Interference with ENO2 promotes ferroptosis and inhibits glycolysis in clear cell renal cell carcinoma by regulating Hippo‑YAP1 signaling

Li,Hu; Wu,Yanni; Ma,Yong; Liu,Xiaoqiang

doi:10.3892/ol.2024.14576

Interference with ENO2 promotes ferroptosis and inhibits glycolysis in clear cell renal cell carcinoma by regulating Hippo‑YAP1 signaling

Authors:
- Hu Li
- Yanni Wu
- Yong Ma
- Xiaoqiang Liu
View Affiliations

Affiliations: Department of Urology, Tianjin Medical University General Hospital, Tianjin 300052, P.R. China, Department of Medical Technology, Heze Jiazheng Vocational College, Heze, Shandong 274300, P.R. China, Department of Urology, Shanxian Central Hospital, Affiliated Huxi Hospital of Jining Medical University, Heze, Shandong 274300, P.R. China
Published online on: July 19, 2024 https://doi.org/10.3892/ol.2024.14576
Article Number: 443
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Glycolytic enzyme enolase 2 (ENO2) is dysregulated in various cancer types. Nevertheless, the role and underlying mechanism of ENO2 in clear cell renal cell carcinoma (ccRCC) remain unclear. Therefore, the current study investigated the effect and mechanism of ENO2 in ccRCC. ENO2 expression in a ccRCC cell line was assessed using reverse transcription‑quantitative PCR and western blotting. Analysis of glycolysis was performed by estimating the extracellular acidification rate, lactic acid concentration, glucose uptake and the expression of glucose transporter 1, pyruvate kinase muscle isozyme M2 and hexokinase 2. Moreover, ferroptosis was assessed by detecting the level of total iron, lipid peroxide, reactive oxygen species and the expression of ferroptosis‑related protein. In addition, mitochondrial function was assessed using JC‑1 staining and detection kits. The results indicated that ENO2 is expressed at high levels in ccRCC cell lines, and interference with ENO2 expression inhibits glycolysis, promotes ferroptosis and affects mitochondrial function in ccRCC cells. Further investigation demonstrated that interference with ENO2 expression affected ferroptosis levels in ccRCC cells by inhibiting the glycolysis process. Mechanistically, the present results indicated that ENO2 may affect ferroptosis, glycolysis and mitochondrial functions by regulating Hippo‑yes‑associated protein 1 (YAP1) signaling in ccRCC cells. In conclusion, the present study showed that ENO2 affects ferroptosis, glycolysis and mitochondrial functions in ccRCC cells by regulating Hippo‑YAP1 signaling, hence demonstrating its potential as a therapeutic target in ccRCC.

