Regulatory effects of the long non‑coding RNA RP11‑543N12.1 and microRNA‑324‑3p axis on the neuronal apoptosis induced by the inflammatory reactions of microglia

Cai,Miao; Wang,Yan‑Wen; Xu,Shan‑Hu; Qiao,Song; Shu,Qin‑Fen; Du,Jian‑Zong; Li,Ya‑Guo; Liu,Xiao‑Li

doi:10.3892/ijmm.2018.3736

Regulatory effects of the long non‑coding RNA RP11‑543N12.1 and microRNA‑324‑3p axis on the neuronal apoptosis induced by the inflammatory reactions of microglia

Authors:
- Miao Cai
- Yan‑Wen Wang
- Shan‑Hu Xu
- Song Qiao
- Qin‑Fen Shu
- Jian‑Zong Du
- Ya‑Guo Li
- Xiao‑Li Liu
View Affiliations

Affiliations: Department of Neurology, Zhejiang Hospital, Hangzhou, Zhejiang 310013, P.R. China
Published online on: June 20, 2018 https://doi.org/10.3892/ijmm.2018.3736
Pages: 1741-1755

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

The present study aimed to examine how the long non‑coding RNA (lncRNA) RP11‑543N12.1 interacted with microRNA (miR)‑324‑3p to modify microglials (MIs)‑induced neuroblastoma cell apoptosis, which may pose benefits to the treatment of Alzhemier's disease (AD). The cell model of AD was established by treating SH‑SY5Y cells with amyloid β (Aβ)25‑35, and MI were acquired using primary cell culture technology. The lncRNAs that were differentially expressed between SH‑SY5Y and control cells were screened through a microarray assay and confirmed via polymerase chain reaction. In addition, overexpression of RP11‑543N12.1 and miR‑324‑3p was established by transfection of SH‑SY5Y cells with pcDNA3.1(+)‑RP11‑543N12.1 and miR‑324‑3p mimics, respectively, while downregulation of RP11‑543N12.1 and miR‑324‑3p was achieved by transfection with RP11‑543N12.1‑small interfering RNA (siRNA) and miR‑324‑3p inhibitor, respectively. The interaction between RP11‑543N12.1 and miR‑324‑3p was confirmed with a dual‑luciferase reporter gene assay. The results revealed that the expression levels of total and phosphorylated tau in SH‑SY5Y cells were significantly elevated following Aβ25‑35 treatment (P<0.05), and RP11‑543N12.1 was found to be differentially expressed between the control and Aβ25‑35‑treated cells (P<0.05). Furthermore, the targeted association of RP11‑543N12.1 and miR‑324‑3p was predicted based on miRDB4.0 and PITA databases, and then validated via the dual‑luciferase reporter gene assay. SH‑SY5Y cells transfected with siRNA or inhibitor, and treated with Aβ25‑35 displayed cellular survival and apoptosis that were similar to the normal levels (P<0.05). Finally, co‑culture of MI and SH‑SY5Y cells transfected with RP11‑543N12.1‑siRNA/miR‑324‑3p inhibitor significantly enhanced cell apoptosis (P<0.05). In conclusion, RP11‑543N12.1 targeted miR‑324‑3p to suppress proliferation and promote apoptosis in the AD cell model, suggesting that RP11‑543N12.1 and miR‑324‑3p may be potential biomarkers and therapeutic targets for AD.

