scholarly journals Evaluation of MDA-MB-468 Cell Culture Media Analysis in Predicting Triple-Negative Breast Cancer Patient Sera Metabolic Profiles

Metabolites ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 173
Author(s):  
Wojciech Wojtowicz ◽  
Anna Wróbel ◽  
Karolina Pyziak ◽  
Radosław Tarkowski ◽  
Alicja Balcerzak ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Culture Media ◽  
Media Analysis ◽  
Serum Samples ◽  
Cell Culture Media ◽  
Time Courses ◽  
And Control

Triple-negative breast cancer (TNBC) is characterized by limited survival, poor prognosis, and high recurrence. Understanding the metabolic adaptations of TNBC could help reveal improved treatment regiments. Here we performed a comprehensive 1H NMR metabolic characterization of the MDA-MB-468 cell line, a commonly used model of TNBC, followed by an analysis of serum samples obtained from TNBC patients and healthy controls. MDA-MB-468 cells were cultured, and changes in the metabolic composition of the medium were monitored for 72 h. Based on time courses, metabolites were categorized as being consumed, being produced, or showing a mixed behavior. When comparing TNBC and control samples (HC), and by using multivariate and univariate analyses, we identified nine metabolites with differing profiles). The serum of TNBC patients was characterized by higher levels of glucose, glutamine, citrate, and acetoacetate and by lower levels of lactate, alanine, tyrosine, glutamate, and acetone. A comparative analysis between MDA-MB-468 cell culture media and TNBC patients’ serum identified a potential systemic response to the carcinogenesis-associated processes, highlighting that MDA-MB-468 cells footprint does not reflect metabolic changes observed in studied TNBC serum fingerprint.

scholarly journals Sulforaphane Modulates Cytokine Levels in Cell Culture Media of Triple-Negative Breast Cancer Cells Grown with Macrophages

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 337-337
Author(s):  
Courtney Merrick ◽  
Lauren Housley
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Breast Cancer Cells ◽  
Triple Negative ◽  
Culture Media ◽  
California State University ◽  
State University ◽  
Cell Culture Media ◽  
Cytokine Levels ◽  
The Media

Abstract Objectives Triple negative breast cancer (TNBC) makes up approximately 10–20% of all breast cancer cases and is more common in younger women and in Hispanic and African American populations. It is particularly difficult to treat, exhibiting high-metastasis rates, poor prognosis, and limited treatment options. Mortality from TNBC is largely due to the tumor cells high invasive capacity and rapid progression to metastasis. Evidence suggests that macrophages in the breast tumor microenvironment release cytokines that increase tumor cell proliferation, invasion and metastasis. Sulforaphane (SFN) is a broccoli phytochemical that has been identified to slow the progression of breast cancer as well as alter cytokine secretion from macrophages and breast cancer cells grown in single culture. SFN effects on cytokine secretion in the breast tumor microenvironment remain unclear. This study is investigating the effect of SFN on cytokine levels in cell culture media of TNBC cells grown with and without macrophages. Our hypothesis is that cytokine levels differ in media from cocultured cells versus singly cultured cells, and SFN treatment further alters cytokine levels in media. Methods In this study, TNBC cells (MDA-MB-231) were grown in transwell plates with and without macrophages (THP-1 cells differentiated with phorbol-myristate acetate). Cell cultures (n = 3) were treated with either 15 µM SFN, DMSO (vehicle-control), or a non-treatment control. We evaluated the levels of 44 individual cytokines in cell culture media at 24 and 48 hours after treatment using a multi-plex (BioPlex) assay. Control groups included single-cultured MDA-MB-231 and differentiated THP-1 cells. Results Preliminary analyses revealed that cytokine levels differed in the media of single versus cocultured cells and among treatment groups after 24 and 48 hours of treatment. Conclusions The profile of cytokines in the media of TNBC cells grown with macrophages was influenced by SFN treatment. This information may help establish mechanisms underlying SFN effects on TNBC behavior and identify new treatment strategies. Funding Sources California State University Program for Education and Research in Biotechnology, California State University-Chico.

