scholarly journals Real-time scratch assay reveals mechanisms of early calcium signaling in breast cancer cells in response to wounding

Oncotarget ◽  
2018 ◽  
Vol 9 (38) ◽  
pp. 25008-25024 ◽  
Author(s):  
Stephen J.P. Pratt ◽  
Erick O. Hernández-Ochoa ◽  
Rachel M. Lee ◽  
Eleanor C. Ory ◽  
James S. Lyons ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Calcium Signaling ◽  
Cancer Cells ◽  
Real Time ◽  
Breast Cancer Cells ◽  
Scratch Assay

scholarly journals Secretion of Mutant DNA and mRNA by the Exosomes of Breast Cancer Cells

Molecules ◽  
2021 ◽  
Vol 26 (9) ◽  
pp. 2499
Author(s):  
Olga E. Andreeva ◽  
Yuri Y. Shchegolev ◽  
Alexander M. Scherbakov ◽  
Ekaterina I. Mikhaevich ◽  
Danila V. Sorokin ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Breast Cancer Cells ◽  
Sanger Sequencing ◽  
Mcf7 Cells ◽  
Rna Transcripts ◽  
Dna And Rna ◽  
Allele Specific

Exosomes are the small vesicles that are secreted by different types of normal and tumour cells and can incorporate and transfer their cargo to the recipient cells. The main goal of the present work was to study the tumour exosomes’ ability to accumulate the parent mutant DNA or RNA transcripts with their following transfer to the surrounding cells. The experiments were performed on the MCF7 breast cancer cells that are characterized by the unique coding mutation in the PIK3CA gene. Using two independent methods, Sanger sequencing and allele-specific real-time PCR, we revealed the presence of the fragments of the mutant DNA and RNA transcripts in the exosomes secreted by the MCF7 cells. Furthermore, we demonstrated the MCF7 exosomes’ ability to incorporate into the heterologous MDA-MB-231 breast cancer cells supporting the possible transferring of the exosomal cargo into the recipient cells. Sanger sequencing of the DNA from MDA-MB-231 cells (originally bearing a wild type of PIK3CA) treated with MCF7 exosomes showed no detectable amount of mutant DNA or RNA; however, using allele-specific real-time PCR, we revealed a minor signal from amplification of a mutant allele, showing a slight increase of mutant DNA in the exosome-treated MDA-MB-231 cells. The results demonstrate the exosome-mediated secretion of the fragments of mutant DNA and mRNA by the cancer cells and the exosomes’ ability to transfer their cargo into the heterologous cells.

scholarly journals Negative regulation of CDC42 expression and cell cycle progression by miR-29a in breast cancer

Open Medicine ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. 78-82 ◽  
Author(s):  
Mingliang Zhang ◽  
Wei Guo ◽  
Jun Qian ◽  
Benzhong Wang
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Cancer Cells ◽  
Real Time ◽  
Cell Lines ◽  
Breast Cancer Cells ◽  
Cell Cycle Progression ◽  
Negative Regulation ◽  
Luciferase Assay ◽  
Cycle Progression

AbstractObjectiveThe inhibitory role of microRNA-29a (miR-29a) has been assessed in breast cancer cells. Herein, we analyze the underlying mechanisms of its role in cell cycle progression in breast cancer cells.MethodsWe applied real-time polymerase chain reaction (PCR) to detect the expression of miR-29 in breast cancer cell lines. Then one of the cell lines, MDA-MB-453, was transfected with mimics of miR-29a. The cell cycle was analyzed by fluorescence-activated cell sorting after staining the cells with propidium iodide. Real-time PCR, luciferase assay and western blot were used together to verify the regulation of the predicted target, cell division cycle 42 (CDC42) by miR-29a.ResultsMiR-29s were decreased in our selected mammary cell lines, among which miR-29a was the dominant isoform. Overexpression of miR-29a caused cell cycle arrest at the G0/G1 phase. We further found that miR-29a could target the expression of CDC42, which is a small GTPase associated with cell cycle progression.ConclusionWe suggest that miR-29a exerts its tumor suppressor role in breast cancer cells partially by arresting the cell cycle through negative regulation of CDC42.

