METTL14 promotes the migration and invasion of breast cancer cells by modulating N6‑methyladenosine and hsa‑miR‑146a‑5p expression

Yi,Dandan; Wang,Ru; Shi,Xianbiao; Xu,Lei; Yilihamu,Yiminu'er; Sang,Jianfeng

doi:10.3892/or.2020.7515

METTL14 promotes the migration and invasion of breast cancer cells by modulating N6‑methyladenosine and hsa‑miR‑146a‑5p expression

Authors:
- Dandan Yi
- Ru Wang
- Xianbiao Shi
- Lei Xu
- Yiminu'er Yilihamu
- Jianfeng Sang
View Affiliations

Affiliations: Department of General Surgery, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, Jiangsu 210008, P.R. China, Department of General Surgery, Drum Tower Clinical Medical College of Nanjing Medical University, Nanjing, Jiangsu 211166, P.R. China, Department of General Surgery, Drum Tower Clinical Medical College of Nanjing University, Nanjing, Jiangsu 210093, P.R. China
Published online on: February 24, 2020 https://doi.org/10.3892/or.2020.7515
Pages: 1375-1386
Copyright: © Yi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Breast cancer (BC) is the most frequently diagnosed cancer and the leading cause of cancer‑related death among women worldwide. Evidence indicates that posttranscriptional N6‑methyladenosine (m6A) modification modulates BC development. In the present study, we assessed BC and normal tissues to investigate this connection. RNA m6A levels were determined by methylation quantification assay. The effects of methyltransferase‑like 14 (METTL14) gain‑of‑expression or co‑transfection with an m6A inhibitor on cell migration and invasion abilities were determined by Transwell assays. The levels of differentially expressed (DE) miRNAs were verified by real‑time quantitative PCR (RT‑qPCR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes analyses (KEGG) were performed to analyze potential function of target genes of the DE miRNAs. The effects of candidate miRNAs modulated by METTL14 on cell migration and invasion abilities were confirmed by Transwell assays. We demonstrated that m6A methyltransferase METTL14 was significantly upregulated in BC tissues compared with normal tissues. METTL14 gain‑ and loss‑of‑expression regulated m6A levels in MCF‑7 and MDA‑MB‑231 cells. The cell function assays revealed that METTL14 overexpression enhanced the migration and invasion capacities of BC cells. Moreover, treatment with the m6A inhibitor suppressed this enhanced cell migration and invasion. Additionally, aberrant expression of METTL14 reshaped the miRNA profile in BC cell lines. The remodeled DE miRNA/mRNA network was found to be most enriched in cancer pathways, and DE miRNAs were enriched in cell adhesion terms. hsa‑miR‑146a‑5p modulated by METTL14 promoted cell migration and invasion. METTL14 modulates m6A modification and hsa‑miR‑146a‑5p expression, thereby affecting the migration and invasion of breast cancer cells.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar : PubMed/NCBI
2	Nechuta SJ, Caan BJ, Chen WY, Lu W, Chen Z, Kwan ML, Flatt SW, Zheng Y, Zheng W, Pierce JP and Shu XO: Soy food intake after diagnosis of breast cancer and survival: An in-depth analysis of combined evidence from cohort studies of US and Chinese women. Am J Clin Nutr. 23:123–132. 2012. View Article : Google Scholar
3	Zhang T, Li J, He Y, Yang F, Hao Y, Jin W, Wu J, Sun Z, Li Y, Chen Y, et al: A small molecule targeting myoferlin exerts promising anti-tumor effects on breast cancer. Nat Commun. 9:37262018. View Article : Google Scholar : PubMed/NCBI
4	Turner NC, Finn RS, Martin M, Im SA, DeMichele A, Ettl J, Diéras V, Moulder S, Lipatov O, Colleoni M, et al: Clinical considerations of the role of palbociclib in the management of advanced breast cancer patients with and without visceral metastases. Ann Oncol. 29:669–680. 2018. View Article : Google Scholar : PubMed/NCBI
