Overexpression of lncRNA Dancr inhibits apoptosis and enhances autophagy to protect cardiomyocytes from endoplasmic reticulum stress injury via sponging microRNA-6324

Li,Jiong; Xie,Jing; Wang,Yan-Zhen; Gan,Yi-Rong; Wei,Ling; Ding,Guan-Waner; Ding,Yan-Hong; Xie,Ding-Xiong

doi:10.3892/mmr.2020.11755

Overexpression of lncRNA Dancr inhibits apoptosis and enhances autophagy to protect cardiomyocytes from endoplasmic reticulum stress injury via sponging microRNA-6324

Authors:
- Jiong Li
- Jing Xie
- Yan-Zhen Wang
- Yi-Rong Gan
- Ling Wei
- Guan-Waner Ding
- Yan-Hong Ding
- Ding-Xiong Xie
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Affiliations: Key Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou, Gansu 730000, P.R. China, Department of Ultrasonic Diagnosis, The First People's Hospital of Lanzhou, Lanzhou, Gansu 730050, P.R. China, Gansu Cardiovascular Institute, Lanzhou, Gansu 730050, P.R. China, Outpatient Department, The First People's Hospital of Lanzhou, Lanzhou, Gansu 730050, P.R. China, Medical Department, Shijiazhuang People's Medical College, Shijiazhuang, Hebei 050599, P.R. China, Anesthesiology Department, The First People's Hospital of Lanzhou, Lanzhou, Gansu 730050, P.R. China
Published online on: December 3, 2020 https://doi.org/10.3892/mmr.2020.11755
Article Number: 116
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Endoplasmic reticulum stress (ERS) contributes to the pathogenesis of myocardial ischemia/reperfusion injury and myocardial infarction (MI). Long non-coding RNAs (lncRNAs) serve an important role in cardiovascular diseases, and lncRNA discrimination antagonizing non-protein coding RNA (Dancr) alleviates cardiomyocyte damage. microRNA (miR)-6324 was upregulated in MI model rats and was predicted to bind to Dancr. The present study aimed to investigate the role of Dancr in ERS-induced cardiomyocytes and the potential underlying mechanisms. Tunicamycin (Tm) was used to induce ERS. Cell viability, apoptosis and levels of associated proteins, ERS and autophagy in Dancr-overexpression H9C2 cells and miR-6234 mimic-transfected H9C2 cells were assessed using Cell Counting Kit-8, TUNEL staining and western blot assay, respectively. The results suggested that Dancr expression levels and cell viability were downregulated by Tm in a concentration-dependent manner compared with the control group. Tm induced apoptosis, ERS and autophagy, as indicated by an increased ratio of apoptotic cells, increased expression levels of Bax, cleaved (c)-caspase-3/9, glucose-regulated protein 78 kDa (GRP78), phosphorylated (p)-inositol-requiring enzyme-1α (IRE1α), spliced X-box-binding protein 1 (Xbp1s), IRE1α, activating transcription factor (ATF)6, ATF4, Beclin 1 and microtubule associated protein 1 light chain 3α (LC3)II/I, and decreased expression levels of Bcl-2, unspliced Xbp1 (Xbp1u) and p62 in the Tm group compared with the control group. Moreover, the results indicated that compared with the Tm + overexpression (Oe)-negative control (NC) group, the Tm + Oe-Dancr group displayed decreased apoptosis, but enhanced ERS and autophagy to restore cellular homeostasis. Compared with the Tm + Oe-NC group, the Tm + Oe-Dancr group decreased the ratio of apoptotic cells, decreased expression levels of Bax, c-caspase-3/9 and Xbp1u, and increased expression levels of Bcl-2, p-IRE1α, Xbp1s, Beclin 1 and LC3II/I. Dancr overexpression also significantly downregulated miR-6324 expression compared with Oe-NC. The dual-luciferase reporter assay further indicated an interaction between Dancr and miR-6324. In addition, miR-6324 mimic partially reversed the effects of Dancr overexpression on Tm-induced apoptosis, ERS and autophagy. In conclusion, lncRNA Dancr overexpression protected cardiomyocytes against ERS injury via sponging miR-6324, thus inhibiting apoptosis, enhancing autophagy and restoring ER homeostasis.

