TGF‑β1‑induced epithelial‑mesenchymal transition increases fatty acid oxidation and OXPHOS activity via the p‑AMPK pathway in breast cancer cells

Liu,Qian‑Qian; Huo,Hong‑Yu; Ao,Song; Liu,Ting; Yang,Li; Fei,Zai‑Yi; Zhang,Zhi‑Qiang; Ding,Lei; Cui,Qing‑Hua; Lin,Jie; Yu,Min; Xiong,Wei

doi:10.3892/or.2020.7661

TGF‑β1‑induced epithelial‑mesenchymal transition increases fatty acid oxidation and OXPHOS activity via the p‑AMPK pathway in breast cancer cells

Authors:
- Qian‑Qian Liu
- Hong‑Yu Huo
- Song Ao
- Ting Liu
- Li Yang
- Zai‑Yi Fei
- Zhi‑Qiang Zhang
- Lei Ding
- Qing‑Hua Cui
- Jie Lin
- Min Yu
- Wei Xiong
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Affiliations: Department of Biochemistry and Genetics, School of Life Sciences, Yunnan University, Kunming, Yunnan 650091, P.R. China, Chifeng Blood Centre, Chifeng, Inner Mongolia, Chifeng 024000, P.R. China, Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Dali University, Dali, Yunnan 671000, P.R. China
Published online on: June 25, 2020 https://doi.org/10.3892/or.2020.7661
Pages: 1206-1215

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Abstract

Breast cancer is the most common malignancy in women, and metastasis is the leading cause of death in breast cancer patients. Previous studies have shown that epithelial‑mesenchymal transition (EMT) is involved in the metastasis of breast cancer, but the metabolic reprogramming and regulation mechanisms involved in the EMT process are still unclear. In the present study, we successfully constructed an EMT cell model induced by transforming growth factor β1 (TGF‑β1) treatment of MCF‑7 cells at different times. The results showed that cell adhesion decreased, cell invasion increased and ATP levels increased in EMT MCF‑7 cells treated with TGF‑β1. Furthermore, the expression of fatty acid synthase (FASN) was decreased, and the expression of key fatty acid β‑oxidation enzymes (CPT1 and CD36) was elevated in treated cells compared to control cells. These results showed that the fatty acid oxidation pathway was enhanced. In addition, the expression of NADH:ubiquinone oxidoreductase subunit B8 (NDUFB8), mitochondrial transcription factor A (TFAM) and cytochrome c oxidase subunit I (COXI) increased, and the mitochondrial DNA copy number and ROS levels were also significantly increased during TGF‑β1‑induced EMT. These results indicated that mitochondrial oxidative phosphorylation (OXPHOS) activity was enhanced during EMT. In addition, we observed that the expression of p‑AMPK was increased and ACC (Acetyl‑CoA Carboxylase) was decreased during TGF‑β1‑induced EMT in MCF‑7 cells. Immunohistochemical analysis of clinical samples revealed high expression of FASN in epithelial cells that had high expression of E‑cadherin, while high expression of CPT‑1 was observed in mesenchymal cells that had high expression of vimentin. Results of the current study showed a metabolic transition in TGF‑β1‑induced EMT in MCF‑7 cells. This transition may regulate fatty acid oxidation and OXPHOS activity in EMT MCF‑7 cells through the p‑AMPK pathway. These data suggest that a metabolic transition that suppresses lipogenesis and favors energy production is an essential component of TGF‑β1‑induced EMT and metastasis in breast cancer. This study thus provides a new strategy for identifying new therapeutic targets for breast cancer.

