Journal: BMC Cancer

Article Title: Luminal and basal-like breast cancer cells show increased migration induced by hypoxia, mediated by an autocrine mechanism

doi: 10.1186/1471-2407-11-158

Figure Lengend Snippet: Characterisation of the investigated breast cancer cell lines

Article Snippet: The cell line MCF-7 was derived from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany), the cell lines MDA-MB-231, MDA-MB-435S and MDA-MB-468 were derived from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Amplification

Journal: Oncology Research

Article Title: miR-1193 Suppresses Proliferation and Invasion of Human Breast Cancer Cells Through Directly Targeting IGF2BP2

doi: 10.3727/97818823455816X14760504645779

Figure Lengend Snippet: miR-1193 was significantly downregulated in human breast cancer tissues and cell lines. (A) miR-1193 was downregulated in breast cancer tissues. (B) miR-1193 was decreased in breast cancer cell lines. Breast cancer tissues and paired adjacent noncancerous tissues were sampled from 39 breast cancer patients. The expression levels of miR-1193 were detected in these tissues with real-time qPCR. The expression levels of miR-1193 were detected in human normal breast epithelial cell MCF-10A and human breast cancer cell lines MDA-MB-231, MDA-MB-468, MDA-MB-435, SKBR3, and MCF-7. ** p < 0.01 versus MCF-10A, *** p < 0.001 versus MCF-10A.

Article Snippet: Human normal breast epithelial cell MCF-10A; human breast cancer cell lines, including MDA-MB-231, MDA-MB-468, MDA-MB-435, SKBR3, and MCF-7; and HEK293 cells were purchased from the American Type Culture Collection Company (ATCC; Manassas, VA, USA).

Techniques: Expressing

Journal: Microbial Cell Factories

Article Title: High-yield production of functional soluble single-domain antibodies in the cytoplasm of Escherichia coli

doi: 10.1186/1475-2859-12-97

Figure Lengend Snippet: Analysis of EGFR expression levels and binding affinities of sdAbs to human EGFR-presenting cells. Whole-cell lysates of exponentially growing cells (A431, FaDu, MDA-MB 435S) were prepared and equal amounts of total cellular proteins were separated by SDS-PAGE on 10% polyacrylamide gels. After Western Blot transfer onto PVDF membranes, EGFR and β-actin proteins were detected by incubation with the respective specific antibodies followed by HRP-coupled antibodies and chemiluminescence detection (A) . In vitro specificity of 99m Tc-7C12 and 99m Tc-EG2 on A431 and FaDu cells was investigated after 1 h incubation on ice (B) . Binding of radiolabeled sdAbs was blocked by 40-fold excess of unlabeled Cetuximab. Binding data is expressed as percent of injected dose per mg protein (% ID/mg protein). NB = non-blocked, B = blocked. For in vitro binding studies, two dimensional cultures of A431, FaDu and MDA-MB 435S cells were incubated with increasing concentrations of 99m Tc-7C12 (C) . Total binding was measured in the absence of and nonspecific binding in the presence of 1 mM unlabeled sdAb. Specific binding was calculated as the difference between total and nonspecific binding. Binding studies were repeated twice and representative saturation curves for the EGFR-positive cell lines A431 and FaDu are shown. For the EGFR-negative cell line MDA-MB 435S, no specific binding was observed.

Article Snippet: For binding and uptake studies, three different adherent human tumor cell lines were used: the epidermoid carcinoma cell line A431 (ATCC® Number: CRL-1555), the squamous cell carcinoma cell line FaDu (ATCC® Number: HTB-43), the ductal carcinoma cell line MDA-MB 435S (ATCC® Number: HTB-129).

Techniques: Expressing, Binding Assay, SDS Page, Western Blot, Incubation, In Vitro, Injection

Journal: International Journal of Nanomedicine

Article Title: CD44 + /CD24 − breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold

doi: 10.2147/IJN.S66723

Figure Lengend Snippet: Identification of a CD44 + /CD24 − subpopulation in breast cancer cell lines MCF-10A, MDA-MB-231, BT-549, and MDA-MB-435S by flow cytometry. Notes: Cells in Q4 correspond to CD44 + /CD24 − cells. MDA-MB-435S shows highest proportion of CD44 + /CD24 − antigen phenotype among other cell lines. Abbreviations: FITC, fluorescein isothiocyanate; PE, phycoerythrin.

Article Snippet: The MDA-MB-435S cell line was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Flow Cytometry

Journal: International Journal of Nanomedicine

Article Title: CD44 + /CD24 − breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold

doi: 10.2147/IJN.S66723

Figure Lengend Snippet: The reversion of the malignant phenotype in the morphology of breast cancer cell line MDA-MB-435S in RADA16 compared with Matrigel ® and collagen I scaffolds. Notes: ( A ) AFM images of collagen I, Matrigel, and RADA16 peptide nanofiber scaffolds. Scale bars represent 500 nm. ( B ) Light microscope images (upper) and F-actin and nuclear fluorescence images (lower) of cells encapsulated in collagen I, Matrigel, and RADA16 peptide scaffolds. Three-dimensional cultures were stained for F-actin, and nuclei were counterstained with DAPI. Scale bars represent 100 μm (upper) and 50 μm (lower). ( C ) F-actin and nuclear fluorescence images (upper) and light microscope images (lower) of most common morphology of polarized cell colonies formed in RADA16 scaffold. Scale bars represent 25 μm. ( D ) Encapsulation of cells in different scaffolds using calcein-AM staining for the living cells. Scale bar represents 500 μm. Matrigel ® (BD Biosciences, Two Oak Park, Bedford, MA, USA). Abbreviations: AFM, atomic force microscopy; DAPI, 4′,6-diamidino-2-phenylindole, dihydrochloride; RADA16, COCH 3 -RADARADARADARADA-CONH 2 .

