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    Adam Cerise et al
    Supplemental Data
    Supplemental Figure 1 Change of Body Weight of Mice in
    Various Treatment Group (n [ 8 and Repeated 3 Times) Described in Figure 4
    Abbreviation: Act D ¼ actinomycin D.
    Supplemental Figure 2 In Vitro Activity of Oxaliplatin in
    Combination of LMB-100 on SW-48 Cell Line (Repeated Twice)
    Abbreviation: PBS ¼ phosphate-buffered saline.
    Contents lists available at ScienceDirect
    journal homepage:
    Original article
    Anti-metastasis activity of curcumin against breast cancer via the inhibition T of stem cell-like properties and EMT
    Hu Chenxiaa,1, Li Mengjiea,1, Guo Tingtinga, Wang Shaoxib, Huang Weipingc, Yang Kea, Liao Zhiweid, Wang Jianc, Zhang Fengxuec, , Wang Hongqic, a School of Pharmaceutical Science, Guangzhou University of Chinese Medicine, Guangzhou, China b School of Software & Microelectronics, Northwestern Polytechnical University, Xi'an, China c The Research Center of Basic Integrative Medicine, Guangzhou University of Chinese Medicine, Guangzhou, China d Cancer Center of Guangzhou Medical University, Guangzhou, China
    Breast cancer
    Epithelial-mesenchymal transition
    Breast cancer stem-like cells
    Background: Curcumin is a polyphenolic compound with potent chemopreventive and anti-cancer efficacy. Purpose: To explore the potential anti-metastasis efficacy of curcumin in breast cancer stem-like Infliximab (BCSCs), which are increasingly considered to be the origin of the recurrence and metastasis of breast cancer. Methods: A CCK8 assay was performed to evaluate cell viability, and a colony formation assay was conducted to determine cell proliferation in MCF-7 and MDA-MB-231 adherent cells. Transwell and wound healing assays were used to detect the effect of curcumin on cell migration and invasion in MDA-MB-231 cells. Mammospheres were cultured with serum free medium (SFM) for three generations and the BCSC surface marker CD44+CD24−/low subpopulation was measured by flow cytometry. Mammosphere formation and differentiation abilities were determined after cell treatment with curcumin. Then, a reverse transcription-quantitative poly-merase chain reaction assay was conducted to detect the relative mRNA level of epithelial-mesenchymal tran-sition (EMT) marker genes and western blot analysis was performed to determine the protein expression of stem cell genes in mammospheres treated with curcumin.
    Results: Curcumin exhibited anti-proliferative and colony formation inhibiting activities in both the MCF-7 and MDA-MB-231 cell lines. It also suppressed the migration and invasion of MDA-MB-231 cells. The CD44 +CD24−/low subpopulation was larger in mammospheres when MCF-7 and MDA-MB-231 adherent cells were cultured with SFM. Further studies revealed that curcumin inhibited mammosphere formation and dif-ferentiation abilities. Moreover, curcumin down-regulated the mRNA expression of Vimentin, Fibronectin, and β-catenin, and up-regulated E-cadherin mRNA expression levels. Western blot analysis demonstrated that cur-cumin decreased the protein expression of stem cell genes including Oct4, Nanog and Sox2.
    Conclusion: The results of the present study suggest that the inhibitor effects of curcumin on breast cancer cells may be related to resistance to cancer stem-like characters and the EMT process. These data indicate that cur-cumin could function as a type of anti-metastasis agent for breast cancer.
    Introduction accounting for over 8 million deaths every year. Breast cancer is
    Cancer is the second leading cause of human mortality worldwide, thought to account for ∼30% of all estimated new cancer cases and
    14% of estimated cancer deaths among women in the United States
    Abbreviations: AML, acute myeloid leukemia; ATCC, American type culture collection; BCA, bicinchoninic acid; BCSCs, breast cancer stem-like cells; bFGF, basic fibroblast growth factor; CCK8, cell counting Kit-8; CSCs, cancer stem cells; DMEM, Dulbecco's modified eagle's medium; DMSO, dimethyl sulfoxide; EDTA, ethylene diamine tetraacetic acid; EGF, epidermal growth factor; EMT, epithelial-mesenchymal transition; FBS, fetal bovine serum; FCM, flow cytometry; FITC, fluorescein isothiocyanate; PBS, phosphate buffer saline; PE, phycoerythrin; PMSF, phenylmethylsulfonyl fluoride; PVDF, polyvinylidene difluoride; RIPA, radio-immunoprecipitation assay; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel elec-trophoresis; SFM, serum free medium; SSM, serum-supplemented medium; TBST, tris-buffered saline tween-20