scholarly journals Revealing Glycoproteins in the Secretome of MCF-7 Human Breast Cancer Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Aik-Aun Tan ◽  
Wai-Mei Phang ◽  
Subash C. B. Gopinath ◽  
Onn H. Hashim ◽  
Lik Voon Kiew ◽  
...  

Breast cancer is one of the major issues in the field of oncology, reported with a higher prevalence rate in women worldwide. In attempt to reveal the potential biomarkers for breast cancer, the findings of differentially glycosylated haptoglobin and osteonectin in previous study have drawn our attention towards glycoproteins of secretome from the MCF-7 cancer cell line. In the present study, further analyses were performed on the medium of MCF-7 cells by subjecting it to two-dimensional analyses followed by image analysis in contrast to the medium of human mammary epithelial cells (HMEpC) as a negative control. Carboxypeptidase A4 (CPA4), alpha-1-antitrypsin (AAT), haptoglobin (HP), and HSC70 were detected in the medium of MCF-7, while only CPA4 and osteonectin (ON) were detected in HMEpC medium. In addition, CPA4 was detected as upregulated in the MCF-7 medium. Further analysis by lectin showed that CPA4, AAT, HP, and HSC70 were secreted as N-glycan in the medium of MCF-7, with HP also showing differentially N-glycosylated isoforms. For the HMEpC, only CPA4 was detected as N-glycan. No O-glycan was detected in the medium of HMEpC but MCF-7 expressed O-glycosylated CPA4 and HSC70. All these revealed that glycoproteins could be used as glycan-based biomarkers for the prognosis of breast cancer.

2021 ◽  
Vol 22 (2) ◽  
pp. 800
Author(s):  
Keita Iwasaki ◽  
Asuka Araki ◽  
C Murali Krishna ◽  
Riruke Maruyama ◽  
Tatsuyuki Yamamoto ◽  
...  

Raman spectroscopy (RS), a non-invasive and label-free method, has been suggested to improve accuracy of cytological and even histopathological diagnosis. To our knowledge, this novel technique tends to be employed without concrete knowledge of molecular changes in cells. Therefore, identification of Raman spectral markers for objective diagnosis is necessary for universal adoption of RS. As a model study, we investigated human mammary epithelial cells (HMEpC) and breast cancer cells (MCF-7) by RS and employed various multivariate analyses (MA) including principal components analysis (PCA), linear discriminant analysis (LDA), and support vector machine (SVM) to estimate diagnostic accuracy. Furthermore, to elucidate the underlying molecular changes in cancer cells, we utilized multivariate curve resolution analysis–alternating least squares (MCR-ALS) with non-negative constraints to extract physically meaningful spectra from complex cellular data. Unsupervised PCA and supervised MA, such as LDA and SVM, classified HMEpC and MCF-7 fairly well with high accuracy but without revealing molecular basis. Employing MCR-ALS analysis we identified five pure biomolecular spectra comprising DNA, proteins and three independent unsaturated lipid components. Relative abundance of lipid 1 seems to be strictly regulated between the two groups of cells and could be the basis for excellent discrimination by chemometrics-assisted RS. It was unambiguously assigned to linoleate rich glyceride and therefore serves as a Raman spectral marker for reliable diagnosis. This study successfully identified Raman spectral markers and demonstrated the potential of RS to become an excellent cytodiagnostic tool that can both accurately and objectively discriminates breast cancer from normal cells.


2004 ◽  
Vol 24 (12) ◽  
pp. 5548-5564 ◽  
Author(s):  
Jason D. Prescott ◽  
Karen S. N. Koto ◽  
Meenakshi Singh ◽  
Arthur Gutierrez-Hartmann

ABSTRACT Several different transcription factors, including estrogen receptor, progesterone receptor, and ETS family members, have been implicated in human breast cancer, indicating that transcription factor-induced alterations in gene expression underlie mammary cell transformation. ESE-1 is an epithelium-specific ETS transcription factor that contains two distinguishing domains, a serine- and aspartic acid-rich (SAR) domain and an AT hook domain. ESE-1 is abundantly expressed in human breast cancer and trans-activates epithelium-specific gene promoters in transient transfection assays. While it has been presumed that ETS factors transform mammary epithelial cells via their nuclear transcriptional functions, here we show (i) that ESE-1 protein is cytoplasmic in human breast cancer cells; (ii) that stably expressed green fluorescent protein-ESE-1 transforms MCF-12A human mammary epithelial cells; and (iii) that the ESE-1 SAR domain, acting in the cytoplasm, is necessary and sufficient to mediate this transformation. Deletion of transcriptional regulatory or nuclear localization domains does not impair ESE-1-mediated transformation, whereas fusing the simian virus 40 T-antigen nuclear localization signal to various ESE-1 constructs, including the SAR domain alone, inhibits their transforming capacity. Finally, we show that the nuclear localization of ESE-1 protein induces apoptosis in nontransformed mammary epithelial cells via a transcription-dependent mechanism. Together, our studies reveal two distinct ESE-1 functions, apoptosis and transformation, where the ESE-1 transcription activation domain contributes to apoptosis and the SAR domain mediates transformation via a novel nonnuclear, nontranscriptional mechanism. These studies not only describe a unique ETS factor transformation mechanism but also establish a new paradigm for cell transformation in general.


2020 ◽  
Vol 20 (10) ◽  
pp. 6561-6567 ◽  
Author(s):  
Bo Zheng ◽  
Minghua Xue ◽  
Xinyi Zhang ◽  
Ning Tian ◽  
Dongmei Wang

Objective: This study aimed to determine the effects of dimer captosuccinic acid-coated Fe3O4 (super paramagnetic) nanoparticles (NP) on 2-deoxy-d-glucose in targeted cancer cells with high rates of glucose metabolism. Methods: We prepared Fe3O4@DMSA NP and 2-DG-conjugated Fe3O4@DMSA NP, γ-FE, O, and @DMSA-DG NP. Glucose consumption in MDA-MB-231 and MCF-7 breast cancer cells was determined using γ-Fe2O3@DMSA NP or Fe3O4@DMSA-DG NP, and absorption was tested using Prussian blue staining, ultraviolet colorimetry, and magnetic resonance imaging. Results: Glucose consumption was the highest in MDA-MB-231, and the lowest in human mammary epithelial cells (HMEPiC). The significant uptake of Fe2O3@DMSA-DG NP by MDA-MB-231 and MCF-7 cells within two hours was inhibited by glucose. The uptake of Fe3O4@DMSA-DG NP was significantly higher in MDA-MB-231 than in MCF-7 cells, whereas Fe3O4@DMSA NP was not obviously uptaken by either cell line. Absorption was also not evident in HMEPiC incubated with Fe3O4@DMSA-DG NP and Fe3O4@DMSA NP. Conclusions: The tumor targeting efficacy of 2-DG coated Fe3O4@DMSA NP was improved over Fe3O4,@DMSA NP in cancer cells with high rates of glucose metabolism.


BMC Cancer ◽  
2010 ◽  
Vol 10 (1) ◽  
Author(s):  
Joanna M Poczobutt ◽  
John Tentler ◽  
Xian Lu ◽  
Pepper J Schedin ◽  
Arthur Gutierrez-Hartmann

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