Prolyl hydroxylase 2 inhibits glycolytic activity in colorectal cancer via the NF‑κB signaling pathway

Xiang,Lisha; Wei,Hao; Ye,Wentao; Wu,Shuang; Xie,Ganfeng

doi:10.3892/ijo.2023.5590

Prolyl hydroxylase 2 inhibits glycolytic activity in colorectal cancer via the NF‑κB signaling pathway

Authors:
- Lisha Xiang
- Hao Wei
- Wentao Ye
- Shuang Wu
- Ganfeng Xie
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Affiliations: Division of Thoracic Tumor Multimodality Treatment and Department of Medical Oncology, Cancer Center, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, West China Medical School, West China Medical Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Department of Oncology and Southwest Cancer Center, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing 400038, P.R. China
Published online on: November 15, 2023 https://doi.org/10.3892/ijo.2023.5590
Article Number: 2
Copyright: © Xiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A variety of malignancies preferentially meet energy demands through the glycolytic pathway. Hypoxia‑induced cancer cell adaptations are essential for tumor development. However, in cancerous glycolysis, the functional importance and underlying molecular mechanism of prolyl hydroxylase domain protein 2 (PHD2) have not been fully elucidated. Gain‑ and loss‑of‑function assays were conducted to evaluate PHD2 functions in colon cancer cells. Glucose uptake, lactate production and intracellular adenosine‑5'‑triphosphate/adenosine diphosphate ratio were measured to determine glycolytic activities. Protein and gene expression levels were measured by western blot analysis and reverse transcription‑quantitative PCR, respectively. The human colon cancer xenograft model was used to confirm the role of PHD2 in tumor progression in vivo. Functionally, the data demonstrated that PHD2 knockdown leads to increased glycolysis, while PHD2 overexpression resulted in suppressed glycolysis in colorectal cancer cells. In addition, the glycolytic activity was enhanced without PHD2 and normalized after PHD2 reconstitution. PHD2 was shown to inhibit colorectal tumor growth, suppress cancer cell proliferation and improve tumor‑bearing mice survival in vivo. Mechanically, it was found that PHD2 inhibits the expression of critical glycolytic enzymes (glucose transporter 1, hexokinase 2 and phosphoinositide‑dependent protein kinase 1). In addition, PHD2 inhibited Ikkβ‑mediated NF‑κB activation in a hypoxia‑inducible factor‑1α‑independent manner. In conclusion, the data demonstrated that PHD2/Ikkβ/NF‑κB signaling has critical roles in regulating glycolysis and suggests that PHD2 potentially suppresses colorectal cancer.

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1	Dang CV: Cancer metabolism: The known, unknowns. Biochim Biophys Acta Rev Cancer. 1870:12018.
2	Martinez-Reyes I and Chandel NS: Cancer metabolism: Looking forward. Nat Rev Cancer. 21:669–680. 2021.
3	Trefts E and Shaw RJ: AMPK: Restoring metabolic homeostasis over space and time. Mol Cell. 81:3677–3690. 2021.
4	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011.
5	Zecchin A, Kalucka J, Dubois C and Carmeliet P: How endothelial cells adapt their metabolism to form vessels in tumors. Front Immunol. 8:17502017.
6	Losman JA, Koivunen P and Kaelin WG Jr: 2-Oxoglutarate-dependent dioxygenases in cancer. Nat Rev Cancer. 20:710–726. 2020.
7	Liao C and Zhang Q: Understanding the oxygen-sensing pathway and its therapeutic implications in diseases. Am J Pathol. 190:1584–1595. 2020.
8	Semenza GL: The Genomics and genetics of oxygen homeostasis. Annu Rev Genomics Hum Genet. 21:183–204. 2020.
9	Koivunen P and Kietzmann T: Hypoxia-inducible factor prolyl 4-hydroxylases and metabolism. Trends Mol Med. 24:1021–1035. 2018.
