N6-methyladenosine-mediated upregulation of LNCAROD confers radioresistance in esophageal squamous cell carcinoma through stabilizing PARP1

Xiaobo Shi , Xiaozhi Zhang , Xinran Huang , Ruijuan Zhang , Shupei Pan , Shan Huang , Yuchen Wang , Yue Ke , Wei Guo , Xiaoxiao Liu , Yu Hao , You Li , Xu Zhao , Yuchen Sun , Jing Li , Hongbing Ma , Xixi Zhao

Clinical and Translational Medicine ›› 2024, Vol. 14 ›› Issue (10) : e70039

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Clinical and Translational Medicine ›› 2024, Vol. 14 ›› Issue (10) : e70039 DOI: 10.1002/ctm2.70039
RESEARCH ARTICLE

N6-methyladenosine-mediated upregulation of LNCAROD confers radioresistance in esophageal squamous cell carcinoma through stabilizing PARP1

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Abstract

Background: Radiotherapy is a primary therapeutic modality for esophageal squamous cell carcinoma (ESCC), but its effectiveness is still restricted due to the resistance of cancer cells to radiation. Long non-coding RNAs (lncRNAs) and N6-methyladenosine (m6A) have been shown to play significant roles in tumour radioresistance. However, the precise manifestation and role of m6A-modified lncRNAs in ESCC radioresistance remain unclear.

Methods: Bioinformatics analysis was conducted to identify m6A-modified lncRNAs implicated in the radioresistance of ESCC. A series of functional experiments were performed to investigate the function of LNCAROD in ESCC. Methylated RNA immunoprecipitation, chromatin isolation by RNA purification-mass spectrometry, RNA immunoprecipitation, and co-immunoprecipitation experiments were performed to explore the mechanism of m6A-mediated upregulation of LNCAROD expression and the downstream mechanism enhancing the radioresistance of ESCC. The efficacy of LNCAROD in vivo was assessed using murine xenograft models.

Results: Herein, we identified LNCAROD as a novel METTL3-mediated lncRNA that enhanced radioresistance in ESCC cells and was post-transcriptionally stabilised by YTHDC1. Moreover, we confirmed that LNCAROD prevented ubiquitin-proteasome degradation of PARP1 protein by facilitating PARP1-NPM1 interaction, thereby contributing to homologous recombination-mediated DNA double-strand breaks repair and enhancing the radiation resistance of ESCC cells. Silencing LNCAROD in a nude mouse model of ESCC in vivo resulted in slower tumour growth and increased radiosensitivity.

Conclusion: Our findings enhance the understanding of m6A-modified lncRNA-driven machinery in ESCC radioresistance and underscore the significance of LNCAROD in this context, thereby contributing to the development of a potential therapeutic target for ESCC patients.

Keywords

esophageal squamous cell carcinoma / LNCAROD / N6-methyladenosine / PARP1 / radioresistance

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Xiaobo Shi, Xiaozhi Zhang, Xinran Huang, Ruijuan Zhang, Shupei Pan, Shan Huang, Yuchen Wang, Yue Ke, Wei Guo, Xiaoxiao Liu, Yu Hao, You Li, Xu Zhao, Yuchen Sun, Jing Li, Hongbing Ma, Xixi Zhao. N6-methyladenosine-mediated upregulation of LNCAROD confers radioresistance in esophageal squamous cell carcinoma through stabilizing PARP1. Clinical and Translational Medicine, 2024, 14(10): e70039 DOI:10.1002/ctm2.70039

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References

[1]

Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021; 71(3): 209-249.

[2]

Chen R, Zheng R, Zhang S, et al. Patterns and trends in esophageal cancer incidence and mortality in China: an analysis based on cancer registry data. J Nat Cancer Center. 2023; 3(1): 21-27.

[3]

He Y, Liang D, Du L, et al. Clinical characteristics and survival of 5283 esophageal cancer patients: a multicenter study from eighteen hospitals across six regions in China. Cancer Commun (Lond, Engl). 2020; 40(10): 531-544.

[4]

Lagergren J, Smyth E, Cunningham D, et al. Oesophageal cancer. Lancet (London, England). 2017; 390(10110): 2383-2396.

