The epitranscriptome m6A writer METTL3 promotes chemo- and radioresistance in pancreatic cancer cells

  • Authors:
    • Kosuke Taketo
    • Masamitsu Konno
    • Ayumu Asai
    • Jun Koseki
    • Masayasu Toratani
    • Taroh Satoh
    • Yuichiro Doki
    • Masaki Mori
    • Hideshi Ishii
    • Kazuhiko Ogawa
  • View Affiliations

  • Published online on: December 7, 2017     https://doi.org/10.3892/ijo.2017.4219
  • Pages: 621-629
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

N6-methyladenosine (m6A) is the most abundant epitranscriptome modification in mammalian mRNA. Recent years have seen substantial progress in m6A epitranscriptomics, indicating its crucial roles in the initiation and progression of cancer through regulation of RNA stabilities, mRNA splicing, microRNA processing and mRNA translation. However, by what means m6A is dynamically regulated or written by enzymatic components represented by methyltransferase-like 3 (METTL3) and how m6A is significant for each of the numerous genes remain unclear. We focused on METTL3 in pancreatic cancer, the prognosis of which is not satisfactory despite the development of multidisciplinary therapies. We established METTL3-knockdown pancreatic cancer cell line using short hairpin RNA. Although morphologic and proliferative changes were unaffected, METTL3-depleted cells showed higher sensitivity to anticancer reagents such as gemcitabine, 5-fluorouracil, cisplatin and irradiation. Our data suggest that METTL3 is a potent target for enhancing therapeutic efficacy in patients with pancreatic cancer. In addition, we performed cDNA expression analysis followed by gene ontology and protein-protein interaction analysis using the Database for Annotation, Visualization, and Integrated Discovery and Search Tool for the Retrieval of Interacting Genes/Proteins databases, respectively. The results demonstrate that METTL3 was associated with mitogen-activated protein kinase cascades, ubiquitin-dependent process and RNA splicing and regulation of cellular process, suggesting functional roles and targets of METTL3.

References

1 

Waddington CH: The epigenotype. Endeavour. 1:18–20. 1942.

2 

Cantara WA, Crain PF, Rozenski J, McCloskey JA, Harris KA, Zhang X, Vendeix FA, Fabris D and Agris PF: The RNA Modification Database, RNAMDB: 2011 update. Nucleic Acids Res. 39(Database): D195–D201. 2011. View Article : Google Scholar :

3 

Ping XL, Sun BF, Wang L, Xiao W, Yang X, Wang WJ, Adhikari S, Shi Y, Lv Y, Chen YS, et al: Mammalian WTAP is a regulatory subunit of the RNA N6-methyladenosine methyltrans-ferase. Cell Res. 24:177–189. 2014. View Article : Google Scholar : PubMed/NCBI

4 

Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng X, et al: A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. Nat Chem Biol. 10:93–95. 2014. View Article : Google Scholar :

5 

Perry RP and Kelley DE: Existence of methylated messenger RNA in mouse L cells. Cell. 1:37–42. 1974. View Article : Google Scholar

6 

Desrosiers R, Friderici K and Rottman F: Identification of methylated nucleosides in messenger RNA from Novikoff hepatoma cells. Proc Natl Acad Sci USA. 71:3971–3975. 1974. View Article : Google Scholar : PubMed/NCBI

7 

Dominissini D, Moshitch-Moshkovitz S, Schwartz S, Salmon-Divon M, Ungar L, Osenberg S, Cesarkas K, Jacob-Hirsch J, Amariglio N, Kupiec M, et al: Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature. 485:201–206. 2012. View Article : Google Scholar : PubMed/NCBI

8 

Meyer KD, Saletore Y, Zumbo P, Elemento O, Mason CE and Jaffrey SR: Comprehensive analysis of mRNA methylation reveals enrichment in 3′ UTRs and near stop codons. Cell. 149:1635–1646. 2012. View Article : Google Scholar : PubMed/NCBI

9 

Wang X, Lu Z, Gomez A, Hon GC, Yue Y, Han D, Fu Y, Parisien M, Dai Q, Jia G, et al: N6-methyladenosine-dependent regulation of messenger RNA stability. Nature. 505:117–120. 2014. View Article : Google Scholar

10 

Lin S, Choe J, Du P, Triboulet R and Gregory RI: The m6A Methyltransferase METTL3 promotes translation in human cancer cells. Mol Cell. 62:335–345. 2016. View Article : Google Scholar : PubMed/NCBI