scholarly journals Evaluation of 6-mercaptopurine in a cell culture model of adaptable triple-negative breast cancer with metastatic potential

Oncotarget ◽  
2019 ◽  
Vol 10 (38) ◽  
pp. 3681-3693
Author(s):  
Balraj Singh ◽  
Vanessa N. Sarli ◽  
Hannah E. Kinne ◽  
Anna Shamsnia ◽  
Anthony Lucci
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Metastatic Potential ◽  
Cell Culture Model ◽  
Culture Model ◽  
A Cell

scholarly journals Degradation of silicon photonic biosensors in cell culture media: analysis and prevention

2017 ◽  
Vol 8 (6) ◽  
pp. 2924
Author(s):  
Graham J. Triggs ◽  
Gareth J. O. Evans ◽  
Thomas F. Krauss
Keyword(s):  
Cell Culture ◽  
Culture Media ◽  
Media Analysis ◽  
Cell Culture Media ◽  
Silicon Photonic ◽  
Photonic Biosensors

scholarly journals A DNA methylation-based liquid biopsy for triple-negative breast cancer

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Katrina Cristall ◽  
Francois-Clement Bidard ◽  
Jean-Yves Pierga ◽  
Michael J. Rauh ◽  
Tatiana Popova ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Liquid Biopsy ◽  
Blood Test ◽  
Triple Negative ◽  
Superior Performance ◽  
Serum Samples ◽  
Clonal Hematopoiesis ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

AbstractHere, we present a next-generation sequencing (NGS) methylation-based blood test called methylation DETEction of Circulating Tumour DNA (mDETECT) designed for the optimal detection and monitoring of metastatic triple-negative breast cancer (TNBC). Based on a highly multiplexed targeted sequencing approach, this assay incorporates features that offer superior performance and included 53 amplicons from 47 regions. Analysis of a previously characterised cohort of women with metastatic TNBC with limited quantities of plasma (<2 ml) produced an AUC of 0.92 for detection of a tumour with a sensitivity of 76% for a specificity of 100%. mDETECTTNBC was quantitative and showed superior performance to an NGS TP53 mutation-based test carried out on the same patients and to the conventional CA15-3 biomarker. mDETECT also functioned well in serum samples from metastatic TNBC patients where it produced an AUC of 0.97 for detection of a tumour with a sensitivity of 93% for a specificity of 100%. An assay for BRCA1 promoter methylation was also incorporated into the mDETECT assay and functioned well but its clinical significance is currently unclear. Clonal Hematopoiesis of Indeterminate Potential was investigated as a source of background in control subjects but was not seen to be significant, though a link to adiposity may be relevant. The mDETECTTNBC assay is a liquid biopsy able to quantitatively detect all TNBC cancers and has the potential to improve the management of patients with this disease.

scholarly journals circUSP42 Is Downregulated in Triple-Negative Breast Cancer and Associated With Poor Prognosis

2020 ◽  
Vol 19 ◽  
pp. 153303382095082
Author(s):  
Jinling Yu ◽  
Weida Shen ◽  
Jinping Xu ◽  
Bo Gong ◽  
Beimin Gao ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Clinical Stage ◽  
Rank Test ◽  
Circular Rnas ◽  
Sequencing Data ◽  
Control Groups ◽  
Qrt Pcr ◽  
And Control

