Summary of Study ST002319

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001485. The data can be accessed directly via it's Project DOI: 10.21228/M8S41S This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST002319
Study TitleMass spectroscopy‑based proteomics and metabolomics analysis of triple‑positive breast cancer cells treated with tamoxifen and trastuzumab
Study SummaryHER2-enriched breast cancer with high levels of hormone receptor expression, known as triple positive breast cancer, may represent a new entity with a relatively favourable prognosis against which the combination of chemotherapy, HER-2 inhibition, and endocrine treatment may be considered overtreatment. We explored the effect of the anticancer drugs tamoxifen and trastuzumab, both separately and in combination, on the integrated proteomic and metabolic profile of triple positive breast cancer cells (BT-474). Method We employed ultra-high-performance liquid chromatography-quadrupole time of flight mass spectrometry using a Bruker timsTOF to investigate changes in BT-474 cell line treated with either tamoxifen, trastuzumab or a combination. Differentially abundant metabolites were identified using the Bruker Human Metabolome Database metabolite library and proteins using the Uniprot proteome for Homo sapiens using MetaboScape and MaxQuant, respectively, for identification and quantitation. Results A total of 77 proteins and 85 metabolites were found to significantly differ in abundance in BT-474 treated cells with tamoxifen 5 μM/and or trastuzumab 2.5 μM. Findings suggest that by targeting important cellular signalling pathways which regulate cell growth, apoptosis, proliferation, and chemoresistance, these medicines have a considerable anti-growth effect in BT-474 cells. Pathways enriched for dysregulation include RNA splicing, neutrophil degranulation and activation, cellular redox homeostasis, mitochondrial transmembrane transport, ferroptosis and necroptosis, ABC transporters and central carbon metabolism. Conclusion Our findings in protein and metabolite level research revealed that anti-cancer drug therapy had a significant impact on the key signalling pathways and molecular processes in triple positive BT-474 cell lines.
Institute
University of Sharjah
DepartmentSharjah Institute for Medical Research
LaboratoryBiomarker Discovery Group
Last NameSoares
First NameNelson
AddressM32, SIMR, College of Pharmacy, Health Sciences, University of Sharjah
Emailnsoares@sharjah.ac.ae
Phone065057656
Submit Date2022-10-18
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2022-11-21
Release Version1
Nelson Soares Nelson Soares
https://dx.doi.org/10.21228/M8S41S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP002411
Sampleprep Summary:Sample metabolite extraction A volume of 1 mL of the extraction solvent (methanol+0.1% formic acid) was added to the cell pellets to quench cells. The cells were then vortexed for 2 min to ensure the quantitative extraction of the metabolites and stored on ice for 1 h, during which the samples were vortexed every 15 min. After this, the insoluble cell matrices were collected and transferred to centrifuge tubes, intermittent ultrasonication using the COPLEY sonicator (QSONICA SONICATOR, USA) under 30% amplifier and for 30 s with an ice bath employed throughout the process. Following that, cells debris were then centrifuged (15,000 rpm, 10 min, − 4 °C) and the sample supernatants were collected and transferred to LC vials for drying in the EZ-2 Plus (GeneVac-Ipswich, UK) at 37±1 °C. Dried samples were resuspended with 200 µL (water+0.1% formic acid), and vortexed for 2 min. Finally, the samples were filtered using a hydrophilic Nylon Syringe Filter of 0.45 µm pore size and analyzed by Q-TOF MS
Processing Storage Conditions:Described in summary
Extract Storage:Described in summary
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