Sevim Nalkiran Hatice, Akcora Yildiz Dilara, Saydam Faruk, Guzel Ali Irfan, Nalkiran Ihsan
Department of Medical Biology, Faculty of Medicine, Recep Tayyip Erdogan University, Rize, Turkey.
Department of Biology, Faculty of Arts and Sciences, Mehmet Akif Ersoy University, Burdur, Turkey.
Saudi J Biol Sci. 2023 Mar;30(3):103564. doi: 10.1016/j.sjbs.2023.103564. Epub 2023 Jan 23.
Improving the chemotherapy sensitivity of bladder cancer is a current clinical challenge. It is critical to seek out effective combination therapies that include low doses of cisplatin due to its dose-limiting toxicity. This study aims to investigate the cytotoxic effects of the combination therapy including proTAME, a small molecule inhibitor, targeting Cdc-20 and to determine the expression levels of several APC/C pathway-related genes that may play a role in the chemotherapy response of RT-4 (bladder cancer) and ARPE-19 (normal epithelial) cells. The IC20 and IC50 values were determined by MTS assay. The expression levels of apoptosis-associated ( and and APC/C-associated , , and genes were assessed by qRT-PCR. Cell colonization ability and apoptosis were examined by clonogenic survival experiment and Annexin V/PI staining, respectively. Low-dose combination therapy showed a superior inhibition effect on RT-4 cells by increasing cell death and inhibiting colony formation. Triple-agent combination therapy further increased the percentage of late apoptotic and necrotic cells compared to the doublet-therapy with gemcitabine and cisplatin. ProTAME-containing combination therapies resulted in an elevation in Bax/Bcl-2 ratio in RT-4 cells, while a significant decrease was observed in proTAME-treated ARPE-19 cells. Cdc-20 expression in proTAME combined treatment groups were found to be decreased compared to their control groups. Low-dose triple-agent combination induced cytotoxicity and apoptosis in RT-4 cells effectively. It is essential to evaluate the role of APC/C pathway-associated potential biomarkers as therapeutic targets and define new combination therapy regimens to achieve improved tolerability in bladder cancer patients in the future.
提高膀胱癌的化疗敏感性是当前临床面临的一项挑战。由于顺铂具有剂量限制性毒性，因此寻找包括低剂量顺铂在内的有效联合疗法至关重要。本研究旨在探讨包括小分子抑制剂proTAME靶向Cdc-20的联合疗法的细胞毒性作用，并确定几种可能在RT-4（膀胱癌）和ARPE-19（正常上皮）细胞化疗反应中起作用的APC/C途径相关基因的表达水平。通过MTS法测定IC20和IC50值。通过qRT-PCR评估凋亡相关基因（ 、 和 ）以及APC/C相关基因（ 、 、 和 ）的表达水平。分别通过克隆形成存活实验和Annexin V/PI染色检测细胞集落形成能力和凋亡情况。低剂量联合疗法通过增加细胞死亡和抑制集落形成对RT-4细胞显示出更强的抑制作用。与吉西他滨和顺铂的双联疗法相比，三联疗法进一步增加了晚期凋亡和坏死细胞的百分比。含proTAME的联合疗法导致RT-4细胞中Bax/Bcl-2比值升高，而在proTAME处理的ARPE-19细胞中观察到显著降低。与对照组相比，proTAME联合治疗组中Cdc-20的表达降低。低剂量三联疗法有效地诱导了RT-4细胞的细胞毒性和凋亡。未来评估APC/C途径相关潜在生物标志物作为治疗靶点的作用并确定新的联合治疗方案以提高膀胱癌患者的耐受性至关重要。