View Figures

View References

1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Motzer RJ, Bacik J and Mazumdar M: Prognostic factors for survival of patients with stage IV renal cell carcinoma: Memorial sloan-kettering cancer center experience. Clin Cancer Res. 10:6302S–6303S. 2004. View Article : Google Scholar : PubMed/NCBI
3	Jonasch E, Gao J and Rathmell WK: Renal cell carcinoma. BMJ. 349:g47972014. View Article : Google Scholar : PubMed/NCBI
4	Pavlova NN and Thompson CB: The emerging hallmarks of cancer metabolism. Cell Metab. 23:27–47. 2016. View Article : Google Scholar : PubMed/NCBI
5	Miranda-Galvis M and Teng Y: Targeting Hypoxia-driven metabolic reprogramming to constrain tumor progression and metastasis. Int J Mol Sci. 21:54872020. View Article : Google Scholar : PubMed/NCBI
6	Zhang Y, Chen M, Liu M, Xu Y and Wu G: Glycolysis-related genes serve as potential prognostic biomarkers in clear cell renal cell carcinoma. Oxid Med Cell Longev. 2021:66998082021.PubMed/NCBI
7	Tang B, Yan R, Zhu J, Cheng S, Kong C, Chen W, Fang S, Wang Y, Yang Y, Qiu R, et al: Integrative analysis of the molecular mechanisms, immunological features and immunotherapy response of ferroptosis regulators across 33 cancer types. Int J Biol Sci. 18:180–198. 2022. View Article : Google Scholar : PubMed/NCBI
8	Torti SV and Torti FM: Iron and cancer: 2020 Vision. Cancer Res. 80:5435–5448. 2020. View Article : Google Scholar : PubMed/NCBI
9	Dixon SJ, Lemberg KM, Lamprecht MR, Skouta R, Zaitsev EM, Gleason CE, Patel DN, Bauer AJ, Cantley AM, Yang WS, et al: Ferroptosis: An iron-dependent form of nonapoptotic cell death. Cell. 149:1060–1072. 2012. View Article : Google Scholar : PubMed/NCBI
10	Hao J, Chen Q, Feng Y, Jiang Q, Sun H, Deng B, Huang X, Guan J, Chen Q, Liu X, et al: Combination treatment with FAAH inhibitors/URB597 and ferroptosis inducers significantly decreases the growth and metastasis of renal cell carcinoma cells via the PI3K-AKT signaling pathway. Cell Death Dis. 14:2472023. View Article : Google Scholar : PubMed/NCBI
11	Reed GH, Poyner RR, Larsen TM, Wedekind JE and Rayment I: Structural and mechanistic studies of enolase. Curr Opin Struct Biol. 6:736–743. 1996. View Article : Google Scholar : PubMed/NCBI
12	Zheng Y, Wu C, Yang J, Zhao Y, Jia H, Xue M, Xu D, Yang F, Fu D, Wang C, et al: Insulin-like growth factor 1-induced enolase 2 deacetylation by HDAC3 promotes metastasis of pancreatic cancer. Signal Transduct Target Ther. 5:532020. View Article : Google Scholar : PubMed/NCBI
13	Marangos PJ, Parma AM and Goodwin FK: Functional properties of neuronal and glial isoenzymes of brain enolase. J Neurochem. 31:727–732. 1978. View Article : Google Scholar : PubMed/NCBI
14	Liu CC, Wang H, Wang WD, Wang L, Liu WJ, Wang JH, Geng QR and Lu Y: ENO2 promotes cell proliferation, glycolysis, and glucocorticoid-resistance in acute lymphoblastic leukemia. Cell Physiol Biochem. 46:1525–1535. 2018. View Article : Google Scholar : PubMed/NCBI
15	Huebbers CU, Adam AC, Preuss SF, Schiffer T, Schilder S, Guntinas-Lichius O, Schmidt M, Klussmann JP and Wiesner RJ: High glucose uptake unexpectedly is accompanied by high levels of the mitochondrial ß-F1-ATPase subunit in head and neck squamous cell carcinoma. Oncotarget. 6:36172–36184. 2015. View Article : Google Scholar : PubMed/NCBI
16	Huang J, Yang M, Liu Z, Li X, Wang J, Fu N, Cao T and Yang X: PPFIA4 promotes colon cancer cell proliferation and migration by enhancing tumor glycolysis. Front Oncol. 11:6532002021. View Article : Google Scholar : PubMed/NCBI
17	Liu C, Liu D, Wang F, Xie J, Liu Y, Wang H, Rong J, Xie J, Wang J, Zeng R, et al: Identification of a glycolysis- and lactate-related gene signature for predicting prognosis, immune microenvironment, and drug candidates in colon adenocarcinoma. Front Cell Dev Biol. 10:9719922022. View Article : Google Scholar : PubMed/NCBI
18	Lang L, Wang F, Ding Z, Zhao X, Loveless R, Xie J, Shay C, Qiu P, Ke Y, Saba NF and Teng Y: Blockade of glutamine-dependent cell survival augments antitumor efficacy of CPI-613 in head and neck cancer. J Exp Clin Cancer Res. 40:3932021. View Article : Google Scholar : PubMed/NCBI
