Proteomics Investigation of the Impact of the Enterococcus faecalis Secretome on MCF-7 Tumor Cells

Sep 24, 2026

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Breast cancer is the most prevalent cancer among women, and its tumor microenvironment includes diverse microbiota. Disruption between microbes and host tissues can influence breast cancer development and spread. This study evaluates the impact of Enterococcus faecalis on the luminal A-like MCF-7 cell line using untargeted proteomics to characterize proteomic changes after treatment with bacterial supernatant at 24 and 48 h.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages19

Reading Time48 min

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Introduction

Materials and Methods

Results

Discussion

Conclusions

Article Proteomics Investigation of the Impact of the Enterococcus faecalis Secretome on MCF-7 Tumor Cells Moudi A. Alwehaibi 1,2, Mysoon M. Al-Ansari 1, *, Assim A. Alfadda 2,3, Reem Al-Malki 1, Afshan Masood 2, Anas M. Abdel Rahman 4,5 and Hicham Benabdelkamel 2, * Citation: Alwehaibi, M.A.; Al-Ansari, M.M.; Alfadda, A.A.; Al-Malki, R.; Masood, A.; Abdel Rahman, A.M.; Benabdelkamel, H. Proteomics Investigation of the Impact of the Enterococcus faecalis Secretome on MCF-7 Tumor Cells. Int. J. Mol. Sci. 2023, 24, 14937. [https://doi.org/](https://doi.org/)[ ](https://doi.org/)[10.3390/ijms241914937](10.3390/ijms241914937) Academic Editor: Nicole Hansmeier Received: 3 September 2023 Revised: 2 October 2023 Accepted: 3 October 2023 Published: 5 October 2023 Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ([https://](https://)[ ](https://)[creativecommons.org/licenses/by/](creativecommons.org/licenses/by/)[ ](creativecommons.org/licenses/by/)[4.0/](4.0/)) . 1 Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia; [modhialwehaibi@gmail.com](modhialwehaibi@gmail.com) (M.A.A.); [reem-hm876@hotmail.com](reem-hm876@hotmail.com) (R.A.-M.) 2 Proteomics Resource Unit, Obesity Research Center, College of Medicine, King Saud University, Riyadh 11461, Saudi Arabia; [aalfadda@ksu.edu.sa](aalfadda@ksu.edu.sa) (A.A.A.); [afsmasood@ksu.edu.sa](afsmasood@ksu.edu.sa) (A.M.) 3 Department of Medicine, College of Medicine and King Saud Medical City, King Saud University, Riyadh 11451, Saudi Arabia 4 Metabolomics Section, Department of Clinical Genomics, Center for Genome Medicine, King Faisal Specialist Hospital and Research Centre (KFSHRC), Riyadh 11211, Saudi Arabia; [aabdelrahman46@kfshrc.edu.sa](aabdelrahman46@kfshrc.edu.sa) 5 Department of Biochemistry and Molecular Medicine, College of Medicine, Alfaisal University, Riyadh 11533, Saudi Arabia * [Correspondence: myalansari@ksu.edu.sa](Correspondence: myalansari@ksu.edu.sa) (M.M.A.-A.); [helkamel@ksu.edu.sa](helkamel@ksu.edu.sa) (H.B.) Abstract: Breast cancer is the most prevalent form of cancer among women. The microenvironment of a cancer tumor is surrounded by various cells, including the microbiota. An imbalance between microbes and their host may contribute to the development and spread of breast cancer. Therefore, the objective of this study is to investigate the inﬂuence of Enterococcus faecalis on a breast cancer cell line (MCF-7) to mimic the luminal A subtype of breast cancer, using an untargeted proteomics approach to analyze the proteomic proﬁles of breast cancer cells after their treatment with E. faecalis in order to understand the microbiome and its role in the development of cancer. The breast cancer cell line MCF- 7 was cultured and then treated with a 10% bacterial supernatant at two time points (24 h and 48 h) at 37 􀀎 C in a humidiﬁed incubator with 5% CO2. Proteins were then extracted and separated using two-dimensional difference (2D-DIGE) gel electrophoresis, and the statistically signiﬁcant proteins (p-value < 0.05, fold change > 1.5) were identiﬁed via matrix-assisted laser desorption/ionization– time-of-ﬂight mass spectrometry (MALDI-TOF-MS) . The protein ﬁngerprints showed a differential protein expression pattern in the cells treated with E. faecalis for 24 and 48 h compared with the control. We found 58 statistically signiﬁcant proteins changes in the MCF-7 breast cancer cells affected by E. faecalis. Kilin and transgelin were upregulated after 24 h of treatment and could be used as diagnostic and prognostic markers for breast cancer. In addition, another protein involved in the inhibition of cell proliferation was coiled-coil domain-containing protein 154 . The protein markersidentiﬁed in this study may serve as possible biomark

FAQs

What was the main objective of this study?

To investigate how Enterococcus faecalis affects the breast cancer cell line MCF-7 and to understand the microbiome’s role in breast cancer development.

How were MCF-7 cells treated and analyzed?

MCF-7 cells were cultured and treated with a 10% bacterial supernatant at 24 h and 48 h, then proteins were extracted and analyzed using 2D-DIGE and MALDI-TOF-MS.

Which proteins showed significant changes after treatment?

The protein fingerprints indicated differential expression after 24 and 48 h, with 58 statistically significant protein changes reported. Kilin and transgelin were upregulated after 24 h, and coiled-coil domain-containing protein 154 was linked to inhibition of cell proliferation.

Sep 24, 2026

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