Changes in Tumor Stem Cell Markers and Epithelial-Mesenchymal Transition Markers in Nonluminal Breast Cancer after Neoadjuvant Chemotherapy and Their Correlation with Contrast-Enhanced Ultrasound

Nonluminal breast cancer has high early metastasis and treatment resistance, and neoadjuvant chemotherapy (NAC) is needed. The presence of cancer stem cells (CSC) and epithelial-mesenchymal transition (EMT) leads to poor prognosis. This study investigated the changes in CSC markers and EMT markers after NAC in nonluminal breast cancer and their correlation with contrast-enhanced ultrasound (CEUS) features and chemotherapy efficacy. Before NAC, the range of nonluminal breast cancer on CEUS was larger than that of two-dimensional ultrasound, but after NAC, it was significantly smaller than that of two-dimensional ultrasound and closer to the postoperative pathological size. After NAC, the enlarged lesions and perfusion defects were significantly less than those before NAC. The time-intensity curve showed the characteristics of slow-in, low enhancement, and low perfusion. Nonluminal breast cancer downregulated the expression of CSC markers and EMT markers after NAC, but the epithelial phenotype of nonluminal breast cancer with good response to chemotherapy was upregulated. In nonluminal breast cancer with poor response to chemotherapy, markers of CSC and EMT were highly expressed before chemotherapy. In conclusion, CEUS is better than conventional ultrasound in estimating NAC efficacy in this mode. CEUS can also predict the prognosis of nonluminal breast cancer before NAC with the characteristics of enhanced enlargement and perfusion defects. The contrast-enhanced time-intensity curve of lesions with relatively poor blood supply may have more CSC and EMT characteristics.

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P53 Silencing and Mammosphere Formation in Breast Cancer Cells Harbouring P53 Gain-of-Function Mutations Trigger Epithelial-Mesenchymal Transition

Journal of Global Oncology ◽

10.1200/jgo.18.81200 ◽

2018 ◽

Vol 4 (Supplement 2) ◽

pp. 201s-201s

Author(s):

Z.Y. Yee ◽

C.L. Lim ◽

F.L. Felicia Chung ◽

C.O. Leong

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Cell Lines ◽

Breast Cancer Cells ◽

Epithelial Mesenchymal Transition ◽

P53 Gene ◽

Mesenchymal Transition ◽

Cell Markers ◽

Emt Markers ◽

E Cadherin

Background: Mutations in p53 gene are observed in ∼50% of all human cancers. In breast cancer alone, 12%-32% of luminal, 84% of basal-like and 75% of HER-2 expressing tumors have apparent p53 mutations. Tumor cells undergo epithelial-mesenchymal transition (EMT) to metastasise from primary sites to form secondary tumors at distant regions of the body. EMT is a complex biologic phenomenon which governs the transition of cancer cells with epithelial characteristics to mesenchymal traits, gaining new properties such as aggressiveness and invasiveness. Recent studies revealed that mutations in the p53 gene can give rise to alternate functional phenotypes leading to tumor initiation and progression. Aim: The aim of this study is to develop a robust human breast cancer cellular model to investigate p53 gain-of-function (GOF) mutations and EMT as well as evaluating the EMT phenotype associated with these mutations. Methods: Two breast cancer cell lines, namely MDA-MB-468 and HCC38 carrying the R273H and R273L missense mutations, respectively, were subjected to p53 knockdown using shRNA directed against p53 gene through lentiviral vector transduction. The transduced cells were then harvested for Western blotting to evaluate the protein expression of EMT markers which includes E-cadherin, SNAIL, ZEB1 and vimentin compared with the nontransduced control cells. Subsequently, both cell lines were subjected to mammosphere generation and redifferentiation to determine the basal expression of the EMT markers. Results: Silencing of p53 using siRNA in MDA-MB-468 and HCC38 downregulated E-cadherin expressions but upregulated vimentin levels. Furthermore, E-cadherin levels reduced significantly after conversion from adherent parental cells to mammospheres, but rebound upon redifferentiation. Conversely, vimentin was upregulated in mammospheres as compared with the parental and redifferentiated groups. Conclusion: p53 knockdown in breast cancer cells harboring R273H and R273L mutations favor vimentin expression but not E-cadherin, suggesting that p53-GOF mutants are involved in EMT and the development of metastatic tumors. The mammosphere model accurately recapitulates cell plasticity between epithelial and mesenchymal states, as evidenced by the expression of mesenchymal cell markers in the mammospheres, and epithelial cell markers in adherent and redifferentiated cells.

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Contrast-enhanced ultrasound for evaluating the response of breast cancer to neoadjuvant chemotherapy: Time-intensity curve analysis and texture analysis

Annals of Oncology ◽

10.1093/annonc/mdw365.86 ◽

2016 ◽

Vol 27 ◽

pp. vi96

Author(s):

C. Yuan ◽

L. Tang ◽

Q. Zhang ◽

W. Jia ◽

M. Chen

Keyword(s):

Breast Cancer ◽

Neoadjuvant Chemotherapy ◽

Texture Analysis ◽

Curve Analysis ◽

Contrast Enhanced Ultrasound ◽

Intensity Curve ◽

Time Intensity Curve ◽

Contrast Enhanced

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Sulforaphane Modulates Cell Migration and Expression of βCatenin and Epithelial Mesenchymal Transition Markers in Breast Cancer Cells

Iranian Journal of Public Health ◽

10.18502/ijph.v49i1.3054 ◽

2020 ◽

Author(s):

Mehdi BAGHERI ◽

Mozhgan FAZLI ◽

Sara SAEEDNIA ◽

Majid GHOLAMI KHARANAGH ◽

Naghmeh AHMADIANKIA

Keyword(s):

Breast Cancer ◽

Cell Migration ◽

Epithelial Mesenchymal Transition ◽

Flow Cytometric Analysis ◽

Drug Candidate ◽

Control Group ◽

Mesenchymal Transition ◽

Time Points ◽

Induced Apoptosis ◽

Emt Markers

Background: We aimed to assess the effect of sulforaphane (SFN) on breast cancer cell migration and also its effect on the expression of epithelial mesenchymal transition (EMT) markers and β-catenin. Methods: This study was performed in Shahroud University of Medical Sciences, Shahroud, Iran from 2017- 2018. In this experimental study, MDA-MB-231 cells were treated with different concentrations of SFN (5, 10, 20, 30 and 40 μM) at different time points of 24, 48, and 72 h. The control group was untreated cells. The inhibitory effects of different concentrations of SFN on cell migration at different time points were evaluated using scratch assay. Moreover, apoptosis was assessed by using flow cytometric analysis. The expression of βcatenin and EMT markers of ZEB1, fibronectin, and claudin-1 were determined by real-time PCR. Western blotting analysis of β-catenin was applied to determine its changes after SFN treatment. Results: SFN markedly inhibited the migration of cells at concentrations of 10, 20, 30, and 40µM after 24, 48, and 72 h. At relatively, high concentrations (30, 40µM), SFN induced apoptosis. Moreover, SFN reduced the gene expression of ZEB1, fibronectin, and claudin-1 after 72 h. The expression of β-catenin revealed a timedependent decrease at the concentration of 40 µM SFN. Conclusion: Downregulation of EMT markers and β-catenin showed accordance with the inhibition of migration. SFN could be a promising drug candidate to reduce metastasis in breast cancer.

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