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1	Anastasiadi Z, Lianos GD, Ignatiadou E, Harissis HV and Mitsis M: Breast cancer in young women: An overview. Updates Surg. 69:313–317. 2017. View Article : Google Scholar : PubMed/NCBI
2	Medeiros B and Allan AL: Molecular mechanisms of breast cancer metastasis to the lung: Clinical and experimental perspectives. Int J Mol Sc. 20:22722019. View Article : Google Scholar
3	Minn AJ, Gupta GP, Siegel PM, Bos PD, Shu W, Giri DD, Viale A, Olshen AB, Gerald WL and Massagué J: Genes that mediate breast cancer metastasis to lung. Nature. 436:518–524. 2005. View Article : Google Scholar : PubMed/NCBI
4	Hu J, Li G, Zhang P, Zhuang X and Hu G: A CD44v⁺ subpopulation of breast cancer stem-like cells with enhanced lung metastasis capacity. Cell Death Dis. 8:e26792017. View Article : Google Scholar : PubMed/NCBI
5	Xu X, Yan Q, Wang Y and Dong X: NTN4 is associated with breast cancer metastasis via regulation of EMT-related biomarkers. Oncol Rep. 37:449–457. 2017. View Article : Google Scholar : PubMed/NCBI
6	Pastushenko I, Brisebarre A, Sifrim A, Fioramonti M, Revenco T, Boumahdi S, Van Keymeulen A, Brown D, Moers V, Lemaire S, et al: Identification of the tumour transition states occurring during EMT. Nature. 556:463–468. 2018. View Article : Google Scholar : PubMed/NCBI
7	Nieto MA, Huang RY, Jackson RA and Thiery JP: EMT: 2016. Cell. 166:21–45. 2016. View Article : Google Scholar : PubMed/NCBI
8	Ungefroren H, Witte D and Lehnert H: The role of small GTPases of the Rho/Rac family in TGF-β-induced EMT and cell motility in cancer. Dev Dyn. 247:451–461. 2018. View Article : Google Scholar : PubMed/NCBI
9	Liu FL, Mo EP, Yang L, Du J, Wang HS, Zhang H, Kurihara H, Xu J and Cai SH: Autophagy is involved in TGF-β1-induced protective mechanisms and formation of cancer-associated fibroblasts phenotype in tumor microenvironment. Oncotarget. 7:4122–4141. 2016. View Article : Google Scholar : PubMed/NCBI
10	Huang R and Zong X: Aberrant cancer metabolism in epithelial-mesenchymal transition and cancer metastasis: Mechanisms in cancer progression. Crit Rev Oncol Hematol. 115:13–22. 2017. View Article : Google Scholar : PubMed/NCBI
11	Chen XL, Lei L, Hong LL and Ling ZQ: Potential role of NDRG2 in reprogramming cancer metabolism and epithelial-to-mesenchymal transition. Histol Histopathol. 33:655–663. 2018.PubMed/NCBI
12	Tan TZ, Miow QH, Miki Y, Noda T, Mori S, Huang RY and Thiery JP: Epithelial-mesenchymal transition spectrum quantification and its efficacy in deciphering survival and drug responses of cancer patients. EMBO Mol Med. 6:1279–1293. 2014. View Article : Google Scholar : PubMed/NCBI
13	Syed V: TGF-β Signaling in Cancer. J Cell Biochem. 117:1279–1287. 2016. View Article : Google Scholar : PubMed/NCBI
14	Kim I and He YY: Targeting the AMP-activated protein kinase for cancer prevention and therapy. Front Oncol. 3:1752013. View Article : Google Scholar : PubMed/NCBI
15	Chiarugi P and Cirri P: Metabolic exchanges within tumor microenvironment. Cancer Lett. 380:272–280. 2016. View Article : Google Scholar : PubMed/NCBI
16	Gouirand V, Guillaumond F and Vasseur S: Influence of the tumor microenvironment on cancer cells metabolic reprogramming. Front Oncol. 8:1172018. View Article : Google Scholar : PubMed/NCBI