11 

Batista PJ, Molinie B, Wang J, Qu K, Zhang J, Li L, Bouley DM, Lujan E, Haddad B, Daneshvar K, et al: m6A RNA modification controls cell fate transition in mammalian embryonic stem cells. Cell Stem Cell. 15:707–719. 2014. View Article : Google Scholar : PubMed/NCBI

12 

Jia G, Fu Y, Zhao X, Dai Q, Zheng G, Yang Y, Yi C, Lindahl T, Pan T, Yang YG, et al: N6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO. Nat Chem Biol. 7:885–887. 2011. View Article : Google Scholar : PubMed/NCBI

13 

Zheng G, Dahl JA, Niu Y, Fedorcsak P, Huang CM, Li CJ, Vågbø CB, Shi Y, Wang WL, Song SH, et al: ALKBH5 is a mammalian RNA demethylase that impacts RNA metabolism and mouse fertility. Mol Cell. 49:18–29. 2013. View Article : Google Scholar :

14 

Liu N, Dai Q, Zheng G, He C, Parisien M and Pan T: N6-methyladenosine-dependent RNA structural switches regulate RNA-protein interactions. Nature. 518:560–564. 2015. View Article : Google Scholar : PubMed/NCBI

15 

Fustin JM, Doi M, Yamaguchi Y, Hida H, Nishimura S, Yoshida M, Isagawa T, Morioka MS, Kakeya H, Manabe I, et al: RNA-methylation-dependent RNA processing controls the speed of the circadian clock. Cell. 155:793–806. 2013. View Article : Google Scholar : PubMed/NCBI

16 

Du H, Zhao Y, He J, Zhang Y, Xi H, Liu M, Ma J and Wu L: YTHDF2 destabilizes m6A-containing RNA through direct recruitment of the CCR4-NOT deadenylase complex. Nat Commun. 7:126262016. View Article : Google Scholar

17 

Wang Y, Li Y, Toth JI, Petroski MD, Zhang Z and Zhao JC: N6-methyladenosine modification destabilizes developmental regulators in embryonic stem cells. Nat Cell Biol. 16:191–198. 2014. View Article : Google Scholar : PubMed/NCBI

18 

Meyer KD, Patil DP, Zhou J, Zinoviev A, Skabkin MA, Elemento O, Pestova TV, Qian SB and Jaffrey SR: 5′ UTR m6A promotes cap-independent translation. Cell. 163:999–1010. 2015. View Article : Google Scholar : PubMed/NCBI

19 

Zhou J, Wan J, Gao X, Zhang X, Jaffrey SR and Qian SB: Dynamic m6A mRNA methylation directs translational control of heat shock response. Nature. 526:591–594. 2015. View Article : Google Scholar : PubMed/NCBI

20 

Xiao W, Adhikari S, Dahal U, Chen YS, Hao YJ, Sun BF, Sun HY, Li A, Ping XL, Lai WY, et al: Nuclear m6A reader YTHDC1 regulates mRNA splicing. Mol Cell. 61:507–519. 2016. View Article : Google Scholar : PubMed/NCBI

21 

Alarcón CR, Lee H, Goodarzi H, Halberg N and Tavazoie SF: N6-methyladenosine marks primary microRNAs for processing. Nature. 519:482–485. 2015. View Article : Google Scholar

22 

Alarcón CR, Goodarzi H, Lee H, Liu X, Tavazoie S and Tavazoie SF: HNRNPA2B1 is a mediator of m6A-dependent nuclear RNA processing events. Cell. 162:1299–1308. 2015. View Article : Google Scholar

23 

Chen T, Hao YJ, Zhang Y, Li MM, Wang M, Han W, Wu Y, Lv Y, Hao J, Wang L, et al: m6A RNA methylation is regulated by microRNAs and promotes reprogramming to pluripotency. Cell Stem Cell. 16:289–301. 2015. View Article : Google Scholar : PubMed/NCBI

24 

Geula S, Moshitch-Moshkovitz S, Dominissini D, Mansour AA, Kol N, Salmon-Divon M, Hershkovitz V, Peer E, Mor N, Manor YS, et al: Stem cells m6A mRNA methylation facilitates resolution of naïve pluripotency toward differentiation. Science. 347:1002–1006. 2015. View Article : Google Scholar : PubMed/NCBI

