Insufficient Radiofrequency Ablation Promotes Hepatocellular Carcinoma Metastasis Through N6-Methyladenosine mRNA Methylation-Dependent Mechanism
Tianhong Su
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
These authors contributed equally to the study.Search for more papers by this authorManling Huang
Department of Gastroenterology and Hepatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
These authors contributed equally to the study.Search for more papers by this authorJunbin Liao
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
These authors contributed equally to the study.Search for more papers by this authorShuibin Lin
Center for Translational Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorPeng Yu
Center for Translational Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorJianhua Yang
MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory for Biocontrol, School of Life Sciences, The Fifth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorYuhong Cai
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorShenghua Zhu
Department of Gastroenterology and Hepatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorLixia Xu
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Department of Oncology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorZhenwei Peng
Clinical Trials Unit, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Department of Radiation Oncology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorSui Peng
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Department of Gastroenterology and Hepatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Clinical Trials Unit, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorCorresponding Author
Shuling Chen
Division of Interventional Ultrasound, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO:
Shuling Chen, M.D.
Division of Interventional Ultrasound, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13570485828
or
Ming Kuang, M.D., Ph.D.
Department of Liver Surgery, Precision Medicine Institute, Cancer Center, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13631393958
Search for more papers by this authorCorresponding Author
Ming Kuang
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Cancer Center, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China
ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO:
Shuling Chen, M.D.
Division of Interventional Ultrasound, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13570485828
or
Ming Kuang, M.D., Ph.D.
Department of Liver Surgery, Precision Medicine Institute, Cancer Center, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13631393958
Search for more papers by this authorTianhong Su
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
These authors contributed equally to the study.Search for more papers by this authorManling Huang
Department of Gastroenterology and Hepatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
These authors contributed equally to the study.Search for more papers by this authorJunbin Liao
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
These authors contributed equally to the study.Search for more papers by this authorShuibin Lin
Center for Translational Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorPeng Yu
Center for Translational Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorJianhua Yang
MOE Key Laboratory of Gene Function and Regulation, State Key Laboratory for Biocontrol, School of Life Sciences, The Fifth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorYuhong Cai
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorShenghua Zhu
Department of Gastroenterology and Hepatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorLixia Xu
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Department of Oncology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorZhenwei Peng
Clinical Trials Unit, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Department of Radiation Oncology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorSui Peng
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Department of Gastroenterology and Hepatology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Clinical Trials Unit, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China
Search for more papers by this authorCorresponding Author
Shuling Chen
Division of Interventional Ultrasound, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO:
Shuling Chen, M.D.
Division of Interventional Ultrasound, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13570485828
or
Ming Kuang, M.D., Ph.D.
Department of Liver Surgery, Precision Medicine Institute, Cancer Center, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13631393958
Search for more papers by this authorCorresponding Author
Ming Kuang
Department of Liver Surgery, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China
Cancer Center, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, China
ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO:
Shuling Chen, M.D.
Division of Interventional Ultrasound, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13570485828
or
Ming Kuang, M.D., Ph.D.
Department of Liver Surgery, Precision Medicine Institute, Cancer Center, The First Affiliated Hospital, Sun Yat-sen University
58 Zhong Shan Road 2
Guangzhou, 510080, China
E-mail: [email protected]
Tel.: +86-13631393958
Search for more papers by this authorAbstract
Background and Aims
The dynamic N6-methyladenosine (m6A) mRNA modification is essential for acute stress response and cancer progression. Sublethal heat stress from insufficient radiofrequency ablation (IRFA) has been confirmed to promote HCC progression; however, whether m6A machinery is involved in IRFA-induced HCC recurrence remains open for study.
Approach and Results
Using an IRFA HCC orthotopic mouse model, we detected a higher level of m6A reader YTH N6-methyladenosine RNA binding protein 1-3 (YTHDF1) in the sublethal-heat–exposed transitional zone close to the ablation center than that in the farther area. In addition, we validated the increased m6A modification and elevated YTHDF1 protein level in sublethal-heat–treated HCC cell lines, HCC patient-derived xenograft (PDX) mouse model, and patients’ HCC tissues. Functionally, gain-of-function/loss-of-function assays showed that YTHDF1 promotes HCC cell viability and metastasis. Knockdown of YTHDF1 drastically restrains the tumor metastasis evoked by sublethal heat treatment in tail vein injection lung metastasis and orthotopic HCC mouse models. Mechanistically, we found that sublethal heat treatment increases epidermal factor growth receptor (EGFR) m6A modification in the vicinity of the 5′ untranslated region and promotes its binding with YTHDF1, which enhances the translation of EGFR mRNA. The sublethal-heat–induced up-regulation of EGFR level was further confirmed in the IRFA HCC PDX mouse model and patients’ tissues. Combination of YTHDF1 silencing and EGFR inhibition suppressed the malignancies of HCC cells synergically.
Conclusions
The m6A-YTHDF1-EGFR axis promotes HCC progression after IRFA, supporting the rationale for targeting m6A machinery combined with EGFR inhibitors to suppress HCC metastasis after RFA.