10	Wong BW, Kuchnio A, Bruning U and Carmeliet P: Emerging novel functions of the oxygen-sensing prolyl hydroxylase domain enzymes. Trends Biochem Sci. 38:3–11. 2013.
11	Maxwell PH and Eckardt KU: HIF prolyl hydroxylase inhibitors for the treatment of renal anaemia and beyond. Nat Rev Nephrol. 12:157–168. 2016.
12	Singh L, Aldosary S, Saeedan AS, Ansari MN and Kaithwas G: Prolyl hydroxylase 2: A promising target to inhibit hypoxia-induced cellular metabolism in cancer cells. Drug Discov Today. 23:1873–1882. 2018.
13	Kato H, Inoue T, Asanoma K, Nishimura C, Matsuda T and Wake N: Induction of human endometrial cancer cell senescence through modulation of HIF-1alpha activity by EGLN1. Int J Cancer. 118:1144–1153. 2006.
14	Kaelin WG Jr: The VHL tumor suppressor gene: Insights into oxygen sensing and cancer. Trans Am Clin Climatol Assoc. 128:298–307. 2017.
15	Chen F, Chen J, Yang L, Liu J, Zhang X, Zhang Y, Tu Q, Yin D, Lin D, Wong PP, et al: Extracellular vesicle-packaged HIF-1α-stabilizing lncRNA from tumour-associated macrophages regulates aerobic glycolysis of breast cancer cells. Nat Cell Biol. 21:498–510. 2019.
16	Sadiku P, Willson JA, Dickinson RS, Murphy F, Harris AJ, Lewis A, Sammut D, Mirchandani AS, Ryan E, Watts ER, et al: Prolyl hydroxylase 2 inactivation enhances glycogen storage and promotes excessive neutrophilic responses. J Clin Invest. 127:3407–3420. 2017.
17	Guentsch A, Beneke A, Swain L, Farhat K, Nagarajan S, Wielockx B, Raithatha K, Dudek J, Rehling P, Zieseniss A, et al: PHD2 is a regulator for glycolytic reprogramming in macrophages. Mol Cell Biol. 37:e00236–16. 2016.
18	Xiang L, Gilkes DM, Hu H, Takano N, Luo W, Lu H, Bullen JW, Samanta D, Liang H and Semenza GL: Hypoxia-inducible factor 1 mediates TAZ expression and nuclear localization to induce the breast cancer stem cell phenotype. Oncotarget. 5:12509–12527. 2014.
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.
20	Farooq SM, Hou Y, Li H, O'Meara M, Wang Y, Li C and Wang JM: Disruption of GPR35 exacerbates dextran sulfate sodium-induced colitis in mice. Dig Dis Sci. 63:2910–2922. 2018.
21	Wang S, Wu Y, Hou Y, Guan X, Castelvetere MP, Oblak JJ, Banerjee S, Filtz TM, Sarkar FH, Chen X, et al: CXCR2 macromolecular complex in pancreatic cancer: A potential therapeutic target in tumor growth. Transl Oncol. 6:216–225. 2013.
22	Ruifrok AC and Johnston DA: Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol. 23:291–299. 2001.
23	Xie G, Zheng L, Ou J, Huang H, He J, Li J, Pan F and Liang H: Low expression of prolyl hydroxylase 2 is associated with tumor grade and poor prognosis in patients with colorectal cancer. Exp Biol Med (Maywood). 237:860–866. 2012.
24	Zhou L, Wang Y, Zhou M, Zhang Y, Wang P, Li X, Yang J, Wang H and Ding Z: HOXA9 inhibits HIF-1α-mediated glycolysis through interacting with CRIP2 to repress cutaneous squamous cell carcinoma development. Nat Commun. 9:14802018.
25	Dupuy F, Tabariès S, Andrzejewski S, Dong Z, Blagih J, Annis MG, Omeroglu A, Gao D, Leung S, Amir E, et al: PDK1-dependent metabolic reprogramming dictates metastatic potential in breast cancer. Cell Metab. 22:577–589. 2015.
