LncRNA ZNRD1-AS1 Promotes Malignant Lung Cell Proliferation, Migration, and Angiogenesis Via the MiR-942/TNS1 Axis and is Positively Regulated by the MA Reader YTHDC2

Overview

Journal Mol Cancer

Publisher Biomed Central

Specialties Molecular Biology
Oncology

Date 2022 Dec 29

PMID 36581942

Authors

Jin Wang

Lirong Tan

Xueting Yu

Xiyuan Cao

Beibei Jia

Rui Chen

Jianxiang Li

Affiliations

Soon will be listed here.

Abstract

Rationale: Lung cancer is the most prevalent form of cancer and has a high mortality rate, making it a global public health concern. The N-methyladenosine (mA) modification is a highly dynamic and reversible process that is involved in a variety of essential biological processes. Using in vitro, in vivo, and multi-omics bioinformatics, the present study aims to determine the function and regulatory mechanisms of the long non-coding (lnc)RNA zinc ribbon domain-containing 1-antisense 1 (ZNRD1-AS1).

Methods: The RNAs that were bound to the mA 'reader' were identified using YTH domain-containing 2 (YTHDC2) RNA immunoprecipitation (RIP)-sequencing. Utilizing methylated RIP PCR/quantitative PCR, pull-down, and RNA stability assays, mA modification and ZNRD1-AS1 regulation were analyzed. Using bioinformatics, the expression levels and clinical significance of ZNRD1-AS1 in lung cancer were evaluated. Using fluorescent in situ hybridization and quantitative PCR assays, the subcellular location of ZNRD1-AS1 was determined. Using cell migration, proliferation, and angiogenesis assays, the biological function of ZNRD1-AS1 in lung cancer was determined. In addition, the tumor suppressor effect of ZNRD1-AS1 in vivo was validated using a xenograft animal model. Through bioinformatics analysis and in vitro assays, the downstream microRNAs (miRs) and competing endogenous RNAs were also predicted and validated.

Results: This study provided evidence that mA modification mediates YTHDC2-mediated downregulation of ZNRD1-AS1 in lung cancer and cigarette smoke-exposed cells. Low levels of ZNRD1-AS1 expression were linked to adverse clinicopathological characteristics, immune infiltration, and prognosis. ZNRD1-AS1 overexpression was shown to suppress lung cancer cell proliferation, migration, and angiogenesis in vitro and in vivo, and to reduce tumor growth in nude mice. ZNRD1-AS1 expression was shown to be controlled by treatment of cells with either the methylation inhibitor 3-Deazaadenosine or the demethylation inhibitor Meclofenamic. Furthermore, the miR-942/tensin 1 (TNS1) axis was demonstrated to be the downstream regulatory signaling pathway of ZNRD1-AS1.

Conclusions: ZNRD1-AS1 serves an important function and has clinical relevance in lung cancer. In addition, the findings suggested that mA modification could mediate the regulation of the ZNRD1-AS1/miR-942/TNS1 axis via the mA reader YTHDC2.

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References

Chen H, Duncan I, Bozorgchami H, Lo S . Tensin1 and a previously undocumented family member, tensin2, positively regulate cell migration. Proc Natl Acad Sci U S A. 2002; 99(2):733-8. PMC: 117374. DOI: 10.1073/pnas.022518699. View

Flanders W, Lally C, Zhu B, Henley S, Thun M . Lung cancer mortality in relation to age, duration of smoking, and daily cigarette consumption: results from Cancer Prevention Study II. Cancer Res. 2003; 63(19):6556-62. View

Dunn G, Old L, Schreiber R . The immunobiology of cancer immunosurveillance and immunoediting. Immunity. 2004; 21(2):137-48. DOI: 10.1016/j.immuni.2004.07.017. View

Wang M, Yang C, Liu X, Zheng J, Xue Y, Ruan X . An upstream open reading frame regulates vasculogenic mimicry of glioma via ZNRD1-AS1/miR-499a-5p/ELF1/EMI1 pathway. J Cell Mol Med. 2020; 24(11):6120-6136. PMC: 7294115. DOI: 10.1111/jcmm.15217. View

Dominissini D, Moshitch-Moshkovitz S, Schwartz S, Salmon-Divon M, Ungar L, Osenberg S . Topology of the human and mouse m6A RNA methylomes revealed by m6A-seq. Nature. 2012; 485(7397):201-6. DOI: 10.1038/nature11112. View