19	Lv C, Yu H, Wang K, Chen C, Tang J, Han F, Mai M, Ye K, Lai M and Zhang H: ENO2 promotes colorectal cancer metastasis by interacting with the LncRNA CYTOR and activating YAP1-induced EMT. Cells. 11:23632022. View Article : Google Scholar : PubMed/NCBI
20	Zheng S, Mo J, Zhang J and Chen Y: HIF-1α inhibits ferroptosis and promotes malignant progression in non-small cell lung cancer by activating the Hippo-YAP signalling pathway. Oncol Lett. 25:902023. View Article : Google Scholar : PubMed/NCBI
21	Liu G, Zhou J, Piao Y, Zhao X, Zuo Y and Ji Z: Hsa_circ_0085576 promotes clear cell renal cell carcinoma tumorigenesis and metastasis through the miR-498/YAP1 axis. Aging (Albany NY). 12:11530–11549. 2020. View Article : Google Scholar : PubMed/NCBI
22	Wang J, Zhao Z, Liu Y, Cao X, Li F, Ran H, Cao Y and Wu C: ‘Mito-Bomb’: A novel mitochondria-targeting nanosystem for ferroptosis-boosted sonodynamic antitumor therapy. Drug Deliv. 29:3111–3122. 2022. View Article : Google Scholar : PubMed/NCBI
23	Zhang M, Zhong H, Cao T, Huang Y, Ji X, Fan GC and Peng T: Gamma-aminobutyrate transaminase protects against lipid overload-triggered cardiac injury in mice. Int J Mol Sci. 23:21822022. View Article : Google Scholar : PubMed/NCBI
24	Huang SX, Partridge MA, Ghandhi SA, Davidson MM, Amundson SA and Hei TK: Mitochondria-derived reactive intermediate species mediate asbestos-induced genotoxicity and oxidative stress-responsive signaling pathways. Environ Health Perspect. 120:840–847. 2012. View Article : Google Scholar : PubMed/NCBI
25	Doherty JR and Cleveland JL: Targeting lactate metabolism for cancer therapeutics. J Clin Invest. 123:3685–3692. 2013. View Article : Google Scholar : PubMed/NCBI
26	Liberti MV and Locasale JW: The Warburg effect: How does it benefit cancer cells? Trends Biochem Sci. 41:211–218. 2016. View Article : Google Scholar : PubMed/NCBI
27	Abbaszadeh Z, Çeşmeli S and Biray Avcı Ç: Crucial players in glycolysis: Cancer progress. Gene. 726:1441582020. View Article : Google Scholar : PubMed/NCBI
28	Guo Y, Liang F, Zhao F and Zhao J: Resibufogenin suppresses tumor growth and Warburg effect through regulating miR-143-3p/HK2 axis in breast cancer. Mol Cell Biochem. 466:103–115. 2020. View Article : Google Scholar : PubMed/NCBI
29	Han S, Yang S, Cai Z, Pan D, Li Z, Huang Z, Zhang P, Zhu H, Lei L and Wang W: Anti-Warburg effect of rosmarinic acid via miR-155 in gastric cancer cells. Drug Des Devel Ther. 9:2695–2703. 2015.PubMed/NCBI
30	Zheng J, Luo J, Zeng H, Guo L and Shao G: ¹²⁵I suppressed the Warburg effect viaregulating miR-338/PFKL axis in hepatocellular carcinoma. Biomed Pharmacother. 119:1094022019. View Article : Google Scholar : PubMed/NCBI
31	Liu Y, Liang T, Qiu X, Ye X, Li Z, Tian B and Yan D: Down-regulation of Nfatc1 suppresses proliferation, migration, invasion, and Warburg effect in prostate cancer cells. Med Sci Monit. 25:1572–1581. 2019. View Article : Google Scholar : PubMed/NCBI
32	Morais M, Dias F, Teixeira AL and Medeiros R: MicroRNAs and altered metabolism of clear cell renal cell carcinoma: Potential role as aerobic glycolysis biomarkers. Biochim Biophys Acta Gen Subj. 1861:2175–2185. 2017. View Article : Google Scholar : PubMed/NCBI
33	Hakimi AA, Reznik E, Lee CH, Creighton CJ, Brannon AR, Luna A, Aksoy BA, Liu EM, Shen R, Lee W, et al: An integrated metabolic atlas of clear cell renal cell carcinoma. Cancer Cell. 29:104–116. 2016. View Article : Google Scholar : PubMed/NCBI
34	Grønningsæter IS, Fredly HK, Gjertsen BT, Hatfield KJ and Bruserud O: Systemic metabolomic profiling of acute myeloid leukemia patients before and during disease-stabilizing treatment based on all-trans retinoic acid, valproic acid, and low-dose chemotherapy. Cells. 8:12292019. View Article : Google Scholar
35	Fang Z, Sun Q, Yang H and Zheng J: SDHB suppresses the tumorigenesis and development of ccRCC by inhibiting glycolysis. Front Oncol. 11:6394082021. View Article : Google Scholar : PubMed/NCBI
36	Kounelakis MG, Zervakis ME, Giakos GC, Postma GJ, Buydens LM and Kotsiakis X: On the relevance of glycolysis process on brain gliomas. IEEE J Biomed Health Inform. 17:128–135. 2013. View Article : Google Scholar : PubMed/NCBI