17	Cheng S, Wang G, Wang Y, Cai L, Qian K, Ju L, Liu X, Xiao Y and Wang X: Fatty acid oxidation inhibitor etomoxir suppresses tumor progression and induces cell cycle arrest via PPARγ-mediated pathway in bladder cancer. Clin Sci (Lond). 133:1745–1758. 2019. View Article : Google Scholar : PubMed/NCBI
18	Chen RR, Yung MMH, Xuan Y, Zhan S, Leung LL, Liang RR, Leung THY, Yang H, Xu D, Sharma R, et al: Targeting of lipid metabolism with a metabolic inhibitor cocktail eradicates peritoneal metastases in ovarian cancer cells. Commun Biol. 2:2812019. View Article : Google Scholar : PubMed/NCBI
19	Li J, Condello S, Thomes-Pepin J, Ma X, Xia Y, Hurley TD, Matei D and Cheng JX: Lipid desaturation is a metabolic marker and therapeutic target of ovarian cancer stem cells. Cell Stem Cell. 20:303–314.e5. 2017. View Article : Google Scholar : PubMed/NCBI
20	Klein M, Seeger P, Schuricht B, Alper SL and Schwab A: Polarization of Na(+)/H(+) and Cl(−)/HCO (3)(−) exchangers in migrating renal epithelial cells. J Gen Physiol. 115:599–608. 2000. View Article : Google Scholar : PubMed/NCBI
21	Lagana A, Vadnais J, Le PU, Nguyen TN, Laprade R, Nabi IR and Noël J: Regulation of the formation of tumor cell pseudopodia by the Na(+)/H(+) exchanger NHE1. J Cell Sc. 113:3649–3662. 2000.
22	Fan FT, Shen CS, Tao L, Tian C, Liu ZG, Zhu ZJ, Liu YP, Pei CS, Wu HY, Zhang L, et al: PKM2 regulates hepatocellular carcinoma cell epithelial-mesenchymal transition and migration upon EGFR activation. Asian Pac J Cancer Prev. 15:1961–1970. 2014. View Article : Google Scholar : PubMed/NCBI
23	Arseneault R, Chien A, Newington JT, Rappon T, Harris R and Cumming RC: Attenuation of LDHA expression in cancer cells leads to redox-dependent alterations in cytoskeletal structure and cell migration. Cancer Lett. 338:255–266. 2013. View Article : Google Scholar : PubMed/NCBI
24	Liu X, Wang X, Zhang J, Lam EK, Shin VY, Cheng AS, Yu J, Chan FK, Sung JJ and Jin HC: Warburg effect revisited: An epigenetic link between glycolysis and gastric carcinogenesis. Oncogene. 29:442–450. 2010. View Article : Google Scholar : PubMed/NCBI
25	Yang L, Hou Y, Yuan J, Tang S, Zhang H, Zhu Q, Du YE, Zhou M, Wen S, Xu L, et al: Twist promotes reprogramming of glucose metabolism in breast cancer cells through PI3K/AKT and p53 signaling pathways. Oncotarget. 6:25755–25769. 2015. View Article : Google Scholar : PubMed/NCBI
26	Hardie DG, Schaffer BE and Brunet A: AMPK: An energy-sensing pathway with multiple inputs and outputs. Trends Cell Biol. 26:190–201. 2016. View Article : Google Scholar : PubMed/NCBI
27	Carling D: AMPK signalling in health and disease. Current opinion in cell biology. 45:31–37. 2017. View Article : Google Scholar : PubMed/NCBI
28	Wu Y, Sarkissyan M, McGhee E, Lee S and Vadgama JV: Combined inhibition of glycolysis and AMPK induces synergistic breast cancer cell killing. Curr Opin Cell Biol. 151:529–539. 2015.
29	Liu X, Chhipa RR, Nakano I and Dasgupta B: The AMPK inhibitor compound C is a potent AMPK-independent antiglioma agent. Mol Cancer Ther. 13:596–605. 2014. View Article : Google Scholar : PubMed/NCBI
30	Mauro L, Naimo GD, Gelsomino L, Malivindi R, Bruno L, Pellegrino M, Tarallo R, Memoli D, Weisz A, Panno ML and Andò S: Uncoupling effects of estrogen receptor alpha on LKB1/AMPK interaction upon adiponectin exposure in breast cancer. FASEB J. 32:4343–4355. 2018. View Article : Google Scholar : PubMed/NCBI