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1	Moran AE, Roth GA, Narula J and Mensah GA: 1990-2010 global cardiovascular disease atlas. Glob Heart. 9:3–16. 2014. View Article : Google Scholar : PubMed/NCBI
2	Mehta LS, Beckie TM, DeVon HA, Grines CL, Krumholz HM, Johnson MN, Lindley KJ, Vaccarino V, Wang TY, Watson KE, et al American Heart Association Cardiovascular Disease in Women and Special Populations Committee of the Council on Clinical Cardiology, Council on Epidemiology and Preventionm Council on Cardiovascular and Stroke Nursing, Council on Quality of Care and Outcomes Research, : Acute Myocardial Infarction in Women: A Scientific Statement From the American Heart Association. Circulation. 133:916–947. 2016. View Article : Google Scholar : PubMed/NCBI
3	Reed GW, Rossi JE and Cannon CP: Acute myocardial infarction. Lancet. 389:197–210. 2017. View Article : Google Scholar : PubMed/NCBI
4	Tamis-Holland JE, Jneid H, Reynolds HR, Agewall S, Brilakis ES, Brown TM, Lerman A, Cushman M, Kumbhani DJ, Arslanian-Engoren C, et al American Heart Association Interventional Cardiovascular Care Committee of the Council on Clinical Cardiology; Council on Cardiovascular and Stroke Nursing; Council on Epidemiology and Prevention; and Council on Quality of Care and Outcomes Research, : Contemporary Diagnosis and Management of Patients With Myocardial Infarction in the Absence of Obstructive Coronary Artery Disease: A Scientific Statement From the American Heart Association. Circulation. 139:e891–e908. 2019. View Article : Google Scholar : PubMed/NCBI
5	Heusch G and Gersh BJ: The pathophysiology of acute myocardial infarction and strategies of protection beyond reperfusion: A continual challenge. Eur Heart J. 38:774–784. 2017.PubMed/NCBI
6	Asaria P, Elliott P, Douglass M, Obermeyer Z, Soljak M, Majeed A and Ezzati M: Acute myocardial infarction hospital admissions and deaths in England: A national follow-back and follow-forward record-linkage study. Lancet Public Health. 2:e191–e201. 2017. View Article : Google Scholar : PubMed/NCBI
7	Dall C, Khan M, Chen CA and Angelos MG: Oxygen cycling to improve survival of stem cells for myocardial repair: A review. Life Sci. 153:124–131. 2016. View Article : Google Scholar : PubMed/NCBI
8	Jarroux J, Morillon A and Pinskaya M: History, Discovery, and Classification of lncRNAs. Adv Exp Med Biol. 1008:1–46. 2017. View Article : Google Scholar : PubMed/NCBI
9	Deniz E and Erman B: Long noncoding RNA (lincRNA), a new paradigm in gene expression control. Funct Integr Genomics. 17:135–143. 2017. View Article : Google Scholar : PubMed/NCBI
10	Zhao Z, Sun W, Guo Z, Zhang J, Yu H and Liu B: Mechanisms of lncRNA/microRNA interactions in angiogenesis. Life Sci. 254:1169002020. View Article : Google Scholar : PubMed/NCBI
11	Sallam T, Sandhu J and Tontonoz P: Long noncoding RNA discovery in cardiovascular disease: Decoding Form to Function. Circ Res. 122:155–166. 2018. View Article : Google Scholar : PubMed/NCBI
12	Ballantyne MD, McDonald RA and Baker AH: lncRNA/MicroRNA interactions in the vasculature. Clin Pharmacol Ther. 99:494–501. 2016. View Article : Google Scholar : PubMed/NCBI
13	Thin KZ, Liu X, Feng X, Raveendran S and Tu JC: LncRNA-DANCR: A valuable cancer related long non-coding RNA for human cancers. Pathol Res Pract. 214:801–805. 2018. View Article : Google Scholar : PubMed/NCBI
14	Yuan SX, Wang J, Yang F, Tao QF, Zhang J, Wang LL, Yang Y, Liu H, Wang ZG, Xu QG, et al: Long noncoding RNA DANCR increases stemness features of hepatocellular carcinoma by derepression of CTNNB1. Hepatology. 63:499–511. 2016. View Article : Google Scholar : PubMed/NCBI
15	Guo L, Gu J, Hou S, Liu D, Zhou M, Hua T, Zhang J, Ge Z and Xu J: Long non-coding RNA DANCR promotes the progression of non-small-cell lung cancer by inhibiting p21 expression. OncoTargets Ther. 12:135–146. 2018. View Article : Google Scholar
