PMID- 33926547 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20240401 IS - 2049-3002 (Print) IS - 2049-3002 (Electronic) IS - 2049-3002 (Linking) VI - 9 IP - 1 DP - 2021 Apr 29 TI - TNF-alpha differentially modulates subunit levels of respiratory electron transport complexes of ER/PR +ve/-ve breast cancer cells to regulate mitochondrial complex activity and tumorigenic potential. PG - 19 LID - 10.1186/s40170-021-00254-9 [doi] LID - 19 AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) is an immunostimulatory cytokine that is consistently high in the breast tumor microenvironment (TME); however, its differential role in mitochondrial functions and cell survival in ER/PR +ve and ER/PR -ve breast cancer cells is not well understood. METHODS: In the current study, we investigated TNF-alpha modulated mitochondrial proteome using high-resolution mass spectrometry and identified the differentially expressed proteins in two different breast cancer cell lines, ER/PR positive cell line; luminal, MCF-7 and ER/PR negative cell line; basal-like, MDA-MB-231 and explored its implication in regulating the tumorigenic potential of breast cancer cells. We also compared the activity of mitochondrial complexes, ATP, and ROS levels between MCF-7 and MDA-MB-231 in the presence of TNF-alpha. We used Tumor Immune Estimation Resource (TIMER) webserver to analyze the correlation between TNF-alpha and mitochondrial proteins in basal and luminal breast cancer patients. Kaplan-Meier method was used to analyze the correlation between mitochondrial protein expression and survival of breast cancer patients. RESULTS: The proteome analysis revealed that TNF-alpha differentially altered the level of critical proteins of mitochondrial respiratory chain complexes both in MCF-7 and MDA-MB-231, which correlated with differential assembly and activity of mitochondrial ETC complexes. The inhibition of the glycolytic pathway in the presence of TNF-alpha showed that glycolysis is indispensable for the proliferation and clonogenic ability of MDA-MB-231 cells (ER/PR -ve) as compared to MCF-7 cells (ER/PR +ve). The TIMER database showed a negative correlation between the expressions of TNF-alpha and key regulators of mitochondrial OXPHOS complexes in basal breast vs lobular carcinoma. Conversely, patient survival analysis showed an improved relapse-free survival with increased expression of identified proteins of ETC complexes and survival of the breast cancer patients. CONCLUSION: The evidence presented in our study convincingly demonstrates that TNF-alpha regulates the survival and proliferation of aggressive tumor cells by modulating the levels of critical assembly factors and subunits involved in mitochondrial respiratory chain supercomplexes organization and function. This favors the rewiring of mitochondrial metabolism towards anaplerosis to support the survival and proliferation of breast cancer cells. Collectively, the results strongly suggest that TNF-alpha differentially regulates metabolic adaptation in ER/PR +ve (MCF-7) and ER/PR -ve (MDA-MB-231) cells by modulating the mitochondrial supercomplex assembly and activity. FAU - Shinde, Anjali AU - Shinde A AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. FAU - Jung, Hyeryeon AU - Jung H AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. FAU - Lee, Hayun AU - Lee H AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. FAU - Singh, Kritarth AU - Singh K AD - Department of Cell and Developmental Biology, University College London, Gower Street, London, WC1E 6BT, UK. FAU - Roy, Milton AU - Roy M AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. FAU - Gohel, Dhruv AU - Gohel D AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. FAU - Kim, Han Byeol AU - Kim HB AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. FAU - Mane, Minal AU - Mane M AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. FAU - Vasiyani, Hitesh AU - Vasiyani H AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. FAU - Currim, Fatema AU - Currim F AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. FAU - Seo, Yu Ri AU - Seo YR AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. FAU - Yang, Seojin AU - Yang S AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. FAU - Cho, Ara AU - Cho A AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. FAU - Yi, Eugene C AU - Yi EC AD - Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, 03080, South Korea. euyi@snu.ac.kr. FAU - Singh, Rajesh AU - Singh R AUID- ORCID: 0000-0002-4986-3486 AD - Department of Bio-Chemistry, Faculty of Science, The Maharaja Sayajirao University of Baroda, Sayajigunj, Vadodara, Gujarat, 390002, India. singhraj1975@gmail.com. LA - eng GR - INT/Korea/P-39/Department of Science and Technology, Ministry of Science and Technology/ GR - 2015M3A9B6073835/National Research Foundation of Korea/ GR - NRF-2016R1A5A1010764/National Research Foundation of Korea/ GR - NRF-2017K1A3A1A19071651/National Research Foundation of Korea/ PT - Journal Article DEP - 20210429 PL - England TA - Cancer Metab JT - Cancer & metabolism JID - 101607582 PMC - PMC8082668 OTO - NOTNLM OT - Breast cancer heterogeneity OT - Inflammation OT - Metabolism OT - Mitochondria OT - TNF-alpha COIS- The authors declare that they have no competing interests. EDAT- 2021/05/01 06:00 MHDA- 2021/05/01 06:01 PMCR- 2021/04/29 CRDT- 2021/04/30 05:46 PHST- 2020/09/30 00:00 [received] PHST- 2021/04/01 00:00 [accepted] PHST- 2021/04/30 05:46 [entrez] PHST- 2021/05/01 06:00 [pubmed] PHST- 2021/05/01 06:01 [medline] PHST- 2021/04/29 00:00 [pmc-release] AID - 10.1186/s40170-021-00254-9 [pii] AID - 254 [pii] AID - 10.1186/s40170-021-00254-9 [doi] PST - epublish SO - Cancer Metab. 2021 Apr 29;9(1):19. doi: 10.1186/s40170-021-00254-9.