[5]

Thrift AP. Global burden and epidemiology of Barrett oesophagus and oesophageal cancer. Nat Rev Gastroenterol Hepatol. 2021; 18(6): 432-443.

[6]

Jiang X, Liu B, Nie Z, et al. The role of m6A modification in the biological functions and diseases. Signal Transduct Targeted Ther. 2021; 6(1): 74.

[7]

Yan X, Pei K, Guan Z, et al. AI-empowered integrative structural characterization of m(6)A methyltransferase complex. Cell Res. 2022; 32(12): 1124-1127.

[8]

Liu J, Yue Y, Han D, et al. A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Nat Chem Biol. 2014; 10(2): 93-95.

[9]

Frye M, Harada BT, Behm M, et al. RNA modifications modulate gene expression during development. Science. 2018; 361(6409): 1346-1349.

[10]

Zheng G, Dahl JA, Niu Y, et al. ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. Mol Cell. 2013; 49(1): 18-29.

[11]

Jia G, Fu Y, Zhao X, et al. N6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Nat Chem Biol. 2011; 7(12): 885-887.

[12]

Patil DP, Pickering BF, Jaffrey SR. Reading m(6)A in the transcriptome: m(6)A-binding proteins. Trends Cell Biol. 2018; 28(2): 113-127.

[13]

Meyer KD, Jaffrey SR. Rethinking m(6)A readers, writers, and erasers. Annu Rev Cell Dev Biol. 2017; 33: 319-342.

[14]

Visvanathan A, Patil V, Arora A, et al. Essential role of METTL3-mediated m(6)A modification in glioma stem-like cells maintenance and radioresistance. Oncogene. 2018; 37(4): 522-533.

[15]

Zhang C, Chen L, Peng D, et al. METTL3 and N6-methyladenosine promote homologous recombination-mediated repair of DSBs by modulating DNA-RNA hybrid accumulation. Mol Cell. 2020; 79(3): 425-442.

[16]

Wang Y, Zhang L, Sun XL, et al. NRP1 contributes to stemness and potentiates radioresistance via WTAP-mediated m6A methylation of Bcl-2 mRNA in breast cancer. Apoptosis. 2023; 28(1-2): 233-246.

[17]

Shi X, Liu X, Pan S, et al. A novel autophagy-related long non-coding RNA signature to predict prognosis and therapeutic response in esophageal squamous cell carcinoma. Int J Gen Med. 2021; 14: 8325-8339.

[18]

Shi X, Liu X, Huang S, et al. miR-4443 promotes radiation resistance of esophageal squamous cell carcinoma via targeting PTPRJ. J Transl Med. 2022; 20(1): 626.

[19]

Gu Z, Zhu W, Wang W, et al. Anlotinib inhibits tumor angiogenesis and promotes the anticancer effect of radiotherapy on esophageal cancer through inhibiting EphA2. J Oncol. 2022; 2022: 5632744.

[20]

Zhou Y, Zeng P, Li YH, et al. SRAMP: prediction of mammalian N6-methyladenosine (m6A) sites based on sequence-derived features. Nucleic Acids Res. 2016; 44(10): e91.

[21]

Zhao X, Ma Y, Li J, et al. The AEG-1-USP10-PARP1 axis confers radioresistance in esophageal squamous cell carcinoma via facilitating homologous recombination-dependent DNA damage repair. Cancer Lett. 2023; 577: 216440.

[22]

Wang W, Shao F, Yang X, et al. METTL3 promotes tumour development by decreasing APC expression mediated by APC mRNA N(6)-methyladenosine-dependent YTHDF binding. Nat Commun. 2021; 12(1): 3803.

[23]

Han H, Yang C, Zhang S, et al. METTL3-mediated m(6)A mRNA modification promotes esophageal cancer initiation and progression via Notch signaling pathway. Mol Ther Nucl Acids. 2021; 26: 333-346.

[24]

Chen L, Zhang C, Ma W, et al. METTL3-mediated m6A modification stabilizes TERRA and maintains telomere stability. Nucleic Acids Res. 2022; 50(20): 11619-11634.

[25]

Mah LJ, El-Osta A, Karagiannis TC. gammaH2AX: a sensitive molecular marker of DNA damage and repair. Leukemia. 2010; 24(4): 679-686.

