open access

Vol 26, No 5 (2019)
Original articles — Basic science and experimental cardiology
Published online: 2018-05-08
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The long noncoding RNA THRIL knockdown protects hypoxia-induced injuries of H9C2 cells through regulating miR-99a

Jingwen Xia, Nianxin Jiang, Yansong Li, Yong Wei, Xuan Zhang
DOI: 10.5603/CJ.a2018.0054
·
Pubmed: 29745968
·
Cardiol J 2019;26(5):564-574.

open access

Vol 26, No 5 (2019)
Original articles — Basic science and experimental cardiology
Published online: 2018-05-08

Abstract

Background: Myocardial infarction (MI) is a leading cause of disease with high morbidity and mortality worldwide. Recent studies have revealed that long non-coding RNAs (lncRNAs) are involved in
heart disease pathogenesis. This study aimed to investigate the effect and the molecular basis of THRIL on hypoxia-injured H9C2 cells.


Methods: THRIL, miR-99a and Brahma-related gene 1 (Brg1) expressions in H9C2 cells were altered by transient transfections. The cells were subjected to hypoxia for 4 h, and then the levels of THRIL, miR-99a and Brg1 were investigated. Cell viability, migration and invasion, and apoptotic cells were respectively measured by trypan blue exclusion assay, transwell migration assay and flow cytometry assay. Dual luciferase reporter assay was conducted to verify the interaction between miR-99a and THRIL. Furthermore, levels of apoptosis-, PI3K/AKT and mTOR pathways-related factors were measured by western blotting.


Results: Hypoxia induced an increase of THRIL but a reduction of miR-99a and Brg1. THRIL inhibition significantly attenuated hypoxia-induced cell injuries, as increased cell viability, migration and invasion, and decreased cell apoptosis. THRIL negatively regulated miR-99a expression through sponging with miR-99a binding site, and miR-99a inhibition abolished the protective effects of THRIL knockdown against hypoxia-induced injury in H9C2 cells. Furthermore, miR-99a positively regulated the expression of Brg1. Brg1 inhibition promoted hypoxia-induced cell injuries, while Brg1 overexpression alleviated hypoxia-induced cell injuries. Moreover, Brg1 overexpression activated PI3K/AKT and mTOR pathways.


Conclusions: This study demonstrated that THRIL inhibition represented a protective effect against
hypoxia-induced injuries in H9C2 cells by up-regulating miR-99a expression.

Abstract

Background: Myocardial infarction (MI) is a leading cause of disease with high morbidity and mortality worldwide. Recent studies have revealed that long non-coding RNAs (lncRNAs) are involved in
heart disease pathogenesis. This study aimed to investigate the effect and the molecular basis of THRIL on hypoxia-injured H9C2 cells.


Methods: THRIL, miR-99a and Brahma-related gene 1 (Brg1) expressions in H9C2 cells were altered by transient transfections. The cells were subjected to hypoxia for 4 h, and then the levels of THRIL, miR-99a and Brg1 were investigated. Cell viability, migration and invasion, and apoptotic cells were respectively measured by trypan blue exclusion assay, transwell migration assay and flow cytometry assay. Dual luciferase reporter assay was conducted to verify the interaction between miR-99a and THRIL. Furthermore, levels of apoptosis-, PI3K/AKT and mTOR pathways-related factors were measured by western blotting.


Results: Hypoxia induced an increase of THRIL but a reduction of miR-99a and Brg1. THRIL inhibition significantly attenuated hypoxia-induced cell injuries, as increased cell viability, migration and invasion, and decreased cell apoptosis. THRIL negatively regulated miR-99a expression through sponging with miR-99a binding site, and miR-99a inhibition abolished the protective effects of THRIL knockdown against hypoxia-induced injury in H9C2 cells. Furthermore, miR-99a positively regulated the expression of Brg1. Brg1 inhibition promoted hypoxia-induced cell injuries, while Brg1 overexpression alleviated hypoxia-induced cell injuries. Moreover, Brg1 overexpression activated PI3K/AKT and mTOR pathways.


Conclusions: This study demonstrated that THRIL inhibition represented a protective effect against
hypoxia-induced injuries in H9C2 cells by up-regulating miR-99a expression.