Supporting Information
| Filename | Description |
|---|---|
| hep31766-sup-0001-Supinfo.pdfPDF document, 9.7 MB | Supplementary Material |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
References
- 1Marrero JA, Kulik LM, Sirlin CB, Zhu AX, Finn RS, Abecassis MM, et al. Diagnosis, staging, and management of hepatocellular carcinoma: 2018 practice guidance by the American Association for the Study of Liver Diseases. Hepatology 2018; 68: 723-750.
- 2 European Association for the Study of the Liver, European Organisation for Research and Treatment of Cancer. EASL-EORTC clinical practice guidelines: management of hepatocellular carcinoma. J Hepatol 2012; 56: 908-943.
- 3Heimbach JK, Kulik LM, Finn RS, Sirlin CB, Abecassis MM, Roberts LR, et al. AASLD guidelines for the treatment of hepatocellular carcinoma. Hepatology 2018; 67: 358-380.
- 4Cucchetti A, Piscaglia F, Cescon M, Colecchia A, Ercolani G, Bolondi L, et al. Cost-effectiveness of hepatic resection versus percutaneous radiofrequency ablation for early hepatocellular carcinoma. J Hepatol 2013; 59: 300-307.
- 5Lee DH, Lee JM, Lee JY, Kim SH, Yoon JH, Kim YJ, et al. Radiofrequency ablation of hepatocellular carcinoma as first-line treatment: long-term results and prognostic factors in 162 patients with cirrhosis. Radiology 2014; 270: 900-909.
- 6Lencioni R, Cioni D, Crocetti L, Franchini C, Pina CD, Lera J, et al. Early-stage hepatocellular carcinoma in patients with cirrhosis: long-term results of percutaneous image-guided radiofrequency ablation. Radiology 2005; 234: 961-967.
- 7Rossi S, Ravetta V, Rosa L, Ghittoni G, Viera FT, Garbagnati F, et al. Repeated radiofrequency ablation for management of patients with cirrhosis with small hepatocellular carcinomas: a long-term cohort study. Hepatology 2011; 53: 136-147.
- 8Choi D, Lim HK, Rhim H, Kim YS, Lee WJ, Paik SW, et al. Percutaneous radiofrequency ablation for early-stage hepatocellular carcinoma as a first-line treatment: long-term results and prognostic factors in a large single-institution series. Eur Radiol 2007; 17: 684-692.
- 9Chu KF, Dupuy DDE. Thermal ablation of tumours: biological mechanisms and advances in therapy. Nat Rev Cancer 2014; 14: 199-208.
- 10Su T, Liao J, Dai Z, Xu L, Chen S, Wang Y, et al. Stress-induced phosphoprotein 1 mediates hepatocellular carcinoma metastasis after insufficient radiofrequency ablation. Oncogene 2018; 37: 3514-3527.
- 11Yuan CW, Wang ZC, Liu K, Liu DJ. Incomplete radiofrequency ablation promotes the development of CD133+ cancer stem cells in hepatocellular carcinoma cell line HepG2 via inducing SOX9 expression. Hepatobiliary Pancreat Dis Int 2018; 17: 416-422.
- 12Wang X, Deng Q, Feng K, Chen S, Jiang J, Xia F, et al. Insufficient radiofrequency ablation promotes hepatocellular carcinoma cell progression via autophagy and the CD133 feedback loop. Oncol Rep 2018; 40: 241-251.
- 13Tan LI, Chen S, Wei G, Li Y, Liao J, Jin H, et al. Sublethal heat treatment of hepatocellular carcinoma promotes intrahepatic metastasis and stemness in a VEGFR1-dependent manner. Cancer Lett 2019; 28: 29-40.
10.1016/j.canlet.2019.05.041 Google Scholar
- 14Matilainen O, Quirós PM, Auwerx J. Mitochondria and epigenetics—crosstalk in homeostasis and stress. Trends Cell Biol 2017; 27: 453-463.
- 15Guo S, Jiang X, Wang Y, Chen L, Li H, Li X, et al. The protective role of TET2 in erythroid iron homeostasis against oxidative stress and erythropoiesis. Cell Signal 2017; 38: 106-115.
- 16Jia G, Fu Y, Zhao X, Dai Q, Zheng G, Yang Y, et al. N6-Methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Nat Chem Biol 2011; 7: 885-887.
- 17He C. Grand challenge commentary: RNA epigenetics? Nat Chem Biol 2010; 6: 863-865.
- 18Dominissini D, Moshitch-Moshkovitz S, Schwartz S, Salmon-Divon M, Ungar L, Osenberg S, et al. Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature 2012; 485: 201-206.
- 19Meyer KD, Jaffrey SR. The dynamic epitranscriptome: N6-methyladenosine and gene expression control. Nat Rev Mol Cell Biol 2014; 15: 313-326.
- 20Liu J, Harada BT, He C. Regulation of gene expression by N6-methyladenosine in cancer. Trends Cell Biol 2019; 29: 487-499.
- 21Xiang Y, Laurent B, Hsu CH, Nachtergaele S, Lu Z, Sheng W, et al. RNA m6A methylation regulates the ultraviolet-induced DNA damage response. Nature 2017; 543: 573-576.
