https://doi.org/10.4081/ejh.2022.3438

Long non-coding RNA WAC antisense RNA 1 mediates hepatitis B virus replication in vitro by reinforcing miR-192-5p/ATG7-induced autophagy

Vol. 66 No. 3 (2022)
Submitted: 22 May 2022
Accepted: 21 July 2022
Published: 2 September 2022
hepatitis B virus, lncRNA WAC-AS1, miR-192-5p/AGT7 axis, autophagy
Abstract Views: 904
PDF: 540
HTML: 16
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Authors

Minkai Cao
Department of Obstetrics and Gynecology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China.
Deping Yuan
Department of Obstetrics and Gynecology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China.
Hongxiu Jiang
Department of Obstetrics and Gynecology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China.
Guanlun Zhou
Department of Obstetrics and Gynecology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China.
Chao Chen
Department of Obstetrics and Gynecology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China.
Guorong Han
njyy033@njucm.edu.cn
https://orcid.org/0000-0002-1397-8185
Department of Obstetrics and Gynecology, the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China.

Long non-coding RNA WAC antisense RNA 1 (lncRNA WAC-AS1) is involved in the replication of the hepatitis B virus (HBV). The purpose of this study was to determine its functions and specific mechanism. The levels of lncRNA WAC-AS1, RNA (miR)-192-5p and were examined in serum of HBV-infected patients and in HepG2.2.15 cells using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. Using the database starBase, the target binding sites of lncRNA WAC-AS1 and miR-192-5p were predicted and confirmed by dual-luciferase reporter assay and RNA pull-down assay. The expression of pgRNA and HBV DNA was determined by qRT-PCR, while the levels of HBeAg and HBsAg were measured by enzyme-linked immunosorbent assay (ELISA). Using laser scanning confocal microscopy, the light chain 3 (LC3) expression was analyzed. qRT-PCR and Western blotting were used to assess the expression of beclin-1, p62, and LC3I/II. Overexpression of lncRNA WAC-AS1, upregulation of ATG7. and downregulation of miR-192-5p were observed in the serum of HBV-infected patients and the in vitro model. miR-192-5p directly targets lncRNA WAC-AS1. LncRNA WAC-AS1 was downregulated in lncRNA WAC-AS1-shRNA‒transfected cells. miR-192-5p was upregulated in lncRNA WAC-AS1-shRNA-transfected cells and downregulated in cells transfected with a miR-192-5p inhibitor. In HepG2 2.15 cells, the downregulation of lncRNA WAC-AS1 inhibited HBV replication and autophagy. In contrast, the miR-192-5p inhibitor-transfected group exhibited the opposite results, and ATG7 overexpression reversed the effects of miR-192-5p mimic or lncRNA WAC-AS1-shRNA on HBV replication and cell autophagy. Our findings indicate that lncRNA WAC-AS1 regulates HBV replication by reinforcing the autophagy induced by miR-192-5p/ATG7. Consequently, lncRNA WAC-AS1 may serve as a therapeutically-promising target in HBV patients.

Dimensions

Altmetric

PlumX Metrics

Downloads

Download data is not yet available.