16	Bai Y, Zhang G, Chu H, Li P and Li J: The positive feedback loop of lncRNA DANCR/miR-138/Sox4 facilitates malignancy in non-small cell lung cancer. Am J Cancer Res. 9:270–284. 2019.PubMed/NCBI
17	Wang Y, Zeng X, Wang N, Zhao W, Zhang X, Teng S, Zhang Y and Lu Z: Long noncoding RNA DANCR, working as a competitive endogenous RNA, promotes ROCK1-mediated proliferation and metastasis via decoying of miR-335-5p and miR-1972 in osteosarcoma. Mol Cancer. 17:892018. View Article : Google Scholar : PubMed/NCBI
18	Qiu L, Zhao Q, Dai L, Zhu A, Xu X, Zhao S and Chen J: Long non-coding RNA DANCR alleviates hypoxia-caused H9c2 cells damage through up regulation of HIF-1α. Artif Cells Nanomed Biotechnol. 48:533–541. 2020. View Article : Google Scholar : PubMed/NCBI
19	Zhang M, Tang M, Wu Q, Wang Z, Chen Z, Ding H, Hu X, Lv X, Zhao S, Sun J, et al: LncRNA DANCR attenuates brain microvascular endothelial cell damage induced by oxygen-glucose deprivation through regulating of miR-33a-5p/XBP1s. Aging (Albany NY). 12:1778–1791. 2020. View Article : Google Scholar : PubMed/NCBI
20	Zuo S, Kong D, Wang C, Liu J, Wang Y, Wan Q, Yan S, Zhang J, Tang J, Zhang Q, et al: CRTH2 promotes endoplasmic reticulum stress-induced cardiomyocyte apoptosis through m-calpain. EMBO Mol Med. 10:e82372018.simplehttps://doi.org/10.15252/emmm.201708237 View Article : Google Scholar : PubMed/NCBI
21	Wang X, Xu L, Gillette TG, Jiang X and Wang ZV: The unfolded protein response in ischemic heart disease. J Mol Cell Cardiol. 117:19–25. 2018. View Article : Google Scholar : PubMed/NCBI
22	Yan M, Shu S, Guo C, Tang C and Dong Z: Endoplasmic reticulum stress in ischemic and nephrotoxic acute kidney injury. Ann Med. 50:381–390. 2018. View Article : Google Scholar : PubMed/NCBI
23	Yamamoto K and Ichikawa S: Tunicamycin: Chemical synthesis and biosynthesis. J Antibiot (Tokyo). 72:924–933. 2019. View Article : Google Scholar : PubMed/NCBI
24	Wang S, Wang Z, Fan Q, Guo J, Galli G, Du G, Wang X and Xiao W: Ginkgolide K protects the heart against endoplasmic reticulum stress injury by activating the inositol-requiring enzyme 1α/X box-binding protein-1 pathway. Br J Pharmacol. 173:2402–2418. 2016. View Article : Google Scholar : PubMed/NCBI
25	Fernández A, Ordóñez R, Reiter RJ, González-Gallego J and Mauriz JL: Melatonin and endoplasmic reticulum stress: Relation to autophagy and apoptosis. J Pineal Res. 59:292–307. 2015. View Article : Google Scholar : PubMed/NCBI
26	Hu J, Huang CX, Rao PP, Cao GQ, Zhang Y, Zhou JP, Zhu LY, Liu MX and Zhang GG: MicroRNA-155 inhibition attenuates endoplasmic reticulum stress-induced cardiomyocyte apoptosis following myocardial infarction via reducing macrophage inflammation. Eur J Pharmacol. 857:1724492019. View Article : Google Scholar : PubMed/NCBI
27	Li X, Zhao J, Geng J, Chen F, Wei Z, Liu C, Zhang X, Li Q, Zhang J, Gao L, et al: Long non-coding RNA MEG3 knockdown attenuates endoplasmic reticulum stress-mediated apoptosis by targeting p53 following myocardial infarction. J Cell Mol Med. 23:8369–8380. 2019. View Article : Google Scholar : PubMed/NCBI
28	Zhao N, Mi L, Zhang X, Xu M, Yu H, Liu Z, Liu X, Guan G, Gao W and Wang J: Enhanced MiR-711 transcription by PPARγ induces endoplasmic reticulum stress-mediated apoptosis targeting calnexin in rat cardiomyocytes after myocardial infarction. J Mol Cell Cardiol. 118:36–45. 2018. View Article : Google Scholar : PubMed/NCBI
29	Xu Y, Wu L, Chen A, Xu C and Feng Q: Protective effects of olive leaf extract on acrolein-exacerbated myocardial infarction via an endoplasmic reticulum stress pathway. Int J Mol Sci. 19:4932018.simplehttps://doi.org/10.3390/ijms19020493 View Article : Google Scholar
30	Cătană CS, Atanasov AG and Berindan-Neagoe I: Natural products with anti-aging potential: Affected targets and molecular mechanisms. Biotechnol Adv. 36:1649–1656. 2018. View Article : Google Scholar : PubMed/NCBI