- 22Zhou J, Wan J, Gao X, Zhang X, Jaffrey SR, Qian SB. Dynamic m6A mRNA methylation directs translational control of heat shock response. Nature 2015; 526: 591-594.
- 23Zhong LI, Liao D, Zhang M, Zeng C, Li X, Zhang R, et al. YTHDF2 suppresses cell proliferation and growth via destabilizing the EGFR mRNA in hepatocellular carcinoma. Cancer Lett 2019; 442: 252-261.
- 24Ma JZ, Yang F, Zhou CC, Liu F, Yuan JH, Wang F, et al. METTL14 suppresses the metastatic potential of hepatocellular carcinoma by modulating N6-methyladenosine-dependent primary MicroRNA processing. Hepatology 2017; 65: 529-543.
- 25Chen M, Wei L, Law CT, Tsang FC, Shen J, Cheng CH, et al. RNA N6-methyladenosine methyltransferase-like 3 promotes liver cancer progression through YTHDF2-dependent posttranscriptional silencing of SOCS2. Hepatology 2018; 67: 2254-2270.
- 26Fu Y, Dominissini D, Rechavi G, He C. Gene expression regulation mediated through reversible m6A RNA methylation. Nat Rev Genet 2014; 15: 293-306.
- 27Zeng Y, Wang S, Gao S, Soares F, Ahmed M, Guo H, et al. Refined RIP-seq protocol for epitranscriptome analysis with low input materials. PLoS Biol 2018; 16:e2006092.
- 28Dominissini D, Moshitch-Moshkovitz S, Salmon-Divon M, Amariglio N, Rechavi G. Transcriptome-wide mapping of N6-methyladenosine by m6A-seq based on immunocapturing and massively parallel sequencing. Nat Protoc 2013; 8: 176-189.
- 29Wang T, Cui Y, Jin J, Guo J, Wang G, Yin X, et al. Translating mRNAs strongly correlate to proteins in a multivariate manner and their translation ratios are phenotype specific. Nucleic Acids Res 2013; 41: 4743-4754.
- 30Esposito AM, Mateyak M, He D, Lewis M, Sasikumar AN, Hutton J, et al. Eukaryotic polyribosome profile analysis. J Vis Exp 2010; 40:1948.
- 31MacKenzie JA, Payne RM. Preparation of ribosomes loaded with truncated nascent proteins to study ribosome binding to mammalian mitochondria. Mitochondrion 2006; 6: 64-70.
- 32Horn D, Hess J, Freier K, Hoffmann J, Freudlsperger C. Targeting EGFR-PI3K-AKT-mTOR signaling enhances radiosensitivity in head and neck squamous cell carcinoma. Expert Opin Ther Targets 2015; 19: 795-805.
- 33Wang R, Chen L, Chen H, Hu L, Li L, Sun H, et al. MUC15 inhibits dimerization of EGFR and PI3K-AKT signaling and is associated with aggressive hepatocellular carcinomas in patients. Gastroenterology 2013; 145: 1436-1448.e1-e12.
- 34Thompson SM, Jondal DE, Butters KA, Knudsen BE, Anderson JL, Stokes MP, et al. Heat stress induced, ligand-independent MET and EGFR signaling in hepatocellular carcinoma. Int J Hyperthermia 2018; 34: 812-823.
- 35Shi H, Wei J, He C. Where, when, and how: context-dependent functions of RNA methylation writers, readers, and erasers. Mol Cell 2019; 74: 640-650.
- 36Yin Z, Liu XU, Ariosa A, Huang H, Jin M, Karbstein K, et al. Psp2, a novel regulator of autophagy that promotes autophagy-related protein translation. Cell Res 2019; 29: 994-1008.
- 37Schwanhäusser B, Busse D, Li NA, Dittmar G, Schuchhardt J, Wolf J, et al. Global quantification of mammalian gene expression control. Nature 2011; 473: 337-342.
- 38Davalos V, Blanco S, Esteller M. SnapShot: messenger RNA modifications. Cell 2018; 174: 498-498.e1.
- 39Lence T, Paolantoni C, Worpenberg L, Roignant JY. Mechanistic insights into m(6)A RNA enzymes. Biochim Biophys Acta Gene Regul Mech 2019; 1862: 222-229.
- 40Schöller E, Weichmann F, Treiber T, Ringle S, Treiber N, Flatley A, et al. Interactions, localization, and phosphorylation of the m(6)A generating METTL3-METTL14-WTAP complex. RNA 2018; 24: 499-512.
- 41Du Y, Hou G, Zhang H, Dou J, He J, Guo Y, et al. SUMOylation of the m6A-RNA methyltransferase METTL3 modulates its function. Nucleic Acids Res 2018; 46: 5195-5208.
- 42Kim KW, Lee JM, Klotz E, Kim SJ, Kim SH, Kim JY, et al. Safety margin assessment after radiofrequency ablation of the liver using registration of preprocedure and postprocedure CT images. Am J Roentgenol 2011; 196: W565-W572.
Author names in bold designate shared co-first authorship.
