Circular RNA hsa-circ-0001030 suppresses proliferation and cisplatin tolerance in TSCC via interaction with PKM2

Abstract

Aim: Cisplatin resistance remains a major obstacle to the effective treatment of tongue squamous cell carcinoma (TSCC). This study is dedicated to elucidating the role and mechanism of circular RNA (circRNA) hsa-circ-0001030 in modulating cisplatin sensitivity and metabolic reprogramming in TSCC.

Methods: CircRNA sequencing, quantitative polymerase chain reaction, and RNA fluorescence in situ hybridization were used to test hsa-circ-0001030 expression in TSCC tissues and cell lines. Gain-of-function assays (colony formation, cell counting kit-8, Transwell assay, and xenograft models) were conducted to evaluate proliferation, invasion, and cisplatin response. Mechanistic studies, including RNA pull-down, RNA-binding protein immunoprecipitation, and western blotting, were performed to identify pyruvate kinase M2 (PKM2) as a binding partner of hsa-circ-0001030 and to assess glycolytic activity, glucose uptake, and lactate production.

Results: Hsa-circ-0001030 was markedly downregulated in TSCC and cisplatin-resistant cells. Overexpression of hsa-circ-0001030 suppressed tumor growth, migration, and glycolytic flux, while enhancing cisplatin sensitivity both in vitro and in vivo. Mechanistically, hsa-circ-0001030 directly bound to PKM2 at nucleotides 138-169, inhibited PKM2 enzymatic activity, restraining tetramer formation and increased tyrosine 105 (Tyr105) phosphorylation and thereby blocking PKM2-driven glycolysis. Clinically, low hsa-circ-0001030 expression correlated with advanced tumor-node-metastasis stage, poor differentiation, and unsatisfying prognosis in TSCC patients.

Conclusion: Hsa-circ-0001030 acted as a tumor-suppressive circRNA that might depress PKM2-dependent metabolic reprogramming and cisplatin resistance in TSCC, highlighting its potential as a prognostic biomarker and therapeutic target for overcoming chemoresistance.

Keywords

Tongue squamous cell carcinoma / circRNA / hsa-circ-0001030 / PKM2 / glycolysis / cisplatin resistance / metabolic reprogramming

References

[1] Siegel RL,Jemal A.Cancer statistics, 2024.CA Cancer J Clin2024;74:12-49

[2] Muzaffar J,Kirtane K.Recent advances and future directions in clinical management of head and neck squamous cell carcinoma.Cancers2021;13:338 PMCID:PMC7831487

[3] Ranasinghe R,Zulli A.Cisplatin for cancer therapy and overcoming chemoresistance.Heliyon2022;8:e10608 PMCID:PMC9489975

[4] Faubert B,DeBerardinis RJ.Metabolic reprogramming and cancer progression.Science2020;368:eaaw5473 PMCID:PMC7227780

[5] Heiden MG, Cantley LC, Thompson CB. Understanding the Warburg effect: the metabolic requirements of cell proliferation.Science2009;324:1029-33 PMCID:PMC2849637

[6] Wang RH,Chen YT.Hydrogen sulfide coordinates glucose metabolism switch through destabilizing tetrameric pyruvate kinase M2.Nat Commun2024;15:7463 PMCID:PMC11358145

[7] Upadhyay S,Hassan MI.Exploring the diverse role of pyruvate kinase M2 in cancer: navigating beyond glycolysis and the Warburg effect.Biochim Biophys Acta Rev Cancer2024;1879:189089

[8] Rathore R,Van Tine BA.PHGDH as a mechanism for resistance in metabolically-driven cancers.Cancer Drug Resist2020;3:762-74 PMCID:PMC7840151

[9] Bach DH,Sood AK.Circular RNAs in cancer.Mol Ther Nucleic Acids2019;16:118-29 PMCID:PMC6411617

[10] Conn VM,Conn SJ.Circular RNA in cancer.Nat Rev Cancer2024;24:597-613

[11] Li Q,Zhu D,Jiang R.Circular RNA MAT2B promotes glycolysis and malignancy of hepatocellular carcinoma through the miR-338-3p/PKM2 axis under hypoxic stress.Hepatology2019;70:1298-316

[12] Yang H,Tan Z.Nucleus-targeted Silencer nanoplatform regulating ZEB1-AS1 in head and neck squamous cell carcinoma therapy.Discov Nano2024;19:192 PMCID:PMC11585530

