从藤黄属植物中分离得到的γ-倒捻子素通过下调 GSK3β/β-连环蛋白/CDK6 癌症干细胞通路抑制结肠癌变和干性。

Gamma-mangostin isolated from garcinia mangostana suppresses colon carcinogenesis and stemness by downregulating the GSK3β/β-catenin/CDK6 cancer stem pathway.

机构信息

The PhD Program of Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan; International PhD Program for Translational Science, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan; College of Medical Science and Technology, Graduate Institute of Biomedical Informatics, Taipei Medical University, Taipei 11031, Taiwan; Clinical Research Center, Taipei Medical University Hospital, Taipei Medical University, Taipei 11031, Taiwan; Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei 114, Taiwan.

Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung, 20401, Taiwan; Program in Molecular Medicine, School of Life Sciences, National Yang Ming University, Taipei 11221, Taiwan.

出版信息

Phytomedicine. 2022 Jan;95:153797. doi: 10.1016/j.phymed.2021.153797. Epub 2021 Oct 21.

DOI:10.1016/j.phymed.2021.153797

PMID:34802869

Abstract

BACKGROUND

Despite advances in chemotherapies and targeted drugs, colorectal cancer (CRC) remains challenging to treat due to drug resistance. Emerging evidence indicates that cancer-associated fibroblasts (CAFs) facilitate the generation of cancer stem-like cells (CSCs) and drug resistance. Glycogen synthase kinase-3 (GSK) associated signaling pathways have been implicated in the generation of CSCs and represent a target for therapeutics development.

HYPOTHESIS

Gamma-mangostin (gMG) isolated from Garcinia mangostana was evaluated for its ability to downregulate GSK3β-associated signaling in CRC cells and overcome CAF-induced 5-fluorouracil resistance and CSC generation.

METHODS

Bioinformatics analysis, in silico molecular docking, in vitro assays, including cell viability tests, colony- and tumor sphere-formation assays, transwell migration assays, ELISA, SDS-PAGE, Western blotting, miR expression, qPCR, and flow cytometry, as well as in vivo mouse xenograft models were used to evaluate the antitumor effects of gMG.

RESULTS

Bioinformatics analyses indicated that GSK3β/CDK6/β-catenin mRNA signature was significantly higher in colon cancer patients. Additional algorithms predicted a higher miR-26b level was associated with significantly higher survival in CRC patients and GSK3β and CDK6 as targets of miR-26b-5p. To validate these findings in vitro, we showed that CAF-cocultured CRC cells expressed an increased expression of GSK3β, β-catenin, CDK6, and NF-κB. Therapeutically, we demonstrated that gMG treatment suppressed GSK3β-associated signaling pathways while concomitantly increased the miR-26b-5p level. Using a xenograft mouse model of CAFs cocultured HCT116 tumorspheres, we showed that gMG treatment reduced tumor growth and overcame CAF-induced 5-fluorouracil resistance.

CONCLUSIONS

Pharmacological intervention with gMG suppressed CRC carcinogenesis and stemness via downregulating GSK3/β-catenin/CDK6 and upregulating the miR-26b-5p tumor suppressor. Thus, gMG represents a potential new CRC therapeutic agent and warrants further investigation.

摘要

背景

尽管化疗和靶向药物有了进步，但由于耐药性，结直肠癌（CRC）的治疗仍然具有挑战性。新出现的证据表明，癌相关成纤维细胞（CAFs）促进了癌症干细胞样细胞（CSCs）的产生和耐药性。糖原合成酶激酶-3（GSK）相关信号通路与 CSCs 的产生有关，是治疗药物开发的靶点。

假说

从藤黄属植物中分离出的γ-倒捻子素（gMG），其能够下调 CRC 细胞中 GSK3β 相关信号，并克服 CAF 诱导的 5-氟尿嘧啶耐药和 CSC 生成。

方法

采用生物信息学分析、计算机分子对接、体外试验，包括细胞活力试验、集落和肿瘤球形成试验、Transwell 迁移试验、ELISA、SDS-PAGE、Western blot、miR 表达、qPCR 和流式细胞术，以及体内小鼠异种移植模型，评估 gMG 的抗肿瘤作用。

结果

生物信息学分析表明，结肠癌患者的 GSK3β/CDK6/β-catenin mRNA 特征明显更高。其他算法预测 miR-26b 水平较高与 CRC 患者的生存率显著提高有关，GSK3β 和 CDK6 是 miR-26b-5p 的靶点。为了在体外验证这些发现，我们表明 CAF 共培养的 CRC 细胞表达了更高的 GSK3β、β-catenin、CDK6 和 NF-κB 表达。治疗上，我们证明 gMG 治疗抑制了 GSK3β 相关信号通路，同时增加了 miR-26b-5p 的水平。使用 CAFs 共培养 HCT116 肿瘤球的异种移植小鼠模型，我们表明 gMG 治疗减少了肿瘤生长并克服了 CAF 诱导的 5-氟尿嘧啶耐药性。