26	Denko NC: Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer. 8:705–713. 2008.
27	Jaakkola P, Mole DR, Tian YM, Wilson MI, Gielbert J, Gaskell SJ, von Kriegsheim A, Hebestreit HF, Mukherji M, Schofield CJ, et al: Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science. 292:468–472. 2001.
28	Vidimar V, Licona C, Cerón-Camacho R, Guerin E, Coliat P, Venkatasamy A, Ali M, Guenot D, Le Lagadec R, Jung AC, et al: A redox ruthenium compound directly targets PHD2 and inhibits the HIF1 pathway to reduce tumor angiogenesis independently of p53. Cancer Lett. 440-441:145–155. 2019.
29	Meneses AM and Wielockx B: PHD2: From hypoxia regulation to disease progression. Hypoxia (Auckl). 4:53–67. 2016.
30	Cummins EP, Berra E, Comerford KM, Ginouves A, Fitzgerald KT, Seeballuck F, Godson C, Nielsen JE, Moynagh P, Pouyssegur J and Taylor CT: Prolyl hydroxylase-1 negatively regulates IkappaB kinase-beta, giving insight into hypoxia-induced NFkappaB activity. Proc Natl Acad Sci USA. 103:18154–18159. 2006.
31	Chan DA, Kawahara TLA, Sutphin PD, Chang HY, Chi JT and Giaccia AJ: Tumor vasculature is regulated by PHD2-mediated angiogenesis and bone marrow-derived cell recruitment. Cancer Cell. 15:527–538. 2009.
32	Hoesel B and Schmid JA: The complexity of NF-κB signaling in inflammation and cancer. Mol Cancer. 12:862013.
33	Birbach A, Gold P, Binder BR, Hofer E, de Martin R and Schmid JA: Signaling molecules of the NF-kappa B pathway shuttle constitutively between cytoplasm and nucleus. J Biol Chem. 277:10842–10851. 2002.
34	Rückert F, Grützmann R and Pilarsky C: Feedback within the inter-cellular communication and tumorigenesis in carcinomas. PLoS One. 7:e367192012.
35	Stine ZE, Walton ZE, Altman BJ, Hsieh AL and Dang CV: MYC, metabolism, and cancer. Cancer Discov. 5:1024–1039. 2015.
36	Obacz J, Pastorekova S, Vojtesek B and Hrstka R: Cross-talk between HIF and p53 as mediators of molecular responses to physiological and genotoxic stresses. Mol Cancer. 12:932013.
37	Johnson RF and Perkins ND: Nuclear factor-κB, p53, and mitochondria: Regulation of cellular metabolism and the Warburg effect. Trends Biochem Sci. 37:317–324. 2012.
38	Wilde L, Roche M, Domingo-Vidal M, Tanson K, Philp N, Curry J and Martinez-Outschoorn U: Metabolic coupling and the reverse Warburg effect in cancer: Implications for novel biomarker and anticancer agent development. Semin Oncol. 44:198–203. 2017.
39	Wang X, Liu R, Qu X, Yu H, Chu H, Zhang Y, Zhu W, Wu X, Gao H, Tao B, et al: α-Ketoglutarate-activated NF-κB signaling promotes compensatory glucose uptake and brain tumor development. Mol Cell. 76:148–162.e7. 2019.
40	Ishak Gabra MB, Yang Y, Lowman XH, Reid MA, Tran TQ and Kong M: IKKβ activates p53 to promote cancer cell adaptation to glutamine deprivation. Oncogenesis. 7:932018.
41	Ruf M, Pitzen T, Meyer C and Drumm J: Atlantoaxial rotatory dislocation: Delayed diagnose will result in more invasive treatment options. J Neurol Surg A Cent Eur Neurosurg. 82:1–8. 2021.