Citations

Crossref
Scopus
Google Scholar
Europe PMC
Li Q, Lomonosova E, Donlin MJ, Cao F, O'Dea A, Milleson B, et al. Amide-containing alpha-hydroxytropolones as inhibitors of hepatitis B virus replication. Antiviral Res 2020;177:104777. DOI: https://doi.org/10.1016/j.antiviral.2020.104777
Liu Y, Veeraraghavan V, Pinkerton M, Fu J, Douglas MW, George J, et al. Viral biomarkers for hepatitis B virus-related hepatocellular carcinoma occurrence and recurrence. Front Microbiol 2021;12:665201. DOI: https://doi.org/10.3389/fmicb.2021.665201
Cao LH, Zhao PL, Liu ZM, Sun SC, Xu DB, Zhang JD, et al. Efficacy and safety of nucleoside analogues in preventing vertical transmission of the hepatitis B virus from father to infant. Genet Mol Res 2015;14:15539-46. DOI: https://doi.org/10.4238/2015.December.1.4
Feng J, Yang G, Liu Y, Gao Y, Zhao M, Bu Y, et al. LncRNA PCNAP1 modulates hepatitis B virus replication and enhances tumor growth of liver cancer. Theranostics 2019;9:5227-45. DOI: https://doi.org/10.7150/thno.34273
Zheng J, Zhou Z, Qiu Y, Wang M, Yu H, Wu Z, et al. A prognostic ferroptosis-related lncRNAs signature associated with immune landscape and radiotherapy response in glioma. Front Cell Dev Biol 2021;9:675555. DOI: https://doi.org/10.3389/fcell.2021.675555
Xia X, Zhang H, Xia P, Zhu Y, Liu J, Xu K, et al. Identification of Glycolysis-related lncRNAs and the novel lncRNA WAC-AS1 promotes glycolysis and tumor progression in hepatocellular carcinoma. Front Oncol 2021;11:733595. DOI: https://doi.org/10.3389/fonc.2021.733595
Li X, Guo Y, Wang X, Ge A, Wang H, Fan K, et al. Clinical significance of serum miR-487b in HBV-related hepatocellular carcinoma and its potential mechanism. Infect Dis (Lond) 2021;53:546-54. DOI: https://doi.org/10.1080/23744235.2021.1901981
Dong RF, Zhuang YJ, Wang Y, Zhang ZY, Xu XZ, Mao YR, et al. Tumor suppressor miR-192-5p targets TRPM7 and inhibits proliferation and invasion in cervical cancer. Kaohsiung J Med Sci 2021;37:699-708. DOI: https://doi.org/10.1002/kjm2.12398
Chen X, Su X, Lin M, Fu B, Zhou C, Ling C, et al. Expression of miR-192-5p in colon cancer serum and its relationship with clinicopathologic features. Am J Transl Res 2021;13:9371-6.
Liu XL, Cao HX, Wang BC, Xin FZ, Zhang RN, Zhou D, et al. miR-192-5p regulates lipid synthesis in non-alcoholic fatty liver disease through SCD-1. World J Gastroenterol 2017;23:8140-51. DOI: https://doi.org/10.3748/wjg.v23.i46.8140
Livak and Schmittgen. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods 2001;25:402-8. DOI: https://doi.org/10.1006/meth.2001.1262
Xie M, Yang Z, Liu Y, Zheng M. The role of HBV-induced autophagy in HBV replication and HBV related-HCC. Life Sci 2018;205:107-12. DOI: https://doi.org/10.1016/j.lfs.2018.04.051
Wang D, Yan X, Zhang M, Ren C, Wang L, Ma J, et al. Association between liver cirrhosis and estimated glomerular filtration rates in patients with chronic HBV infection. Medicine (Baltimore) 2020;99:e21387. DOI: https://doi.org/10.1097/MD.0000000000021387
Li H, Yan L, Shi Y, Lv D, Shang J, Bai L, et al. Hepatitis B virus infection: Overview. Adv Exp Med Biol 2020;1179:1-16. DOI: https://doi.org/10.1007/978-981-13-9151-4_1
Han GR, Jiang HX, Wang CM, Ding Y, Wang GJ, Yue X, et al. Long-term safety and efficacy of telbivudine in infants born to mothers treated during the second or third trimesters of pregnancy. J Viral Hepat 2017;24:514-21. DOI: https://doi.org/10.1111/jvh.12670
Pan CQ, Duan Z, Dai E, Zhang S, Han G, Wang Y, et al. Tenofovir to prevent hepatitis B transmission in mothers with high viral load. N Engl J Med 2016;374:2324-34. DOI: https://doi.org/10.1056/NEJMoa1508660
Shahen SM, Elshenawy SZ, Mohamed SE, Talaat RM. Genetic polymorphisms in the miR-372 (rs12983273) and LncRNA HULC (rs7763881) genes and susceptibility to hepatitis B virus (HBV) infection. Mol Biol Rep 2021;48:7901-6. DOI: https://doi.org/10.1007/s11033-021-06818-8