5	Echeverria GV, Powell E, Seth S, Ge Z, Carugo A, Bristow C, Peoples M, Robinson F, Qiu H, Shao J, et al: High-resolution clonal mapping of multi-organ metastasis in triple negative breast cancer. Nat Commun. 9:50792018. View Article : Google Scholar : PubMed/NCBI
6	Machnicka MA, Milanowska K, Osman Oglou O, Purta E, Kurkowska M, Olchowik A, Januszewski W, Kalinowski S, Dunin-Horkawicz S, Rother KM, et al: MODOMICS: A database of RNA modification pathways-2013 update. Nucleic Acids Res. 41 (Database issue):D262–D267. 2013. View Article : Google Scholar : PubMed/NCBI
7	Frye M, Harada BT, Behm M and He C: RNA modifications modulate gene expression during development. Science. 361:1346–1349. 2018. View Article : Google Scholar : PubMed/NCBI
8	Lee M, Kim B and Kim VN: Emerging roles of RNA modification: m(6)A and U-tail. Cell. 158:980–987. 2014. View Article : Google Scholar : PubMed/NCBI
9	Zhao X, Chen Y, Mao Q, Jiang X, Jiang W, Chen J, Xu W, Zhong L and Sun X: Overexpression of YTHDF1 is associated with poor prognosis in patients with hepatocellular carcinoma. Cancer Biomark. 21:859–868. 2018. View Article : Google Scholar : PubMed/NCBI
10	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 methyltransferase. Cell Res. 24:177–189. 2014. View Article : Google Scholar : PubMed/NCBI
11	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
12	Fu Y, Dominissini D, Rechavi G and He C: Gene expression regulation mediated through reversible m⁶A RNA methylation. Nat Rev Genet. 15:293–306. 2014. View Article : Google Scholar : PubMed/NCBI
13	Mathiyalagan P, Adamiak M, Mayourian J, Sassi Y, Liang Y, Agarwal N, Jha D, Zhang S, Kohlbrenner E, Chepurko E, et al: FTO-Dependent m6A Regulates Cardiac Function During Remodeling and Repair. Circulation. 139:518–532. 2018. View Article : Google Scholar
14	Shen DD, Suo FZ, Song QM, Chang J, Zhang T, Hong JJ, Zheng YC and Liu HM: Development of formaldehyde dehydrogenase-coupled assay and antibody-based assays for ALKBH5 activity evaluation. J Pharm Biomed Anal. 162:9–15. 2019. View Article : Google Scholar : PubMed/NCBI
15	Panneerdoss S, Eedunuri VK, Yadav P, Timilsina S, Rajamanickam S, Viswanadhapalli S, Abdelfattah N, Onyeagucha BC, Cui X, Lai Z, et al: Cross-talk among writers, readers, and erasers of m6A regulates cancer growth and progression. Sci Adv. 4:eaar82632018. View Article : Google Scholar : PubMed/NCBI
16	Liu J, Eckert MA, Harada BT, Liu SM, Lu Z, Yu K, Tienda SM, Chryplewicz A, Zhu AC, Yang Y, et al: m⁶A mRNA methylation regulates AKT activity to promote the proliferation and tumorigenicity of endometrial cancer. Nat Cell Biol. 20:1074–1083. 2018. View Article : Google Scholar : PubMed/NCBI
17	Lin S, Choe J, Du P, Triboulet R and Gregory R: The m(6)A methyltransferase METTL3 promotes translation in human cancer cells. Mol Cell. 62:335–345. 2016. View Article : Google Scholar : PubMed/NCBI
18	Zhou S, Bai ZL, Xia D, Zhao ZJ, Zhao R, Wang YY and Zhe H: FTO regulates the chemo-radiotherapy resistance of cervical squamous cell carcinoma (CSCC) by targeting beta-catenin through mRNA demethylation. Mol Carcinog. 57:590–597. 2018. View Article : Google Scholar : PubMed/NCBI
19	Zhang S, Zhao BS, Zhou A, Lin K, Zheng S, Lu Z, Chen Y, Sulman EP, Xie K, Bögler O, et al: m⁶A Demethylase ALKBH5 maintains tumorigenicity of glioblastoma stem-like cells by sustaining FOXM1 expression and cell proliferation program. Cancer Cell. 31:591–606.e596. 2017. View Article : Google Scholar : PubMed/NCBI
20	Cui Q, Shi H, Ye P, Li L, Qu Q, Sun G, Sun G, Lu Z, Huang Y, Yang CG, et al: m⁶A RNA methylation regulates the self-renewal and tumorigenesis of glioblastoma stem cells. Cell Rep. 18:2622–2634. 2017. View Article : Google Scholar : PubMed/NCBI
