scholarly journals Front Cover: Removal of Serum Lipids and Lipid‐Derived Metabolites to Investigate Breast Cancer Cell Biology

PROTEOMICS ◽  
2019 ◽  
Vol 19 (18) ◽  
pp. 1970161
Author(s):  
Viktor Brovkovych ◽  
Alyssa Aldrich ◽  
Nasi Li ◽  
G. Ekin Atilla‐Gokcumen ◽  
Jonna Frasor
PROTEOMICS ◽  
2019 ◽  
Vol 19 (18) ◽  
pp. 1800370 ◽  
Author(s):  
Viktor Brovkovych ◽  
Alyssa Aldrich ◽  
Nasi Li ◽  
G. Ekin Atilla‐Gokcumen ◽  
Jonna Frasor

2019 ◽  
Vol 2 (2) ◽  
pp. 29 ◽  
Author(s):  
Carla Luis ◽  
Yuselis Castaño-Guerrero ◽  
Raquel Soares ◽  
Goreti Sales ◽  
Rúben Fernandes

Doxorubicin (DOXO) is an adjuvant chemotherapy agent and is also commonly used in cell biology research. Cytotoxic assays in cell culture are frequently used in order to stablish drug concentrations that are useful for controlling cell proliferation. One common cytotoxic method used is 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT). Our present research aims to support future studies in engaging MTT assay using DOXO that exhibits a strong red coloration and fluorescence, and so it is assumed that DOXO may interfere with commonly used colorimetric assays such as MTT. The interference of DOXO in the MTT determination was evaluated in a Breast Cancer cell line Michigan Cancer Foundation-7 (MCF-7). The interference was evaluated by means of spectroscopic methods in particular spectrophometry and fluorescence spectroscopy of MTT and DOXO. We postulate that the medium and the MTT reagent itself can interfere on the metabolic activity method, so in order to achieve better results, DMEM was replaced by a neutral buffer like Phosphate-buffered saline (PBS). This protocol may be extremely useful in future studies involving DOXO.


2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e22055-e22055
Author(s):  
L. Putignani ◽  
S. Raffa ◽  
R. Pescosolido ◽  
F. Signore ◽  
D. Menichella ◽  
...  

e22055 Background: Mitochondriopathy has been recently rekindled as new cancer theory. We report on structural damage of breast-infiltrating ductal carcinoma (IDC) mitochondria characterised by reduced expression levels of the oxidative phosphorylation system (OXPHOS). Methods: Mitochondria from HMC-1 (human mammary carcinoma) and HMEC (human mammary epithelial cell) cultures, traced by Mitotracker, were assayed for OXPHOS expression levels using cryo-immunoelectron microscopy (CIEM) quantitative labelling and fluorescence immunolabelling on unfractionated HMC-1 and HEMC cells. Convolution degeneration was established by transmission electron microscopy (TEM). Twenty different cell sections for both HMC-1 and HEMC cells, including 65 and 72 mitochondria, respectively, were randomly recorded and quantitatively analyzed for the percentage of area occupied by intact cristae to provide a grading of mitochondrial damage (cristae loss index). Results: Depressed expression levels were detected for all HMC-1 OXPHOS complexes by CIEM. Normalized labelling density (HEMC/HMC-1), expressed as colloidal gold particles/mitochondrial area (ρ) provided the following values: 1.77 for the NADH-ubiquinone oxidoreductase complex I NDUFS3; 1.86 for the succinate- dehydrogenase complex II SDH-B protein; 1.63 for the ubiquinol cytochrome c reductase complex III UQCRC2; 4.88 and 1.58 for the cytochrome-oxidase complex IV (CO) subunit I and IV, respectively; 2.70 for the ATP-synthase complex V F1β protein. Fluorescence immunolabelling confirmed CIEM quantitative data. MitoTracker's co-staining showed altered membrane potential and permeability. Injury grading was categorised assigning three levels of morphological damage: i) severe, ii) moderate, iii) slight, corresponding to 0 % (6.2 % and 1.4 % for HMC-1 and HMEC, respectively), 1–50 % (21.5 % and 2.8 % for HMC-1 and HMEC, respectively) and 51–75 % (44.6 % and 15.3 % for HMC-1 and HMEC, respectively) of area occupied by intact cristae (p<0.0001, χ2Test). The entire HMC-1 mitochondrial damage resulted 3.7 times higher than that observed for HMEC cells (72.3HMC-1 %/19.5HMEC %). Conclusions: New phenotypic harm indexes for IDC cell mitochondria might provide new hallmarks in breast cancer cell biology. No significant financial relationships to disclose.


2005 ◽  
Vol 23 (16_suppl) ◽  
pp. 3210-3210 ◽  
Author(s):  
C. Mundhenke ◽  
M. Weigel ◽  
S. R. Schmidt ◽  
I. Meinhold-Heerlein ◽  
C. Schem ◽  
...  

2017 ◽  
Vol 38 (16) ◽  
pp. NA-NA ◽  
Author(s):  
Sepeedah Soltanian-Zadeh ◽  
Kruthika Kikkeri ◽  
Ayesha N. Shajahan-Haq ◽  
Jeannine Strobl ◽  
Robert Clarke ◽  
...  

Micromachines ◽  
2022 ◽  
Vol 13 (1) ◽  
pp. 93
Author(s):  
Menekse Ermis ◽  
Ezgi Antmen ◽  
Ozgur Kuren ◽  
Utkan Demirci ◽  
Vasif Hasirci

In the recent years, microfabrication technologies have been widely used in cell biology, tissue engineering, and regenerative medicine studies. Today, the implementation of microfabricated devices in cancer research is frequent and advantageous because it enables the study of cancer cells in controlled microenvironments provided by the microchips. Breast cancer is one of the most common cancers in women, and the way breast cancer cells interact with their physical microenvironment is still under investigation. In this study, we developed a transparent cell culture chip (Ch-Pattern) with a micropillar-decorated bottom that makes live imaging and monitoring of the metabolic, proliferative, apoptotic, and morphological behavior of breast cancer cells possible. The reason for the use of micropatterned surfaces is because cancer cells deform and lose their shape and acto-myosin integrity on micropatterned substrates, and this allows the quantification of the changes in morphology and through that identification of the cancerous cells. In the last decade, cancer cells were studied on micropatterned substrates of varying sizes and with a variety of biomaterials. These studies were conducted using conventional cell culture plates carrying patterned films. In the present study, cell culture protocols were conducted in the clear-bottom micropatterned chip. This approach adds significantly to the current knowledge and applications by enabling low-volume and high-throughput processing of the cell behavior, especially the cell–micropattern interactions. In this study, two different breast cancer cell lines, MDA-MB-231 and MCF-7, were used. MDA-MB-231 cells are invasive and metastatic, while MCF-7 cells are not metastatic. The nuclei of these two cell types deformed to distinctly different levels on the micropatterns, had different metabolic and proliferation rates, and their cell cycles were affected. The Ch-Pattern chips developed in this study proved to have significant advantages when used in the biological analysis of live cells and highly beneficial in the study of screening breast cancer cell–substrate interactions in vitro.


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