划痕愈合试验、集落形成试验和免疫荧光染色来评估变化的影响。我们的结果表明，KL 基因的过表达增加了癌细胞的凋亡并降低了其运动性，而 KL 基因的敲除则产生了相反的作用。本研究阐明了这种作用的机制，并强调了 CRISPR/Cas9 系统作为癌症研究中基因编辑工具的潜力。我们的数据表明，KL 基因的激活可能成为结直肠癌研究中的一种新的治疗策略和生物标志物。