25 

Patil DP, Chen CK, Pickering BF, Chow A, Jackson C, Guttman M and Jaffrey SR: m6A RNA methylation promotes XIST-mediated transcriptional repression. Nature. 537:369–373. 2016. View Article : Google Scholar : PubMed/NCBI

26 

Xiang Y, Laurent B, Hsu CH, Nachtergaele S, Lu Z, Sheng W, Xu C, Chen H, Ouyang J, Wang S, et al: RNA m6A methylation regulates the ultraviolet-induced DNA damage response. Nature. 543:573–576. 2017. View Article : Google Scholar : PubMed/NCBI

27 

Zhang SY, Zhang SW, Liu L, Meng J and Huang Y: m6A-Driver: Identifying context-specific mRNA m6A methylation-driven gene interaction networks. PLOS Comput Biol. 12:e10052872016. View Article : Google Scholar

28 

Franken NA, Rodermond HM, Stap J, Haveman J and van Bree C: Clonogenic assay of cells in vitro. Nat Protoc. 1:2315–2319. 2006. View Article : Google Scholar

29 

Huang W, Sherman BT and Lempicki RA: Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc. 4:44–57. 2009. View Article : Google Scholar

30 

Szklarczyk D, Morris JH, Cook H, Kuhn M, Wyder S, Simonovic M, Santos A, Doncheva NT, Roth A, Bork P, et al: The STRING database in 2017: Quality-controlled protein-protein association networks, made broadly accessible. Nucleic Acids Res. 45(D1): D362–D368. 2017. View Article : Google Scholar

31 

Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, Amin N, Schwikowski B and Ideker T: Cytoscape: A software environment for integrated models of biomolecular interaction networks. Genome Res. 13:2498–2504. 2003. View Article : Google Scholar : PubMed/NCBI

32 

Bader GD and Hogue CW: An automated method for finding molecular complexes in large protein interaction networks. BMC Bioinformatics. 4:22003. View Article : Google Scholar : PubMed/NCBI

33 

Tempero MA, Malafa MP, Al-Hawary M, Asbun H, Bain A, Behrman SW, Benson AB III, Binder E, Cardin DB, Cha C, et al: Pancreatic adenocarcinoma, version 2.2017, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Cancer Netw. 15:1028–1061. 2017. View Article : Google Scholar

34 

Dhillon AS, Hagan S, Rath O and Kolch W: MAP kinase signalling pathways in cancer. Oncogene. 26:3279–3290. 2007. View Article : Google Scholar : PubMed/NCBI

35 

Corre I, Paris F and Huot J: The p38 pathway, a major pleiotropic cascade that transduces stress and metastatic signals in endothelial cells. Oncotarget. 8:55684–55714. 2017.PubMed/NCBI

36 

Gong F and Miller KM: Mammalian DNA repair: HATs and HDACs make their mark through histone acetylation. Mutat Res. 750:23–30. 2013. View Article : Google Scholar : PubMed/NCBI

37 

Shan W, Jiang Y, Yu H, Huang Q, Liu L, Guo X, Li L, Mi Q, Zhang K and Yang Z: HDAC2 overexpression correlates with aggressive clinicopathological features and DNA-damage response pathway of breast cancer. Am J Cancer Res. 7:1213–1226. 2017.PubMed/NCBI

38 

Livingstone C: IGF2 and cancer. Endocr Relat Cancer. 20:R321–R339. 2013. View Article : Google Scholar : PubMed/NCBI

39 

Cargnello M and Roux PP: Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev. 75:50–83. 2011. View Article : Google Scholar : PubMed/NCBI

40 

Moens U, Kostenko S and Sveinbjørnsson B: The role of mitogen-activated protein kinase-activated protein kinases (MAPKAPKs) in inflammation. Genes (Basel). 4:101–133. 2013. View Article : Google Scholar

41 

Koltai T: Clusterin: A key player in cancer chemoresistance and its inhibition. Onco Targets Ther. 7:447–456. 2014. View Article : Google Scholar : PubMed/NCBI

42 

Albany C and Hahn NM: Heat shock and other apoptosis-related proteins as therapeutic targets in prostate cancer. Asian J Androl. 16:359–363. 2014. View Article : Google Scholar : PubMed/NCBI