42	Taniguchi K and Karin M: NF-κB, inflammation, immunity and cancer: Coming of age. Nat Rev Immunol. 18:309–324. 2018.
43	Gaete D, Rodriguez D, Watts D, Sormendi S, Chavakis T and Wielockx B: HIF-prolyl hydroxylase domain proteins (PHDs) in cancer-potential targets for anti-tumor therapy? Cancers (Basel). 13:9882021.
44	Chen T, Zhou Q, Tang H, Bozkanat M, Yuan JX, Raj JU and Zhou G: miR-17/20 controls prolyl hydroxylase 2 (PHD2)/hypoxia-inducible factor 1 (HIF1) to regulate pulmonary artery smooth muscle cell proliferation. J Am Heart Assoc. 5:e0045102016.
45	Lee G, Won HS, Lee YM, Choi JW, Oh TI, Jang JH, Choi DK, Lim BO, Kim YJ, Park JW, et al: Oxidative dimerization of PHD2 is responsible for its inactivation and contributes to metabolic reprogramming via HIF-1alpha activation. Sci Rep. 6:189282016.
46	Li J, Yuan W, Jiang S, Ye W, Yang H, Shapiro IM and Risbud MV: Prolyl-4-hydroxylase domain protein 2 controls NF-κB/p65 transactivation and enhances the catabolic effects of inflammatory cytokines on cells of the nucleus pulposus. J Biol Chem. 290:7195–7207. 2015.
47	Wang L, Niu Z, Wang X, Li Z, Liu Y, Luo F and Yan X: PHD2 exerts anti-cancer and anti-inflammatory effects in colon cancer xenografts mice via attenuating NF-κB activity. Life Sci. 242:1171672020.
48	Lee DC, Sohn HA, Park ZY, Oh S, Kang YK, Lee KM, Kang M, Jang YJ, Yang SJ and Hong YK: A lactate-induced response to hypoxia. Cell. 161:595–609. 2015.
49	Romero-Ruiz A, Bautista L, Navarro V, Heras-Garvín A, March-Díaz R, Castellano A, Gómez-Díaz R, Castro MJ, Berra E, López-Barneo J and Pascual A: Prolyl hydroxylase-dependent modulation of eukaryotic elongation factor 2 activity and protein translation under acute hypoxia. J Biol Chem. 287:9651–9658. 2012.
50	Wang X, Xie J and Proud CG: Eukaryotic elongation factor 2 kinase (eEF2K) in cancer. Cancers. 9:1622017.
51	Xue J, Li X, Jiao S, Wei Y, Wu G and Fang J: Prolyl hydroxylase-3 is down-regulated in colorectal cancer cells and inhibits IKKbeta independent of hydroxylase activity. Gastroenterology. 138:606–615. 2010.
52	Lind DS, Hochwald SN, Malaty J, Rekkas S, Hebig P, Mishra G, Moldawer LL, Copeland EM III and Mackay S: Nuclear factor-kappa B is upregulated in colorectal cancer. Surgery. 130:363–369. 2001.
53	Wang Z, Sheng C, Kan G, Yao C, Geng R and Chen S: RNAi screening identifies that TEX10 promotes the proliferation of colorectal cancer cells by increasing NF-κB activation. Adv Sci (Weinh). 7:20005932020.
54	Goka ET, Chaturvedi P, Lopez DTM, Garza A and Lippman ME: RAC1b overexpression confers resistance to chemotherapy treatment in colorectal cancer. Mol Cancer Ther. 18:957–968. 2019.
55	Gyamfi J, Kim J and Choi J: Cancer as a metabolic disorder. Int J Mol Sci. 23:11552022.
56	Ge T, Gu X, Jia R, Ge S, Chai P, Zhuang A and Fan X: Crosstalk between metabolic reprogramming and epigenetics in cancer: Updates on mechanisms and therapeutic opportunities. Cancer Commun (Lond). 42:1049–1082. 2022.