Ren F, Ren JH, Song CL, Tan M, Yu HB, Zhou YJ, et al. LncRNA HOTAIR modulates hepatitis B virus transcription and replication by enhancing SP1 transcription factor. Clin Sci (Lond. 2020;134:3007-22. DOI: https://doi.org/10.1042/CS20200970
Li F, Deng Y, Zhang S, Zhu B, Wang J, Wang J, et al. Human hepatocyte-enriched miRNA-192-3p promotes HBV replication through inhibiting Akt/mTOR signalling by targeting ZNF143 in hepatic cell lines. Emerg Microbes Infect 2022;11:616-28. DOI: https://doi.org/10.1080/22221751.2022.2037393
Wang J, Chen J, Liu Y, Zeng X, Wei M, Wu S, et al. Hepatitis B virus induces autophagy to promote its replication by the axis of miR-192-3p-XIAP through NF kappa B signaling. Hepatology 2019;69:974-92. DOI: https://doi.org/10.1002/hep.30248
Tang C, Yuan P, Wang J, Zhang Y, Chang X, Jin D, et al. MiR-192-5p regulates the proliferation and apoptosis of cholangiocarcinoma cells by activating MEK/ERK pathway. 3 Biotech 2021;11:99. DOI: https://doi.org/10.1007/s13205-021-02650-w
Fu S, Ma C, Tang X, Ma X, Jing G, Zhao N, et al. MiR-192-5p inhibits proliferation, migration, and invasion in papillary thyroid carcinoma cells by regulation of SH3RF3. Biosci Rep 2021;41:BSR20210342. DOI: https://doi.org/10.1042/BSR20210342
Chen L, Ming X, Li W, Bi M, Yan B, Wang X, et al. The microRNA-155 mediates hepatitis B virus replication by reinforcing SOCS1 signalling-induced autophagy. Cell Biochem Funct 2020;38:436-42. DOI: https://doi.org/10.1002/cbf.3488
Wang X, Lin Y, Liu S, Zhu Y, Lu K, Broering R, Lu M. O-GlcNAcylation modulates HBV replication through regulating cellular autophagy at multiple levels. FASEB J 2020;34:14473-89. DOI: https://doi.org/10.1096/fj.202001168RR
Lou L, Tian M, Chang J, Li F, Zhang G. MiRNA-192-5p attenuates airway remodeling and autophagy in asthma by targeting MMP-16 and ATG7. Biomed Pharmacother 2020;122:109692. DOI: https://doi.org/10.1016/j.biopha.2019.109692
Zhou W, Yu Q, Ma J, Xu C, Wu D, Li C. Triamcinolone acetonide combined with 5-fluorouracil suppresses urethral scar fibroblasts autophagy and fibrosis by increasing miR-192-5p expression. Am J Transl Res 2021;13:5956-68.
Zhou P, Dong M, Wang J, Li F, Zhang J, Gu J. Baseline serum miR-125b levels predict virologic response to nucleos(t)ide analogue treatment in patients with HBeAg-positive chronic hepatitis B. Exp Ther Med 2018;16:3805-12. DOI: https://doi.org/10.3892/etm.2018.6685
Zhao L, Wang B, Sun L, Sun B, Li Y. Association of miR-192-5p with Atherosclerosis and its Effect on Proliferation and Migration of Vascular Smooth Muscle Cells. Mol Biotechnol 2021;63:1244-51. DOI: https://doi.org/10.1007/s12033-021-00376-x
Wang S, Sun Y, Wang Y, Wang A, Kou B, Che Y, et al. ASPP2 inhibits hepatitis B virus replication by preventing nucleus translocation of HSF1 and attenuating the transactivation of ATG7. J Cell Mol Med 2021;25:6899-908. DOI: https://doi.org/10.1111/jcmm.16699
Collier JJ, Suomi F, Oláhová M, McWilliams TG, Taylor RW. Emerging roles of ATG7 in human health and disease. EMBO Mol Med 2021;13:e14824. DOI: https://doi.org/10.15252/emmm.202114824

Ethics Approval

The experimental protocols were approved by the Ethics Committee of the Second Hospital of Nanjing, Nanjing University of Chinese Medicine, Nanjing, China

Rights

Chinese National Research Grant of the Thirteenth Five-Year Plan for the Key Projects in Infectious Diseases; Development Project of The Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University

How to Cite

Cao, M., Yuan, D., Jiang, H., Zhou, G., Chen, C., & Han, G. (2022). Long non-coding RNA WAC antisense RNA 1 mediates hepatitis B virus replication <em>in vitro</em> by reinforcing miR-192-5p/ATG7-induced autophagy. European Journal of Histochemistry, 66(3). https://doi.org/10.4081/ejh.2022.3438
Copyright (c) 2022 The Author(s) Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

PAGEPress has chosen to apply the Creative Commons Attribution NonCommercial 4.0 International License (CC BY-NC 4.0) to all manuscripts to be published.