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

Alexander Th Wu; Yuan-Chieh Yeh; Yan-Jiun Huang; Ntlotlang Mokgautsi; Bashir Lawal; Tse-Hung Huang

doi:10.1016/j.phymed.2021.153797

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

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

Authors

Alexander Th Wu¹, Yuan-Chieh Yeh², Yan-Jiun Huang³, Ntlotlang Mokgautsi⁴, Bashir Lawal⁴, Tse-Hung Huang⁵

Affiliations

¹ 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.
² 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.
³ Division of Colorectal Surgery, Department of Surgery, Taipei Medical University Hospital, Taipei Medical University, Taipei 11031, Taiwan; Department of Surgery, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan; School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.
⁴ PhD Program for Cancer Molecular Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan; College of Medical Science and Technology, Graduate Institute for Cancer Biology & Drug Discovery, Taipei Medical University, Taipei 11031, Taiwan.
⁵ Department of Traditional Chinese Medicine, Chang Gung Memorial Hospital, Keelung 20401, Taiwan; School of Traditional Chinese Medicine, Chang Gung University, Taoyuan 333, Taiwan; School of Nursing, National Taipei University of Nursing and Health Sciences, Taipei 112, Taiwan; Graduate Institute of Health Industry Technology, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan; Research Center for Chinese Herbal Medicine, Chang Gung University of Science and Technology, Taoyuan 333, Taiwan; Department & Graduate Institute of Chemical Engineering & Graduate Institute of Biochemical Engineering, Ming Chi University of Technology, New Taipei City 243, Taiwan. Electronic address: kchuang@cgmh.org.tw.

PMID: 34802869
DOI: 10.1016/j.phymed.2021.153797

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.

Keywords: Cancer stem cells (CSCs); Cancer-associated fibroblasts (CAFs); Colorectal cancer (CRC); Cyclin-dependent kinase 6 (CDK6); Gamma-mangostin (gMG); Glycogen synthase kinase 3β (GSK3β); miR-26–5p.

MeSH terms

Animals
Carcinogenesis
Cell Line, Tumor
Cell Proliferation
Colon / metabolism
Colorectal Neoplasms* / drug therapy
Colorectal Neoplasms* / genetics
Cyclin-Dependent Kinase 6
Garcinia mangostana* / chemistry
Gene Expression Regulation, Neoplastic
Glycogen Synthase Kinase 3 beta
Humans
Mice
MicroRNAs* / genetics
Molecular Docking Simulation
Wnt Signaling Pathway
Xanthones / pharmacology*
beta Catenin / metabolism

Substances

MicroRNAs
Xanthones
beta Catenin
Glycogen Synthase Kinase 3 beta
CDK6 protein, human
Cyclin-Dependent Kinase 6
mangostin