Real-time imaging of cancer cell chemotaxis in paper-based scaffolds

The Analyst ◽  
2016 ◽  
Vol 141 (2) ◽  
pp. 661-668 ◽  
Author(s):  
Rachael M. Kenney ◽  
Matthew W. Boyce ◽  
Andrew S. Truong ◽  
C. Robert Bagnell ◽  
Matthew R. Lockett
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Real Time ◽  
Cancer Cell ◽  
Breast Cancer Cells ◽  
Invasion Assay ◽  
Real Time Imaging ◽  
Cell Chemotaxis

An easy to assemble paper-based invasion assay to study chemotaxis of breast cancer cells in gradients of oxygen in real-time.

scholarly journals The role of vulpinic acid as a natural compound in the regulation of breast cancer-associated miRNAs

2021 ◽  
Vol 54 (1) ◽  
Author(s):  
Demet Cansaran-Duman ◽  
Sevcan Yangın ◽  
Betül Çolak
Keyword(s):  
Breast Cancer ◽  
Cell Proliferation ◽  
Cancer Treatment ◽  
Cancer Cells ◽  
Real Time ◽  
Microarray Analysis ◽  
Mirna Expression ◽  
Breast Cancer Cells ◽  
Mcf 7

Abstract Background Breast cancer is the most frequently diagnosed cancer, and no effective treatment solution has yet been found. The number of studies based on the research of novel natural compounds in the treatment of breast cancer has been increasing in recent years. The anticancer properties of natural compounds are related to the regulation of microRNA (miRNA) expression. Therefore, changing the profile of miRNAs with the use of natural products is very important in cancer treatment. However, the role of vulpinic acid and related miRNAs in breast cancer progression remains unknown. Vulpinic acid, methyl (as2E)-2-(3-hydroxy-5-oxo-4-phenylfuran-2-ylidene)-2 phenylacetate, is a natural product extracted from the lichen species and shows an anticancer effect on different cancer cells. Methods This study examines the effects of vulpinic acid on the miRNA levels of breast cancer (MCF-7) cells and its relationship with cell proliferation and apoptosis levels. The antiproliferative effect of vulpinic acid was screened against MCF-7 breast cancer cells and MCF-12A breast epithelial cells using the xCELLigence real-time cell analysis system. We analyzed the altered miRNA expression profile in MCF-7 breast cancer cells versus MCF-12A cells following their response to vulpinic acid through microarray analysis. The microarray analysis results were confirmed through quantitative real-time PCR and bioinformatics analysis. Results The results of the miRNA array and bioinformatic analyses demonstrated that 12 miRNAs were specifically responsive to vulpinic acid in MCF-7 breast cancer cells. This is the first study to reveal that vulpinic acid inhibits the expression of 12 miRNAs and suppresses breast cancer cell proliferation. The study also revealed that vulpinic acid may downregulate the expression of 12 miRNAs by repressing the FOXO-3 gene. The miRNA targets were mainly found to play a role in the apoptosis, cell cycle and MAPK pathways. Moreover, Bcl-2, Bax, procaspase-3 and procaspase-9 protein levels were assessed by western blot analysis for validation of apoptosis at the protein level. Conclusion This study revealed the molecular mechanisms of vulpinic acid on breast cancer and showed that vulpinic acid regulates apoptosis signaling pathways by decreasing the expression of miRNAs. The miRNA expression patterns illuminate the underlying effect of vulpinic acid in breast cancer treatment. Graphical Abstract

Bioenergetic differences between MCF-7 and T47D breast cancer cells and their regulation by oestradiol and tamoxifen

2014 ◽  
Vol 465 (1) ◽  
pp. 49-61 ◽  
Author(s):  
Brandie N. Radde ◽  
Margarita M. Ivanova ◽  
Huy Xuan Mai ◽  
Joshua K. Salabei ◽  
Bradford G. Hill ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Real Time ◽  
Oestrogen Receptor ◽  
Breast Cancer Cells ◽  
Flux Analysis ◽  
Intact Cells ◽  
Extracellular Flux ◽  
Oestrogen Receptor Α ◽  
Mcf 7

Oestrogen receptor α-expressing breast cancer cells show differences in basal bioenergetics profiles and bioenergetics responses to serum depletion, oestradiol and tamoxifen as measured in real time by extracellular flux analysis in intact cells.

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