We previously showed that microRNA-182 (miR-182) might promote cell proliferation and migration in triple-negative breast cancer (TNBC). This study aimed to investigate circular RNAs (circRNAs) that interact with miR-182 and play important roles in TNBC. Thirty patients with TNBC were enrolled. One pair of tumor and adjacent tissue samples (control) were submitted for circRNA sequencing to establish the expression profile of circRNAs. Concomitantly, circRNAs aberrantly expressed between TNBC and control groups were identified, and these differentially expressed circRNAs (DEcircRNAs) were subjected to Gene Ontology and KEGG pathway enrichment analyses, as well as prediction of interactions with miRNAs. The expression levels of 5 circRNAs interacting with miR-182 were validated using qRT-PCR. Associations between the expression of circUSP42 and clinicopathological features and prognosis were evaluated. A total of 825 upregulated and 1127 downregulated DEcircRNAs were identified between tumor and control groups. Upregulated DEcircRNAs were significantly involved in proteoglycans in cancer, and endocytosis. Downregulated DEcircRNAs were involved in the pathway of resistance to EGFR tyrosine kinase inhibitors. Prediction of circRNA-miRNA interactions showed that hsa_circ_0002032, chr6:131973682-132047340+, hsa_circ_0005982, hsa_circ_0007823 (circUSP42), and hsa_circ_0001777 might act as miRNA sponges for miR-182. qRT-PCR showed consistent results with circRNA sequencing data ( P < 0.05). Downregulation of circUSP42 was significantly associated with lymph node metastasis ( P = 0.005) and advanced clinical stage ( P = 0.032). Furthermore, Kaplan-Meier plots showed that low expression of circUSP42 was closely associated with poor outcome (log-rank test, P < 0.001). Our data suggested that dysregulation of circUSP42 might contribute to the development and progression of TNBC.

scholarly journals The New Synthetic Serum-Free Medium OptiPASS Promotes High Proliferation and Drug Efficacy Prediction on Spheroids from MDA-MB-231 and SUM1315 Triple-Negative Breast Cancer Cell Lines

2019 ◽  
Vol 8 (3) ◽  
pp. 397 ◽  
Author(s):  
Clémence Dubois ◽  
Pierre Daumar ◽  
Corinne Aubel ◽  
Jean Gauthier ◽  
Bernard Vidalinc ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Cancer Cell Lines ◽  
Breast Cancer Cell Lines ◽  
Serum Free

Triple-negative breast cancers are particularly aggressive. In vitro cultures are one of the major pathways for developing anticancer strategies. The effectiveness and reproducibility of the drug screenings depend largely on the homogeneity of culture media. In order to optimize the predictive responses of triple-negative breast cancer 3D cell culture models, these works were focused on the development of SUM1315 and MDA-MB-231 cell lines in OptiPASS medium, a new serum-free formulation (BIOPASS). In monolayer cell culture, OptiPASS medium was more suitable for MDA-MB-231 than SUM1315 cell line but maintained cell phenotype and allowed sufficient proliferation. For spheroids produced in OptiPASS, the size monitoring showed a 1.3 and 1.5-fold increase for MDA-MB-231 and SUM1315 cell lines, respectively and viability/mortality profiles were maintained. Spheroids drug sensitivity thresholds were also improved allowing quicker high throughput drug screenings. These results showed the suitability of OptiPASS for 2D and 3D cell cultures of these two triple-negative breast cancer cell lines, with reproducibility of spheroid formation superior to 98%. This opens the way to the common use of this synthetic medium in future preclinical breast cancer research studies.

Targeted metabolomics reveals dynamic portrayal of amino acids and derivatives in triple-negative breast cancer cells and culture media

2021 ◽  
Author(s):  
Fang Kou ◽  
Bangjie Zhu ◽  
Wenbin Zhou ◽  
Chunming Lv ◽  
Yu Cheng ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Amino Acids ◽  
Cancer Cells ◽  
Triple Negative Breast Cancer ◽  
Breast Cancer Cells ◽  
Triple Negative ◽  
Culture Media ◽  
Targeted Metabolomics ◽  
Amino Acids And Derivatives

Targeted metabolomics reveals dynamic portrayal of amino acids and derivatives in triple-negative breast cancer cells and culture media.

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