Get Citation

Keywords

THRIL; miR-99a; Brg1; myocardial infarction; hypoxia

About this article
Title

The long noncoding RNA THRIL knockdown protects hypoxia-induced injuries of H9C2 cells through regulating miR-99a

Journal

Cardiology Journal

Issue

Vol 26, No 5 (2019)

Pages

564-574

Published online

2018-05-08

DOI

10.5603/CJ.a2018.0054

Pubmed

29745968

Bibliographic record

Cardiol J 2019;26(5):564-574.

Keywords

THRIL
miR-99a
Brg1
myocardial infarction
hypoxia

Authors

Jingwen Xia
Nianxin Jiang
Yansong Li
Yong Wei
Xuan Zhang

References (36)
  1. Hoffmann R. The potential for reduction of educational differences in ischaemic heart disease mortality in Europe: The role of three lifestyle risk factors. Eur J Public Health. 2016: ckw104.
  2. Ren Li, Liu W, Wang Y, et al. Investigation of hypoxia-induced myocardial injury dynamics in a tissue interface mimicking microfluidic device. Anal Chem. 2013; 85(1): 235–244.
  3. Peng W, Si S, Zhang Q, et al. Long non-coding RNA MEG3 functions as a competing endogenous RNA to regulate gastric cancer progression. J Exp Clin Cancer Res. 2015; 34: 79.
  4. Zhou RM, Wang XQ, Yao J, et al. Identification and characterization of proliferative retinopathy-related long noncoding RNAs. Biochem Biophys Res Commun. 2015; 465(3): 324–330.
  5. Vausort M, Wagner DR, Devaux Y. Long noncoding RNAs in patients with acute myocardial infarction. Circ Res. 2014; 115(7): 668–677.
  6. Haemmerle M, Gutschner T. Long non-coding RNAs in cancer and development: where do we go from here? Int J Mol Sci. 2015; 16(1): 1395–1405.
  7. Hung CL, Wang LY, Yu YL, et al. A long noncoding RNA connects c-Myc to tumor metabolism. Proc Natl Acad Sci U S A. 2014; 111(52): 18697–18702.
  8. Uchida S, Dimmeler S. Long noncoding RNAs in cardiovascular diseases. Circ Res. 2015; 116(4): 737–750.
  9. Li Z, Chao TC, Chang KY, et al. The long noncoding RNA THRIL regulates TNFα expression through its interaction with hnRNPL. Proc Natl Acad Sci U S A. 2014; 111(3): 1002–1007.
  10. Hsieh TC, Wijeratne EK, Liang JY, et al. Differential control of growth, cell cycle progression, and expression of NF-kappaB in human breast cancer cells MCF-7, MCF-10A, and MDA-MB-231 by ponicidin and oridonin, diterpenoids from the chinese herb Rabdosia rubescens. Biochem Biophys Res Commun. 2005; 337(1): 224–231.
  11. Itou J, Oishi I, Kawakami H, et al. Migration of cardiomyocytes is essential for heart regeneration in zebrafish. Development. 2012; 139(22): 4133–4142.
  12. Hang CT, Yang J, Han P, et al. Chromatin regulation by Brg1 underlies heart muscle development and disease. Nature. 2010; 466(7302): 62–67.
  13. Chakrabarti M, Banik NL, Ray SK. Photofrin based photodynamic therapy and miR-99a transfection inhibited FGFR3 and PI3K/Akt signaling mechanisms to control growth of human glioblastoma In vitro and in vivo. PLoS One. 2013; 8(2): e55652.
  14. Yang Z, Han Y, Cheng K, et al. miR-99a directly targets the mTOR signalling pathway in breast cancer side population cells. Cell Prolif. 2014; 47(6): 587–595.
  15. Liu B, Che W, Xue J, et al. SIRT4 prevents hypoxia-induced apoptosis in H9c2 cardiomyoblast cells. Cell Physiol Biochem. 2013; 32(3): 655–662.
  16. Gong C, Maquat LE. lncRNAs transactivate STAU1-mediated mRNA decay by duplexing with 3' UTRs via Alu elements. Nature. 2011; 470(7333): 284–288.
  17. Kanduri C. Kcnq1ot1: a chromatin regulatory RNA. Semin Cell Dev Biol. 2011; 22(4): 343–350.