31	Dong Z, Chu G, Sima Y and Chen G: Djhsp90s are crucial regulators during planarian regeneration and tissue homeostasis. Biochem Biophys Res Commun. 498:723–728. 2018. View Article : Google Scholar : PubMed/NCBI
32	Wang L, Li Y, Ning N, Wang J, Yan Z, Zhang S, Jiao X, Wang X and Liu H: Decreased autophagy induced by β1-adrenoceptor autoantibodies contributes to cardiomyocyte apoptosis. Cell Death Dis. 9:4062018. View Article : Google Scholar : PubMed/NCBI
33	Aisa Z, Liao GC, Shen XL, Chen J, Li L and Jiang SB: Effect of autophagy on myocardial infarction and its mechanism. Eur Rev Med Pharmacol Sci. 21:3705–3713. 2017.PubMed/NCBI
34	Kanamori H, Takemura G, Goto K, Maruyama R, Ono K, Nagao K, Tsujimoto A, Ogino A, Takeyama T, Kawaguchi T, et al: Autophagy limits acute myocardial infarction induced by permanent coronary artery occlusion. Am J Physiol Heart Circ Physiol. 300:H2261–H2271. 2011. View Article : Google Scholar : PubMed/NCBI
35	Wu D, Zhang K and Hu P: The role of autophagy in acute myocardial infarction. Front Pharmacol. 10:5512019. View Article : Google Scholar : PubMed/NCBI
36	Yu Y, Sun G, Luo Y, Wang M, Chen R, Zhang J, Ai Q, Xing N and Sun X: Cardioprotective effects of Notoginsenoside R1 against ischemia/reperfusion injuries by regulating oxidative stress- and endoplasmic reticulum stress- related signaling pathways. Sci Rep. 6:217302016. View Article : Google Scholar : PubMed/NCBI
37	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
38	Liao B, Dong S, Xu Z, Gao F, Zhang S and Liang R: LncRNA Kcnq1ot1 renders cardiomyocytes apoptosis in acute myocardial infarction model by up-regulating Tead1. Life Sci. 256:1178112020. View Article : Google Scholar : PubMed/NCBI
39	Lai L, Liu Y, Liu Y, Zhang N, Cao S, Zhang X and Wu D: Role of endoplasmic reticulum oxidase 1α in H9C2 cardiomyocytes following hypoxia/reoxygenation injury. Mol Med Rep. 22:1420–1428. 2020. View Article : Google Scholar : PubMed/NCBI
40	Walter P and Ron D: The unfolded protein response: From stress pathway to homeostatic regulation. Science. 334:1081–1086. 2011. View Article : Google Scholar : PubMed/NCBI
41	Oyadomari S and Mori M: Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ. 11:381–389. 2004. View Article : Google Scholar : PubMed/NCBI
42	Durante W: Targeting endoplasmic reticulum stress in hypoxia-induced cardiac injury. Vascul Pharmacol. 83:1–3. 2016. View Article : Google Scholar : PubMed/NCBI
43	Levy JMM, Towers CG and Thorburn A: Targeting autophagy in cancer. Nat Rev Cancer. 17:528–542. 2017. View Article : Google Scholar : PubMed/NCBI
44	Lin R, Su Z, Tan X, Su Y, Chen Y, Shu X, Liang S, Wang J and Xie S: Effect of endoplasmic reticulum stress and autophagy in the regulation of post-infarct cardiac eepair. Arch Med Res. 49:576–582. 2018. View Article : Google Scholar : PubMed/NCBI
45	Wiersma M, Meijering RAM, Qi XY, Zhang D, Liu T, Hoogstra-Berends F, Sibon OCM, Henning RH, Nattel S and Brundel BJJM: Endoplasmic reticulum stress is associated with autophagy and cardiomyocyte remodeling in experimental and human atrial fibrillation. J Am Heart Assoc. 6:e0064582017.doi: 10.1161/JAHA.117.006458. View Article : Google Scholar : PubMed/NCBI
46	Holdt LM and Teupser D: Long noncoding RNA ANRIL: Lnc-ing genetic variation at the chromosome 9p21 locus to molecular mechanisms of atherosclerosis. Front Cardiovasc Med. 5:1452018. View Article : Google Scholar : PubMed/NCBI
47	Chi JS, Li JZ, Jia JJ, Zhang T, Liu XM and Yi L: Long non-coding RNA ANRIL in gene regulation and its duality in atherosclerosis. J Huazhong Univ Sci Technolog Med Sci. 37:816–822. 2017.doi: 10.1007/s11596-017-1812-y. PubMed/NCBI
48	Chang JC, Hu WF, Lee WS, Lin JH, Ting PC, Chang HR, Shieh KR, Chen TI and Yang KT: Intermittent hypoxia induces autophagy to protect cardiomyocytes from endoplasmic reticulum stress and apoptosis. Front Physiol. 10:9952019. View Article : Google Scholar : PubMed/NCBI