福美双作为 11β-羟甾脱氢酶 2 型抑制剂，通过 Wnt/β-连环蛋白通路增强了氢化可的松治疗骨肉瘤的抑制作用。

Thiram, an inhibitor of 11ß-hydroxysteroid dehydrogenase type 2, enhances the inhibitory effects of hydrocortisone in the treatment of osteosarcoma through Wnt/β-catenin pathway.

机构信息

Clinical Translational Innovation Center/Molecular Medicine Research Center, West China Hospital, Sichuan Univicity, Chengdu, Sichuan Province, 610041, People's Republic of China.

Division of of Radiotherapy, Cancer Center,West China Hospital, Sichuan University, Chengdu, Sichuan Province, 610041, People's Republic of China.

出版信息

BMC Pharmacol Toxicol. 2023 Mar 28;24(1):20. doi: 10.1186/s40360-023-00655-0.

DOI:10.1186/s40360-023-00655-0

PMID:36978114

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10045229/

Abstract

BACKGROUND

The anti-osteosarcoma effects of hydrocortisone and thiram, an inhibitor of type 2 11ß-hydroxysteroid dehydrogenase (11HSD2), have not been reported. The purpose of this study was to investigate the effects of hydrocortisone alone or the combination of hydrocortisone with thiram on osteosarcoma and the molecular mechanism, and determine whether they can be as new therapeutic agents for osteosarcoma.

METHODS

Normal bone cells and osteosarcoma cells were treated with hydrocortisone or thiram alone or in combination. The cell proliferation, migration, cell cycle and apoptosis were detected by using CCK8 assay, wound healing assay, and flow cytometry, respectively. An osteosarcoma mouse model was established. The effect of drugs on osteosarcoma in vivo was assessed by measuring tumor volume. Transcriptome sequencing, bioinformatics analysis, RT-qPCR, Western blotting (WB), enzymelinked immunosorbent assay (ELISA) and siRNA transfection were performed to determine the molecular mechanisms.

RESULTS

Hydrocortisone inhibited the proliferation and migration, and induced apoptosis and cell cycle arrest of osteosarcoma cells in vitro. Hydrocortisone also reduced the volume of osteosarcoma in mice in vivo. Mechanistically, hydrocortisone decreased the levels of Wnt/β-catenin pathway-associated proteins, and induced the expression of glucocorticoid receptor α (GCR), CCAAT enhancer-binding protein β (C/EBP-beta) and 11HSD2, resulting in a hydrocortisone resistance loop. Thiram inhibited the activity of the 11HSD2 enzyme, the combination of thiram and hydrocortisone further enhanced the inhibition of osteosarcoma through Wnt/β-catenin pathway.

CONCLUSIONS

Hydrocortisone inhibits osteosarcoma through the Wnt/β-catenin pathway. Thiram inhibits 11HSD2 enzyme activity, reducing hydrocortisone inactivation and promoting the effect of hydrocortisone through the same pathway.

摘要

背景

氢化可的松和噻二唑啉（一种 2 型 11β-羟类固醇脱氢酶（11HSD2）抑制剂）的抗骨肉瘤作用尚未见报道。本研究旨在探讨氢化可的松单独或联合噻二唑啉对骨肉瘤的作用及其分子机制，并确定其是否可作为骨肉瘤的新治疗药物。

方法

分别用氢化可的松或噻二唑啉单独或联合处理正常骨细胞和骨肉瘤细胞，采用 CCK8 检测、划痕实验和流式细胞术分别检测细胞增殖、迁移、细胞周期和凋亡情况，建立骨肉瘤小鼠模型，通过测量肿瘤体积评估药物对骨肉瘤的体内作用。进行转录组测序、生物信息学分析、RT-qPCR、Western blot（WB）、酶联免疫吸附测定（ELISA）和 siRNA 转染，以确定分子机制。

结果

氢化可的松在体外抑制骨肉瘤细胞的增殖和迁移，并诱导其凋亡和细胞周期停滞。氢化可的松还降低了骨肉瘤小鼠体内的肿瘤体积。机制上，氢化可的松降低了 Wnt/β-catenin 通路相关蛋白的水平，并诱导了糖皮质激素受体α（GCR）、CCAAT 增强子结合蛋白β（C/EBP-beta）和 11HSD2 的表达，从而形成了氢化可的松抵抗环。噻二唑啉抑制 11HSD2 酶的活性，噻二唑啉与氢化可的松联合使用通过 Wnt/β-catenin 通路进一步增强了对骨肉瘤的抑制作用。