[13] Zhu S,Zhang X.Pyruvate kinase M2 (PKM2) in cancer and cancer therapeutics.Cancer Lett2021;503:240-8

[14] Gu L,Nan X.circCYP24A1 facilitates esophageal squamous cell carcinoma progression through binding PKM2 to regulate NF-κB-induced CCL5 secretion.Mol Cancer2022;21:217 PMCID:PMC9746112

[15] Wong N,Tang D.PKM2, a central point of regulation in cancer metabolism.Int J Cell Biol2013;2013:242513 PMCID:PMC3586519

[16] Pan W,Yin W.The role and clinical applications of exosomes in cancer drug resistance.Cancer Drug Resist2024;7:43 PMCID:PMC11609145

[17] Christofk HR,Wu N,Cantley LC.Pyruvate kinase M2 is a phosphotyrosine-binding protein.Nature2008;452:181-6

[18] Lv L,Zhao D.Acetylation targets the M2 isoform of pyruvate kinase for degradation through chaperone-mediated autophagy and promotes tumor growth.Mol Cell2011;42:719-30 PMCID:PMC4879880

[19] Pan RY,Zhang J.Positive feedback regulation of microglial glucose metabolism by histone H4 lysine 12 lactylation in Alzheimer’s disease.Cell Metab2022;34:634-48.e6

[20] Wang C,Yan S.Interactome analysis reveals that lncRNA HULC promotes aerobic glycolysis through LDHA and PKM2.Nat Commun2020;11:3162 PMCID:PMC7308313

[21] Hua Q,Xu F.Hypoxia-induced lncRNA-AC020978 promotes proliferation and glycolytic metabolism of non-small cell lung cancer by regulating PKM2/HIF-1α axis.Theranostics2020;10:4762-78 PMCID:PMC7163453

[22] Yu Y,Xie F.Tumor-associated macrophage enhances PD-L1-mediated immune escape of bladder cancer through PKM2 dimer-STAT3 complex nuclear translocation.Cancer Lett2024;593:216964

[23] Yang W,Hawke D.PKM2 phosphorylates histone H3 and promotes gene transcription and tumorigenesis.Cell2012;150:685-96 PMCID:PMC3431020

[24] Prakasam G,Bamezai RNK.Posttranslational modifications of pyruvate kinase M2: tweaks that benefit cancer.Front Oncol2018;8:22 PMCID:PMC5808394

[25] Angiari S,Sutton CE.Pharmacological activation of pyruvate kinase M2 inhibits CD4⁺ T cell pathogenicity and suppresses autoimmunity.Cell Metab2020;31:391-405.e8 PMCID:PMC7001035

[26] Li H,Huang Q,Li X.Circular RNA P4HB promotes glycolysis and tumor progression by binding with PKM2 in lung adenocarcinoma.Respir Res2023;24:252 PMCID:PMC10601333

[27] Bian Z,Li M.LncRNA-FEZF1-AS1 promotes tumor proliferation and metastasis in colorectal cancer by regulating PKM2 signaling.Clin Cancer Res2018;24:4808-19

[28] Hitosugi T,Vander Heiden MG.Tyrosine phosphorylation inhibits PKM2 to promote the Warburg effect and tumor growth.Sci Signal2009;2:ra73 PMCID:PMC2812789

[29] Zhang Z,Liu Y,Sun L.PKM2, function and expression and regulation.Cell Biosci2019;9:52 PMCID:PMC6595688

[30] Zheng S,Liu T.Posttranslational modification of pyruvate kinase type M2 (PKM2): novel regulation of its biological roles to be further discovered.J Physiol Biochem2021;77:355-63

[31] Wu B,Lan H,Wang C.The role of PKM2 in cancer progression and its structural and biological basis.J Physiol Biochem2024;80:261-75

[32] Puckett DL,Chowanadisai W.The role of PKM2 in metabolic reprogramming: insights into the regulatory roles of non-coding RNAs.Int J Mol Sci2021;22:1171 PMCID:PMC7865720

[33] Wang Y,Nan Y.PKM2 inhibitor shikonin overcomes the cisplatin resistance in bladder cancer by inducing necroptosis.Int J Biol Sci2018;14:1883-91 PMCID:PMC6231221

[34] Wang D,Xu F.Cisplatin-resistant NSCLC cells induced by hypoxia transmit resistance to sensitive cells through exosomal PKM2.Theranostics2021;11:2860-75 PMCID:PMC7806469