  18. Wang K, Liu CY, Zhou LY, et al. APF lncRNA regulates autophagy and myocardial infarction by targeting miR-188-3p. Nat Commun. 2015; 6: 6779.
  19. Clemson CM, Hutchinson JN, Sara SA, et al. An architectural role for a nuclear noncoding RNA: NEAT1 RNA is essential for the structure of paraspeckles. Mol Cell. 2009; 33(6): 717–726.
  20. Liao J, He Q, Li M, et al. LncRNA MIAT: Myocardial infarction associated and more. Gene. 2016; 578(2): 158–161.
  21. Wang P, Fu H, Cui J, et al. Differential lncRNA‑mRNA co‑expression network analysis revealing the potential regulatory roles of lncRNAs in myocardial infarction. Mol Med Rep. 2016; 13(2): 1195–1203.
  22. Yang KC, Yamada KA, Patel AY, et al. Deep RNA sequencing reveals dynamic regulation of myocardial noncoding RNAs in failing human heart and remodeling with mechanical circulatory support. Circulation. 2014; 129(9): 1009–1021.
  23. Grote P, Wittler L, Hendrix D, et al. The tissue-specific lncRNA Fendrr is an essential regulator of heart and body wall development in the mouse. Dev Cell. 2013; 24(2): 206–214.
  24. Sang KL, Na KL, Yong CL. 150 Long Non-Coding RNA Thril and Hnrnpl Regulates Helicobacter pylori cagA Induced-Inflammation by Inhibition of NF-κB Translocation. Gastroenterology. 2016; 150(4): S37.
  25. Salmena L, Poliseno L, Tay Y, et al. A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language? Cell. 2011; 146(3): 353–358.
  26. Liu XH, Sun M, Nie FQ, et al. Lnc RNA HOTAIR functions as a competing endogenous RNA to regulate HER2 expression by sponging miR-331-3p in gastric cancer. Mol Cancer. 2014; 13: 92.
  27. Li Q, Xie J, Wang B, et al. Overexpression of microRNA-99a Attenuates Cardiac Hypertrophy. PLoS One. 2016; 11(2): e0148480.
  28. Li Q, Xie J, Li R, et al. Overexpression of microRNA-99a attenuates heart remodelling and improves cardiac performance after myocardial infarction. J Cell Mol Med. 2014; 18(5): 919–928.
  29. Yang SY, Wang YQ, Gao HM, et al. The clinical value of circulating miR-99a in plasma of patients with acute myocardial infarction. Eur Rev Med Pharmacol Sci. 2016; 20(24): 5193–5197.
  30. Xiao C, Gao Lu, Hou Yu, et al. Chromatin-remodelling factor Brg1 regulates myocardial proliferation and regeneration in zebrafish. Nat Commun. 2016; 7: 13787.
  31. Hang CT, Yang J, Han P, et al. Chromatin regulation by Brg1 underlies heart muscle development and disease. Nature. 2010; 466(7302): 62–67.
  32. Bi Y, Wang G, Liu X, et al. Low-after-high glucose down-regulated Cx43 in H9c2 cells by autophagy activation via cross-regulation by the PI3K/Akt/mTOR and MEK/ERK signal pathways. Endocrine. 2017; 56(2): 336–345.
  33. Han D, Wan C, Liu F, et al. Jujuboside A Protects H9C2 Cells from Isoproterenol-Induced Injury via Activating PI3K/Akt/mTOR Signaling Pathway. Evid Based Complement Alternat Med. 2016; 2016: 9593716.
  34. Wang M, Sun Gb, Sun X, et al. Cardioprotective effect of salvianolic acid B against arsenic trioxide-induced injury in cardiac H9c2 cells via the PI3K/Akt signal pathway. Toxicol Lett. 2013; 216(2-3): 100–107.
  35. Zhang ZL, Fan Y, Liu ML. Ginsenoside Rg1 inhibits autophagy in H9c2 cardiomyocytes exposed to hypoxia/reoxygenation. Mol Cell Biochem. 2012; 365(1-2): 243–250.
  36. Li J, He Z, Liu R, Ma M, Yang L, Ye C. Brg1 promotes osteogenic differentiation of mesenchymal stem cells by regulating Runx2-mediated Wnt and PI3K/AKT pathways.

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