Ovatodiolide, isolated from Anisomeles indica, suppresses bladder carcinogenesis through suppression of mTOR/β-catenin/CDK6 and exosomal miR-21 derived from M2 tumor-associated macrophages

Toxicol Appl Pharmacol. 2020 Aug 15:401:115109. doi: 10.1016/j.taap.2020.115109. Epub 2020 Jun 13.

Abstract

Bladder cancer (BCa) is the fourth leading cause of cancer deaths worldwide due to its aggressiveness and resistance against therapies. Intricate interactions between cancer cells and the tumor microenvironment (TME) are essential for both disease progression and regression. Thus, interrupting molecular communications within the TME could potentially provide improved therapeutic efficacies. M2-polarized tumor-associated macrophages (M2 TAMs) were shown to contribute to BCa progression and drug resistance. We attempted to provide evidence for ovatodiolide (OV) as a potential therapeutic agent that targets both TME and BCa cells. First, tumor-suppressing functions of OV were determined by cell viability, colony, and tumor-sphere formation assays using a coculture system composed of M2 TAMs/BCa cells. Subsequently, we demonstrated that extracellular vesicles (EVs) isolated from M2 TAMs containing oncomiR-21 and mRNAs, including Akt, STAT3, mTOR, and β-catenin, promoted cisplatin (CDDP) resistance, migration, and tumor-sphere generation in BCa cells, through increasing CDK6, mTOR, STAT3, and β-catenin expression. OV treatment also prevented M2 polarization and reduced EV cargos from M2 TAMs. Finally, in vivo data demonstrated that OV treatment overcame CDDP resistance. OV only and the OV + CDDP combination both resulted in significant reductions in mTOR, β-catenin, CDK6, and miR-21 expression in tumor samples and EVs isolated from serum. Collectively, we demonstrated that M2 TAMs induced malignant properties in BCa cells, in part via oncogenic EVs. OV treatment prevented M2 TAM polarization, reduced EV cargos derived from M2 TAMs, and suppressed β-catenin/mTOR/CDK6 signaling. These findings provide preclinical evidence for OV as a single or adjuvant agent for treating drug-resistant BCa.

Keywords: Bladder Cancer; Extracellular Vesicle; M2 Tumor-Associated Macrophage; Ovatodiolide; mTOR/CDK6/β-Catenin Signaling, Therapeutic Development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carcinogenesis / drug effects
  • Carcinogenesis / metabolism
  • Cell Line, Tumor
  • Coculture Techniques
  • Cyclin-Dependent Kinase 6 / antagonists & inhibitors
  • Cyclin-Dependent Kinase 6 / metabolism*
  • Diterpenes / isolation & purification
  • Diterpenes / pharmacology
  • Diterpenes / therapeutic use*
  • Dose-Response Relationship, Drug
  • Exosomes / drug effects
  • Exosomes / metabolism
  • Exosomes / pathology
  • Female
  • Humans
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Macrophages / pathology
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • MicroRNAs / antagonists & inhibitors
  • MicroRNAs / metabolism*
  • Plants, Medicinal
  • TOR Serine-Threonine Kinases / antagonists & inhibitors
  • TOR Serine-Threonine Kinases / metabolism*
  • Urinary Bladder Neoplasms / drug therapy
  • Urinary Bladder Neoplasms / metabolism*
  • Urinary Bladder Neoplasms / pathology
  • beta Catenin / antagonists & inhibitors
  • beta Catenin / metabolism*

Substances

  • CTNNB1 protein, human
  • Diterpenes
  • MIRN21 microRNA, human
  • MicroRNAs
  • beta Catenin
  • ovatodiolide
  • MTOR protein, human
  • TOR Serine-Threonine Kinases
  • CDK6 protein, human
  • Cyclin-Dependent Kinase 6