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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.
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Phytomedicine
journal homepage: www.elsevier.com/locate/phymed
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
Keywords:
Curcumin
Breast cancer
Epithelial-mesenchymal transition
Breast cancer stem-like cells
Metastasis
Mammospheres
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 Ezatiostat (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