Article Snippet: The MDA-MB-435S cell line was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Light Microscopy, Fluorescence, Staining, Microscopy

Journal: International Journal of Nanomedicine

Article Title: CD44 + /CD24 − breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold

doi: 10.2147/IJN.S66723

Figure Lengend Snippet: The reversion of the malignant phenotype in the proliferation and migration potential of breast cancer cell line MDA-MB-435S in RADA16 compared with Matrigel ® and collagen I scaffolds. Notes: ( A ) Cell density in different scaffolds calculated from DNA measurement after 3-day, 5-day, 7-day, and 9-day culture. ( B ) Percentage of BrdU labeling in cells grown in different scaffolds expressed as the BrdU labeling index from three separate experiments (about 200 cells/experiment) on days 3, 5, 7, and 9. ( C ) Migration potential of MDA-MB-435S cells in Matrigel, collagen I, and RADA16 scaffolds; * P <0.05; ** P <0.001; *** P <0.0001. Matrigel ® (BD Biosciences, Two Oak Park, Bedford, MA, USA). Abbreviations: BrdU, 5-bromo-2′-deoxyuridine; C, collagen; M, Matrigel; R, RADA16; SEM, standard error of the mean; RADA16, COCH 3 -RADARADARADARADA-CONH 2 .

Article Snippet: The MDA-MB-435S cell line was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Migration, Labeling

Journal: International Journal of Nanomedicine

Article Title: CD44 + /CD24 − breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold

doi: 10.2147/IJN.S66723

Figure Lengend Snippet: Cellular characteristics and the localization and expression of signaling proteins in RADA16 cells colonies. Notes: ( A ) Hematoxylin staining of cryostat sections of the Matrigel and RADA16 scaffolds show lumen formation only in RADA16 scaffold. Scale bar represents 25 μm. ( B ) The nuclei of cell colonies stained with DAPI show lumen formation in RADA16 and irregular organization in Matrigel. Scale bar represents 25 μm. ( C ) Immunolocalization of β-catenin (green) with DAPI stained nuclei (blue) in RADA16 and Matrigel scaffolds. The β-catenin is diffusely localized in the cytoplasm, nucleus, and some cell–cell contacts within colonies formed in Matrigel but forms intense localization in the cell–cell junctions in colonies formed in RADA16, indicating similar β-catenin distribution like that of nonmalignant S-1 cells. Scale bar represents 25 μm. ( D ) Western blot analysis of the indicated proteins β-catenin and ICAM-1 of MDA-MB-435S cells in different three-dimensional cultures at 6 days and 9 days. Equal amounts of protein were loaded per lane, and GAPDH was run as a control for equal loading and exposure time. Matrigel ® (BD Biosciences, Two Oak Park, Bedford, MA, USA). Abbreviations: 6 d, 6-day cultures; 9 d, 9-day cultures; C, collagen; DAPI, 4′,6-diamidino-2-phenylindole, dihydrochloride; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ICAM-1, intercellular surface adhesion molecule-1; M, Matrigel; R, RADA16; RADA16, COCH 3 -RADARADARADARADA-CONH 2 .

Article Snippet: The MDA-MB-435S cell line was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Staining, Western Blot

Journal: International Journal of Nanomedicine

Article Title: CD44 + /CD24 − breast cancer cells exhibit phenotypic reversion in three-dimensional self-assembling peptide RADA16 nanofiber scaffold

doi: 10.2147/IJN.S66723

Figure Lengend Snippet: The reversion phenotype of MDA-MB-435S cells was inhibited by blocking NF-κB signaling by PDTC. Notes: ( A ) Western blot analysis of ICAM-1 of MDA-MB-435S cells in different culture groups. Equal amounts of protein were loaded per lane, and GAPDH was run as a control for equal loading and exposure time. The expression of ICAM-1 of cells in RADA16 was significantly downregulated by PDTC (* P <0.05). ( B ) Light microscope images and F-actin and nuclear fluorescence images of cells in PDTC-treated RADA16 group. Three-dimensional cultures were stained for F-actin, and nuclei were counterstained with DAPI. Scale bars represent 100 μm (left) and 50 μm (right). Abbreviations: 2D, two dimensional group; DAPI, 4′,6-diamidino-2-phenylindole, dihydrochloride; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ICAM-1, intercellular surface adhesion molecule-1; PDTC, pyrrolidine dithiocarbamate; R, RADA16 group; Rp, RADA16 + PDTC group; RADA16, COCH 3 -RADARADAR-ADARADA-CONH 2 .

Article Snippet: The MDA-MB-435S cell line was obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Blocking Assay, Western Blot, Expressing, Light Microscopy, Fluorescence, Staining