43 

Somasagara RR, Spencer SM, Tripathi K, Clark DW, Mani C, Madeira da Silva L, Scalici J, Kothayer H, Westwell AD, Rocconi RP, et al: RAD6 promotes DNA repair and stem cell signaling in ovarian cancer and is a promising therapeutic target to prevent and treat acquired chemoresistance. Oncogene. Aug 14–2017.Epub ahead of print. View Article : Google Scholar : PubMed/NCBI

44 

Rui L, Yuan M, Frantz D, Shoelson S and White MF: SOCS-1 and SOCS-3 block insulin signaling by ubiquitin-mediated degradation of IRS1 and IRS2. J Biol Chem. 277:42394–42398. 2002. View Article : Google Scholar : PubMed/NCBI

45 

Beaurivage C, Champagne A, Tobelaim WS, Pomerleau V, Menendez A and Saucier C: SOCS1 in cancer: An oncogene and a tumor suppressor. Cytokine. 82:87–94. 2016. View Article : Google Scholar : PubMed/NCBI

46 

Cano F, Rapiteanu R, Sebastiaan Winkler G and Lehner PJ: A non-proteolytic role for ubiquitin in deadenylation of MHC-I mRNA by the RNA-binding E3-ligase MEX-3C. Nat Commun. 6:86702015. View Article : Google Scholar : PubMed/NCBI

47 

Burrell RA, McClelland SE, Endesfelder D, Groth P, Weller MC, Shaikh N, Domingo E, Kanu N, Dewhurst SM, Gronroos E, et al: Replication stress links structural and numerical cancer chromosomal instability. Nature. 494:492–496. 2013. View Article : Google Scholar : PubMed/NCBI

48 

Eblen ST: Regulation of chemoresistance via alternative messenger RNA splicing. Biochem Pharmacol. 83:1063–1072. 2012. View Article : Google Scholar : PubMed/NCBI

49 

Zhang J and Manley JL: Misregulation of pre-mRNA alternative splicing in cancer. Cancer Discov. 3:1228–1237. 2013. View Article : Google Scholar : PubMed/NCBI

50 

Savage KI, Gorski JJ, Barros EM, Irwin GW, Manti L, Powell AJ, Pellagatti A, Lukashchuk N, McCance DJ, McCluggage WG, et al: Identification of a BRCA1-mRNA splicing complex required for efficient DNA repair and maintenance of genomic stability. Mol Cell. 54:445–459. 2014. View Article : Google Scholar : PubMed/NCBI

51 

Perry WL III, Shepard RL, Sampath J, Yaden B, Chin WW, Iversen PW, Jin S, Lesoon A, O'Brien KA, Peek VL, et al: Human splicing factor SPF45 (RBM17) confers broad multidrug resistance to anticancer drugs when overexpressed - a phenotype partially reversed by selective estrogen receptor modulators. Cancer Res. 65:6593–6600. 2005. View Article : Google Scholar : PubMed/NCBI

52 

Correa BR, de Araujo PR, Qiao M, Burns SC, Chen C, Schlegel R, Agarwal S, Galante PA and Penalva LO: Functional genomics analyses of RNA-binding proteins reveal the splicing regulator SNRPB as an oncogenic candidate in glioblastoma. Genome Biol. 17:1252016. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

February 2018
Volume 52 Issue 2

Print ISSN: 1019-6439
Online ISSN:1791-2423

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Taketo, K., Konno, M., Asai, A., Koseki, J., Toratani, M., Satoh, T. ... Ogawa, K. (2018). The epitranscriptome m6A writer METTL3 promotes chemo- and radioresistance in pancreatic cancer cells. International Journal of Oncology, 52, 621-629. https://doi.org/10.3892/ijo.2017.4219
MLA
Taketo, K., Konno, M., Asai, A., Koseki, J., Toratani, M., Satoh, T., Doki, Y., Mori, M., Ishii, H., Ogawa, K."The epitranscriptome m6A writer METTL3 promotes chemo- and radioresistance in pancreatic cancer cells". International Journal of Oncology 52.2 (2018): 621-629.
Chicago
Taketo, K., Konno, M., Asai, A., Koseki, J., Toratani, M., Satoh, T., Doki, Y., Mori, M., Ishii, H., Ogawa, K."The epitranscriptome m6A writer METTL3 promotes chemo- and radioresistance in pancreatic cancer cells". International Journal of Oncology 52, no. 2 (2018): 621-629. https://doi.org/10.3892/ijo.2017.4219