37	Yeh CS, Wang JY, Chung FY, Lee SC, Huang MY, Kuo CW, Yang MJ and Lin SR: Significance of the glycolytic pathway and glycolysis related-genes in tumorigenesis of human colorectal cancers. Oncol Rep. 19:81–91. 2008.PubMed/NCBI
38	Wang Q, Chen C, Ding Q, Zhao Y, Wang Z, Chen J, Jiang Z, Zhang Y, Xu G, Zhang J, et al: METTL3-mediated m⁶A modification of HDGF mRNA promotes gastric cancer progression and has prognostic significance. Gut. 69:1193–1205. 2020. View Article : Google Scholar : PubMed/NCBI
39	Sanders E and Diehl S: Analysis and interpretation of transcriptomic data obtained from extended Warburg effect genes in patients with clear cell renal cell carcinoma. Oncoscience. 2:151–186. 2015. View Article : Google Scholar : PubMed/NCBI
40	Teng PN, Hood BL, Sun M, Dhir R and Conrads TP: Differential proteomic analysis of renal cell carcinoma tissue interstitial fluid. J Proteome Res. 10:1333–1342. 2011. View Article : Google Scholar : PubMed/NCBI
41	Rasmuson T, Grankvist K and Ljungberg B: Serum gamma-enolase and prognosis of patients with renal cell carcinoma. Cancer. 72:1324–1328. 1993. View Article : Google Scholar : PubMed/NCBI
42	Takashi M, Sakata T and Kato K: Use of serum gamma-enolase and aldolase A in combination as markers for renal cell carcinoma. Jpn J Cancer Res. 84:304–309. 1993. View Article : Google Scholar : PubMed/NCBI
43	Qiu Y, Cao Y, Cao W, Jia Y and Lu N: The application of ferroptosis in diseases. Pharmacol Res. 159:1049192020. View Article : Google Scholar : PubMed/NCBI
44	Li Y, Feng D, Wang Z, Zhao Y, Sun R, Tian D, Liu D, Zhang F, Ning S, Yao J and Tian X: Ischemia-induced ACSL4 activation contributes to ferroptosis-mediated tissue injury in intestinal ischemia/reperfusion. Cell Death Differ. 26:2284–2299. 2019. View Article : Google Scholar : PubMed/NCBI
45	Zou Y, Henry WS, Ricq EL, Graham ET, Phadnis VV, Maretich P, Paradkar S, Boehnke N, Deik AA, Reinhardt F, et al: Plasticity of ether lipids promotes ferroptosis susceptibility and evasion. Nature. 585:603–608. 2020. View Article : Google Scholar : PubMed/NCBI
46	Wang J, Yin X, He W, Xue W, Zhang J and Huang Y: SUV39H1 deficiency suppresses clear cell renal cell carcinoma growth by inducing ferroptosis. Acta Pharm Sin B. 11:406–419. 2021. View Article : Google Scholar : PubMed/NCBI
47	Kumfu S, Chattipakorn S, Fucharoen S and Chattipakorn N: Mitochondrial calcium uniporter blocker prevents cardiac mitochondrial dysfunction induced by iron overload in thalassemic mice. Biometals. 25:1167–1175. 2012. View Article : Google Scholar : PubMed/NCBI
48	Fang X, Wang H, Han D, Xie E, Yang X, Wei J, Gu S, Gao F, Zhu N, Yin X, et al: Ferroptosis as a target for protection against cardiomyopathy. Proc Natl Acad Sci USA. 116:2672–2680. 2019. View Article : Google Scholar : PubMed/NCBI
49	She H, Tan L, Du Y, Zhou Y, Guo N, Zhang J, Du Y, Wang Y, Wu Z, Ma C, et al: VDAC2 malonylation participates in sepsis-induced myocardial dysfunction via mitochondrial-related ferroptosis. Int J Biol Sci. 19:3143–3158. 2023. View Article : Google Scholar : PubMed/NCBI
50	Anastasiou D, Yu Y, Israelsen WJ, Jiang JK, Boxer MB, Hong BS, Tempel W, Dimov S, Shen M, Jha A, et al: Pyruvate kinase M2 activators promote tetramer formation and suppress tumorigenesis. Nat Chem Biol. 8:839–847. 2012. View Article : Google Scholar : PubMed/NCBI
51	Yang L, Xie M, Yang M, Yu Y, Zhu S, Hou W, Kang R, Lotze MT, Billiar TR, Wang H, et al: PKM2 regulates the Warburg effect and promotes HMGB1 release in sepsis. Nat Commun. 5:44362014. View Article : Google Scholar : PubMed/NCBI
52	Rao J, Wang H, Ni M, Wang Z, Wang Z, Wei S, Liu M, Wang P, Qiu J, Zhang L, et al: FSTL1 promotes liver fibrosis by reprogramming macrophage function through modulating the intracellular function of PKM2. Gut. 71:2539–2550. 2022. View Article : Google Scholar : PubMed/NCBI
53	Zhou X, Xie F, Wang L, Zhang L, Zhang S, Fang M and Zhou F: The function and clinical application of extracellular vesicles in innate immune regulation. Cell Mol Immunol. 17:323–334. 2020. View Article : Google Scholar : PubMed/NCBI