Gambogic Acid Improves Cisplatin Resistance of Bladder Cancer Cells through the Epithelial-Mesenchymal Transition Pathway Mediated by the miR-205-5p/ZEB1 Axis.

IF 1.1 4区医学 Q4 MEDICAL LABORATORY TECHNOLOGY

Annals of clinical and laboratory science Pub Date : 2024-05-01

Yuxian Mei, Jun Xu, Wenhua Li, Shasha Chen

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引用次数: 0

Abstract

Objective: Bladder cancer (BC) is primarily treated with cisplatin-based chemotherapy, but the development of cisplatin resistance often leads to BC recurrence. This study is focused on assessing the potential of gambogic acid (GA) in mitigating BC cells' cisplatin resistance, along with an analysis of the underlying mechanism involved.

Methods: Cisplatin was administered to human bladder transitional cell carcinoma cells (T24) at various concentration gradients to induce cisplatin-resistant (T24-DDP) cells. Several experimental groups were set: T24 group, T24-DDP group, T24-DDP+DDP group, T24-DDP+GA group, T24-DDP+DDP+GA group, T24-DDP+DDP+GA+miR-NC group, and T24-DDP+DDP+GA+miR-205-5p inhibitor group. The cell counting kit-8 (CCK-8) assay, Transwell migration assay, and scratch assay were respectively carried out for assessment of cell proliferation, invasion, and migration. Western blot analysis was conducted for detection of the protein expression of E-cadherin, ZEB1, Vimentin, N-cadherin, LRP, MRP, and P-gp in the cells, while the relative expression level of miR-205-5p was determined by qRT-PCR.

Results: In comparison with the T24-DDP group, cells in the T24-DDP+GA group showed enhanced sensitivity to cisplatin. Furthermore, as indicated by CCK-8 assay, GA improved T24-DDP cells' sensitivity to cisplatin, potentiated the effects of cisplatin, and exerted an inhibitory effect on the invasion, proliferation, as well as migration of T24-DDP cells. Through Western blot analysis, GA was revealed to significantly inhibit the expression of N-cadherin, E-cadherin, and Vimentin, as well as that of cisplatin-resistant proteins MRP, P-gp, and LRP in BC cells. In addition, shown by further experiments, GA promoted miR-205-5p expression and simultaneously inhibited ZEB1 expression within the cells.

Conclusion: GA alleviates BC cells' cisplatin resistance through the epithelial-mesenchymal transition pathway mediated by the miR-205-5p/ZEB1 axis.

本刊更多论文

甘草酸通过 miR-205-5p/ZEB1 轴介导的上皮-间质转化途径改善膀胱癌细胞对顺铂的耐药性

目的：膀胱癌（BC）主要采用顺铂化疗，但顺铂耐药性的产生往往会导致BC复发。本研究的重点是评估甘草酸（GA）在减轻膀胱癌细胞顺铂耐药性方面的潜力，并分析其中的内在机制：方法：以不同浓度梯度向人类膀胱过渡细胞癌细胞（T24）施用顺铂，诱导顺铂耐药细胞（T24-DDP）。实验分为几组：T24组、T24-DDP组、T24-DDP+DDP组、T24-DDP+GA组、T24-DDP+DDP+GA组、T24-DDP+DDP+GA+miR-NC组和T24-DDP+DDP+GA+miR-205-5p抑制剂组。分别采用细胞计数试剂盒-8（CCK-8）检测法、Transwell 迁移检测法和划痕检测法评估细胞增殖、侵袭和迁移。通过 Western 印迹分析检测细胞中 E-cadherin、ZEB1、Vimentin、N-cadherin、LRP、MRP 和 P-gp 的蛋白表达，并通过 qRT-PCR 检测 miR-205-5p 的相对表达水平：结果：与 T24-DDP 组相比，T24-DDP+GA 组细胞对顺铂的敏感性增强。此外，CCK-8 检测表明，GA 提高了 T24-DDP 细胞对顺铂的敏感性，增强了顺铂的作用，并对 T24-DDP 细胞的侵袭、增殖和迁移有抑制作用。通过 Western 印迹分析，GA 能显著抑制 BC 细胞中 N-cadherin、E-cadherin 和 Vimentin 的表达，以及顺铂抗性蛋白 MRP、P-gp 和 LRP 的表达。此外，进一步的实验表明，GA能促进miR-205-5p的表达，同时抑制细胞内ZEB1的表达：结论：GA可通过miR-205-5p/ZEB1轴介导的上皮-间质转化途径缓解BC细胞的顺铂耐药性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Annals of clinical and laboratory science 医学-医学实验技术

CiteScore

1.60

自引率

0.00%

发文量

112

审稿时长

6-12 weeks

期刊介绍： The Annals of Clinical & Laboratory Science welcomes manuscripts that report research in clinical science, including pathology, clinical chemistry, biotechnology, molecular biology, cytogenetics, microbiology, immunology, hematology, transfusion medicine, organ and tissue transplantation, therapeutics, toxicology, and clinical informatics.