[26]

Takano S, Shibamoto Y, Wang Z, et al. Optimal timing of a γH2AX analysis to predict cellular lethal damage in cultured tumor cell lines after exposure to diagnostic and therapeutic radiation doses. J Radiat Res (Tokyo). 2023; 64(2): 317-327.

[27]

Zong C, Zhu T, He J, et al. PARP mediated DNA damage response, genomic stability and immune responses. Int J Cancer. 2022; 150(11): 1745-1759.

[28]

Sekhar KR, Freeman ML. Nucleophosmin plays a role in repairing DNA damage and is a target for cancer treatment. Cancer Res. 2023; 83(10): 1573-1580.

[29]

Goldstein M, Kastan MB. The DNA damage response: implications for tumor responses to radiation and chemotherapy. Annu Rev Med. 2015; 66: 129-143.

[30]

Groelly FJ, Fawkes M, Dagg RA, et al. Targeting DNA damage response pathways in cancer. Nat Rev Cancer. 2023; 23(2): 78-94.

[31]

Ray Chaudhuri A, Nussenzweig A. The multifaceted roles of PARP1 in DNA repair and chromatin remodelling. Nat Rev Mol Cell Biol. 2017; 18(10): 610-621.

[32]

Xu Y, Dong B, Zhu W, et al. A Phase III multicenter randomized clinical trial of 60 Gy versus 50 Gy radiation dose in concurrent chemoradiotherapy for inoperable esophageal squamous cell carcinoma. Clin Cancer. 2022; 28(9): 1792-1799.

[33]

An L, Li M, Jia Q. Mechanisms of radiotherapy resistance and radiosensitization strategies for esophageal squamous cell carcinoma. Mol Cancer. 2023; 22(1): 140.

[34]

Sun Y, Wang J, Ma Y, et al. Radiation induces NORAD expression to promote ESCC radiotherapy resistance via EEPD1/ATR/Chk1 signalling and by inhibiting pri-miR-199a1 processing and the exosomal transfer of miR-199a-5p. J Exp Clin Cancer Res: CR. 2021; 40(1): 306.

[35]

Zhou Y, Shao Y, Hu W, et al. A novel long noncoding RNA SP100-AS1 induces radioresistance of colorectal cancer via sponging miR-622 and stabilizing ATG3. Cell Death Differ. 2023; 30(1): 111-124.

[36]

Ban Y, Tan P, Cai J, et al. LNCAROD is stabilized by m6A methylation and promotes cancer progression via forming a ternary complex with HSPA1A and YBX1 in head and neck squamous cell carcinoma. Mol Oncol. 2020; 14(6): 1282-1296.

[37]

Ansari S, Nikpour P. LNCAROD promotes the proliferation and migration of gastric cancer: a bioinformatics analysis and experimental validation. Funct Integr Genomics. 2023; 23(1): 34.

[38]

Roundtree IA, Evans ME, Pan T, et al. Dynamic RNA modifications in gene expression regulation. Cell. 2017; 169(7): 1187-1200.

[39]

Liu WW, Zhang ZY, Wang F, et al. Emerging roles of m6A RNA modification in cancer therapeutic resistance. Exp Hematol Oncol. 2023; 12(1): 21.

[40]

Xu X, Zhang P, Huang Y, et al. METTL3-mediated m6A mRNA contributes to the resistance of carbon-ion radiotherapy in non-small-cell lung cancer. Cancer Sci. 2023; 114(1): 105-114.

[41]

Alemasova EE, Lavrik OI. Poly(ADP-ribosyl)ation by PARP1: reaction mechanism and regulatory proteins. Nucleic Acids Res. 2019; 47(8): 3811-3827.

[42]

Ziv O, Zeisel A, Mirlas-Neisberg N, et al. Identification of novel DNA-damage tolerance genes reveals regulation of translesion DNA synthesis by nucleophosmin. Nat Commun. 2014; 5: 5437.

[43]

Xu R, Yu S, Zhu D, et al. hCINAP regulates the DNA-damage response and mediates the resistance of acute myelocytic leukemia cells to therapy. Nat Commun. 2019; 10(1): 3812.

[44]

Qin G, Wang X, Ye S, et al. NPM1 upregulates the transcription of PD-L1 and suppresses T cell activity in triple-negative breast cancer. Nat Commun. 2020; 11(1): 1669.

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2024 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.

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