21	Hiro-Oki I and Yukihide T: The functions of MicroRNAs: mRNA decay and translational repression. Trends Cell Biol. 25:651–665. 2015. View Article : Google Scholar : PubMed/NCBI
22	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 : PubMed/NCBI
23	Cai X, Wang X, Cao C, Gao Y, Zhang S, Yang Z, Liu Y, Zhang X, Zhang W and Ye L: HBXIP-elevated methyltransferase METTL3 promotes the progression of breast cancer via inhibiting tumor suppressor let-7g. Cancer Lett. 28:15652017.
24	Ramayo-Caldas Y, Mach N, Esteve-Codina A, Corominas J, Castelló A, Ballester M, Estellé J, Ibáñez-Escriche N, Fernández AI, Pérez-Enciso M and Folch JM: Liver transcriptome profile in pigs with extreme phenotypes of intramuscular fatty acid composition. BMC Genomics. 13:5472012. View Article : Google Scholar : PubMed/NCBI
25	Kozomara A and Griffiths-Jones S: miRBase: Annotating high confidence microRNAs using deep sequencing data. Nucleic Acids Res. 42((Database issue)): D68–D73. 2014. View Article : Google Scholar : PubMed/NCBI
26	Wright GW and Simon RM: A random variance model for detection of differential gene expression in small microarray experiments. Bioinformatics. 19:2448–2455. 2003. View Article : Google Scholar : PubMed/NCBI
27	Agarwal V, Bell GW, Nam JW and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. Elife. 4:e050052015. View Article : Google Scholar :
28	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
29	Huang da 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 : PubMed/NCBI
30	Hulsegge I, Kommadath A and Smits MA: Globaltest and GOEAST: Two different approaches for Gene Ontology analysis. BMC Proc. 3 (Suppl 4):S102009. View Article : Google Scholar : PubMed/NCBI
31	Kanehisa M and Goto S: KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 28:27–30. 2000. View Article : Google Scholar : PubMed/NCBI
32	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. View Article : Google Scholar : PubMed/NCBI
33	Cheng M, Sheng L, Gao Q, Xiong Q, Zhang H, Wu M, Liang Y, Zhu F, Zhang Y, Zhang X, et al: The mA methyltransferase METTL3 promotes bladder cancer progression via AFF4/NF-κB/MYC signaling network. Oncogene. 38:3667–3680. 2019. View Article : Google Scholar : PubMed/NCBI
34	Sökeland G and Schumacher U: The functional role of integrins during intra- and extravasation within the metastatic cascade. Mol Cancer. 18:122019. View Article : Google Scholar : PubMed/NCBI
35	Massalha S and Weihs D: Metastatic breast cancer cells adhere strongly on varying stiffness substrates, initially without adjusting their morphology. Biomech Model Mechanobiol. 16:961–970. 2017. View Article : Google Scholar : PubMed/NCBI
36	Erson-Bensan AE and Begik O: m6A Modification and Implications for microRNAs. Microrna. 6:97–101. 2017. View Article : Google Scholar : PubMed/NCBI
37	Ma JZ, Yang F, Zhou CC, Liu F, Yuan JH, Wang F, Wang TT, Xu QG, Zhou WP and Sun SH: METTL14 suppresses the metastatic potential of hepatocellular carcinoma by modulating N -methyladenosine-dependent primary MicroRNA processing. Hepatology. 65:529–543. 2017. View Article : Google Scholar : PubMed/NCBI
38	Wang C, Zhang W, Zhang L, Chen X, Liu F, Zhang J, Guan S, Sun Y, Chen P, Wang D, et al: miR-146a-5p mediates epithelial-mesenchymal transition of oesophageal squamous cell carcinoma via targeting Notch2. Br J Cancer. 118:e122018. View Article : Google Scholar : PubMed/NCBI
39	Hsieh JY, Huang TS, Cheng SM, Lin WS, Tsai TN, Lee OK and Wang HW: miR-146a-5p circuitry uncouples cell proliferation and migration, but not differentiation, in human mesenchymal stem cells. Nucleic Acids Res. 41:9753–9763. 2013. View Article : Google Scholar : PubMed/NCBI
40	Gao W, Hua J, Jia Z, Ding J, Han Z, Dong Y, Lin Q and Yao Y: Expression of miR-146a-5p in breast cancer and its role in proliferation of breast cancer cells. Oncol Lett. 15:9884–9888. 2018.PubMed/NCBI