Guo L, Wen J, Han J, Jiang J, Xie S, Feng X . Expression quantitative trait loci in long non-coding RNA ZNRD1-AS1 influence cervical cancer development. Am J Cancer Res. 2015; 5(7):2301-7. PMC: 4548342. View

Du H, Sun J, Chen Z, Nie J, Tong J, Li J . Cigarette smoke-induced failure of apoptosis resulting in enhanced neoplastic transformation in human bronchial epithelial cells. J Toxicol Environ Health A. 2012; 75(12):707-20. DOI: 10.1080/15287394.2012.690088. View

Statello L, Guo C, Chen L, Huarte M . Gene regulation by long non-coding RNAs and its biological functions. Nat Rev Mol Cell Biol. 2020; 22(2):96-118. PMC: 7754182. DOI: 10.1038/s41580-020-00315-9. View

Fatica A, Bozzoni I . Long non-coding RNAs: new players in cell differentiation and development. Nat Rev Genet. 2013; 15(1):7-21. DOI: 10.1038/nrg3606. View

10.

Sato M, Larsen J, Lee W, Sun H, Shames D, Dalvi M . Human lung epithelial cells progressed to malignancy through specific oncogenic manipulations. Mol Cancer Res. 2013; 11(6):638-50. PMC: 3687022. DOI: 10.1158/1541-7786.MCR-12-0634-T. View

11.

Ratnadiwakara M, Anko M . mRNA Stability Assay Using transcription inhibition by Actinomycin D in Mouse Pluripotent Stem Cells. Bio Protoc. 2021; 8(21):e3072. PMC: 8342049. DOI: 10.21769/BioProtoc.3072. View

12.

Wang J, Jiang C, Li N, Wang F, Xu Y, Shen Z . The circEPSTI1/mir-942-5p/LTBP2 axis regulates the progression of OSCC in the background of OSF via EMT and the PI3K/Akt/mTOR pathway. Cell Death Dis. 2020; 11(8):682. PMC: 7443145. DOI: 10.1038/s41419-020-02851-w. View

13.

Li D, Song L, Wen Z, Li X, Jie J, Wang Y . Strong evidence for LncRNA ZNRD1-AS1, and its functional Cis- eQTL locus contributing more to the susceptibility of lung cancer. Oncotarget. 2016; 7(24):35813-35817. PMC: 5094964. DOI: 10.18632/oncotarget.8411. View

14.

Chen Y, Li C, Pan Y, Han S, Feng B, Gao Y . The Emerging Role and Promise of Long Noncoding RNAs in Lung Cancer Treatment. Cell Physiol Biochem. 2016; 38(6):2194-206. DOI: 10.1159/000445575. View

15.

Heguy A, Harvey B, Leopold P, Dolgalev I, Raman T, Crystal R . Responses of the human airway epithelium transcriptome to in vivo injury. Physiol Genomics. 2006; 29(2):139-48. DOI: 10.1152/physiolgenomics.00167.2006. View

16.

An P, Goedert J, Donfield S, Buchbinder S, Kirk G, Detels R . Regulatory variation in HIV-1 dependency factor ZNRD1 associates with host resistance to HIV-1 acquisition. J Infect Dis. 2014; 210(10):1539-48. PMC: 4215072. DOI: 10.1093/infdis/jiu291. View

17.

Yin X, Zhang K, Wang J, Zhou X, Zhang C, Song X . RNA polymerase I subunit 12 plays opposite roles in cell proliferation and migration. Biochem Biophys Res Commun. 2021; 560:112-118. DOI: 10.1016/j.bbrc.2021.04.091. View

18.

Rousseaux S, Debernardi A, Jacquiau B, Vitte A, Vesin A, Nagy-Mignotte H . Ectopic activation of germline and placental genes identifies aggressive metastasis-prone lung cancers. Sci Transl Med. 2013; 5(186):186ra66. PMC: 4818008. DOI: 10.1126/scitranslmed.3005723. View

19.

Cheng C, Wu Y, Xiao T, Xue J, Sun J, Xia H . METTL3-mediated mA modification of ZBTB4 mRNA is involved in the smoking-induced EMT in cancer of the lung. Mol Ther Nucleic Acids. 2021; 23:487-500. PMC: 7806951. DOI: 10.1016/j.omtn.2020.12.001. View

20.

Okayama A, Miyagi Y, Oshita F, Ito H, Nakayama H, Nishi M . Identification of Tyrosine-Phosphorylated Proteins Upregulated during Epithelial-Mesenchymal Transition Induced with TGF-β. J Proteome Res. 2015; 14(10):4127-36. DOI: 10.1021/acs.jproteome.5b00082. View