title: Sulforaphane suppresses EMT and metastasis in human lung cancer through miR-616-5p-mediated GSK3β/β-catenin signaling pathways.
abstract: Sulforaphane is a common antioxidant selectively abundant in cruciferous plants, which exhibits effective anti-cancer actions in control of tumorigenesis or progression of various cancers. A recent study has shown that sulforaphane attenuates the EGFR signaling pathway in non-small cell lung cancer (NSCLC), suggesting its potential anti-metastatic effects. In this study we assessed the involvement of sulforaphane and miR-616-5p in epithelial-mesenchymal transition (EMT) and NSCLC metastasis. Sulforaphane suppressed the cell proliferation in human NSCLC cell lines H1299, 95C and 95D with IC50values of 9.52±1.23, 9.04±1.90 and 17.35±2.03 μmol/L, respectively. At low concentrations (1-5 μmol/L), sulforaphane dose-dependently inhibited the migration and invasion of 95D and H1299 cells with relatively high metastatic potential. The anti-metastatic action of sulforaphane was confirmed in 95D and H1299 cell xenografts in vivo. In fresh NSCLC tissue samples from 179 patients, miR-616-5p levels were upregulated in late-stage NSCLCs, and strongly correlated with risk of NSCLC recurrence and metastasis. Consistent with the clinic observation, miR-616-5p levels in the 3 NSCLC cell lines were correlated with their metastatic ability, and were decreased by sulforaphane treatment. Silencing miR-616-5p markedly suppressed the migration and invasion of 95D cells in vitro and NSCLC metastasis in vivo. Further studies revealed that miR-616-5p directly targeted GSK3β and decreased its expression, whereas sulforaphane decreased miR-616-5p levels by histone modification, and followed by inactivation of the GSK3β/β-catenin signaling pathway and inhibition of EMT, which was characterized by loss of epithelial markers and acquisition of a mesenchymal phenotype in NSCLC cells. Our findings suggest that sulforaphane is a potential adjuvant chemotherapeutic agent for the prevention of NSCLC recurrence and metastasis, and miR-616-5p can be clinically utilized as a biomarker or therapeutic target to inhibit metastasis.
authors.affiliation: Department of Respiratory Medicine, the First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China.
authors.forename: Da-Xuan
authors.initials: DX
authors.lastname: Wang
authors.affiliation: Department of Oncology, Traditional Chinese Medicine-Integrated Hospital of Southern Medical University, Guangzhou 510310, China.
authors.forename: Yu-Jiao
authors.initials: YJ
authors.lastname: Zou
authors.affiliation: Department of Respiratory Medicine, the First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China.
authors.forename: Xi-Bin
authors.initials: XB
authors.lastname: Zhuang
authors.affiliation: Department of Thoracic Surgery, Fuzhou Pulmonary Hospital, Fujian Medical University, Fuzhou 350008, China.
authors.forename: Shu-Xing
authors.initials: SX
authors.lastname: Chen
authors.affiliation: Department of Thoracic Surgery, Fuzhou Pulmonary Hospital, Fujian Medical University, Fuzhou 350008, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Department of Respiratory Medicine, the First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China.
authors.forename: Wen-Lan
authors.initials: WL
authors.lastname: Li
authors.affiliation: College of Basic Medicine, Fujian Medical University, Fuzhou 350108, China.
authors.forename: Jun-Jin
authors.initials: JJ
authors.lastname: Lin
authors.affiliation: Department of Pharmacy, the First Hospital of Quanzhou Affiliated to Fujian Medical University, Quanzhou 362000, China.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Lin
mesh_terms: D000818:Animals; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism*; D006801:Humans; D017879:Isothiocyanates / Q000494:pharmacology / Q000627:therapeutic use*; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D009362:Neoplasm Metastasis / Q000188:drug therapy*; D009364:Neoplasm Recurrence, Local / Q000188:drug therapy; D015398:Signal Transduction / Q000187:drug effects*; D013454:Sulfoxides; D023041:Xenograft Model Antitumor Assays; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D017879:Isothiocyanates; C584055:MIRN616 microRNA, human; D035683:MicroRNAs; D013454:Sulfoxides; D051176:beta Catenin; D000071679:Glycogen Synthase Kinase 3 beta; C016766:sulforaphane