View Figures

View References

1	D'Amore JD, Kajdasz ST, McLellan ME, Bacskai BJ, Stern EA and Hyman BT: In vivo multiphoton imaging of a transgenic mouse model of Alzheimer disease reveals marked thioflavine-S-associated alterations in neurite trajectories. J Neuropathol Exp Neurol. 62:137–145. 2003. View Article : Google Scholar : PubMed/NCBI
2	Yoshiyama Y, Higuchi M, Zhang B, Huang SM, Iwata N, Saido TC, Maeda J, Suhara T, Trojanowski JQ and Lee VM: Synapse loss and microglial activation precede tangles in a P301S tauopathy mouse model. Neuron. 53:337–351. 2007. View Article : Google Scholar : PubMed/NCBI
3	Lemere CA, Maier M, Jiang L, Peng Y and Seabrook TJ: Amyloid-beta immunotherapy for the prevention and treatment of Alzheimer disease: Lessons from mice, monkeys, and humans. Rejuvenation Res. 9:77–84. 2006. View Article : Google Scholar : PubMed/NCBI
4	Galimberti D and Scarpini E: Progress in Alzheimer's disease. J Neurol. 259:201–211. 2012. View Article : Google Scholar
5	Alzheimer's Association: 2010 Alzheimer's disease facts and figures. Alzheimers Dement. 6:158–194. 2010. View Article : Google Scholar : PubMed/NCBI
6	Millan MJ: Linking deregulation of non-coding RNA to the core pathophysiology of Alzheimer's disease: An integrative review. Prog Neurobiol. 156:1–68. 2017. View Article : Google Scholar : PubMed/NCBI
7	Qi X, Shao M, Sun H, Shen Y, Meng D and Huo W: Long non-coding RNA SNHG14 promotes microglia activation by regulating miR-145-5p/PLA2G4A in cerebral infarction. Neuroscience. 348:98–106. 2017. View Article : Google Scholar : PubMed/NCBI
8	Lin ST, Heng MY, Ptáček LJ and Fu YH: Regulation of myelination in the central nervous system by nuclear lamin B1 and non-coding RNAs. Transl Neurodegener. 3:42014. View Article : Google Scholar : PubMed/NCBI
9	Liu X, Hou L, Huang W, Gao Y, Lv X and Tang J: The mechanism of long non-coding RNA MEG3 for neurons apoptosis caused by hypoxia: Mediated by miR-181b-12/15-LOX signaling pathway. Front Cell Neuroscie. 10:2012016.
10	Ko CY, Chu YY, Narumiya S, Chi JY, Furuyashiki T, Aoki T, Wang SM, Chang WC and Wang JM: CCAAT/enhancer-binding protein delta/miR135a/thrombospondin 1 axis mediates PGE2-induced angiogenesis in Alzheimer's disease. Neurobiol Aging. 36:1356–1368. 2015. View Article : Google Scholar : PubMed/NCBI
11	Zhao Y, Bhattacharjee S, Jones BM, Dua P, Alexandrov PN, Hill JM and Lukiw WJ: Regulation of TREM2 expression by an NF-κB-sensitive miRNA-34a. Neuroreport. 24:318–323. 2013. View Article : Google Scholar : PubMed/NCBI
12	Jayadev S, Case A, Alajajian B, Eastman AJ, Möller T and Garden GA: Presenilin 2 influences miR146 level and activity in microglia. J Neurochem. 127:592–599. 2013. View Article : Google Scholar : PubMed/NCBI
13	Dharap A, Pokrzywa C, Murali S, Pandi G and Vemuganti R: MicroRNA miR-324-3p induces promoter-mediated expression of RelA gene. PLoS One. 8:e794672013. View Article : Google Scholar : PubMed/NCBI
14	Gutierrez H, O'Keeffe GW, Gavaldà N, Gallagher D and Davies AM: Nuclear factor kappa B signaling either stimulates or inhibits neurite growth depending on the phosphorylation status of p65/RelA. J Neurosci. 28:8246–8256. 2008. View Article : Google Scholar : PubMed/NCBI
15	Peng Y, Gallagher SF, Landmann R, Haines K and Murr MM: The role of p65 NF-kappaB/RelA in pancreatitis-induced Kupffer cell apoptosis. J Gastrointest Surg. 10:837–847. 2006. View Article : Google Scholar : PubMed/NCBI
16	Sheehy AM and Schlissel MS: Overexpression of RelA causes G1 arrest and apoptosis in a pro-B cell line. J Biol Chem. 274:8708–8716. 1999. View Article : Google Scholar : PubMed/NCBI
17	Shankar GM, Li S, Mehta TH, Garcia-Munoz A, Shepardson NE, Smith I, Brett FM, Farrell MA, Rowan MJ, Lemere CA, et al: Amyloid-beta protein dimers isolated directly from Alzheimer's brains impair synaptic plasticity and memory. Nat Med. 14:837–842. 2008. View Article : Google Scholar : PubMed/NCBI
18	McGeer EG and McGeer PL: Inflammatory processes in Alzheimer's disease. Prog Neuropsychopharmacol Biol Psychiatry. 27:741–749. 2003. View Article : Google Scholar : PubMed/NCBI
19	Dobrenis K: Microglia in cell culture and in transplantation therapy for central nervous system disease. Methods. 16:320–344. 1998. View Article : Google Scholar
20	Tan AM, Zhao P, Waxman SG and Hains BC: Early microglial inhibition preemptively mitigates chronic pain development after experimental spinal cord injury. J Rehabil Res Dev. 46:123–133. 2009. View Article : Google Scholar : PubMed/NCBI
21	Roth R, Madhani HD and Garcia JF: Total RNA isolation and quantification of specific RNAs in fission yeast. Methods Mol Biol. 1721:63–72. 2018. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar
23	Mercer TR, Dinger ME and Mattick JS: Long non-coding RNAs: Insights into functions. Nat Rev Genet. 10:155–159. 2009. View Article : Google Scholar : PubMed/NCBI
24	Wu P, Zuo X, Deng H, Liu X, Liu L and Ji A: Roles of long noncoding RNAs in brain development, functional diversification and neurodegenerative diseases. Brain Res Bull. 97:69–80. 2013. View Article : Google Scholar : PubMed/NCBI
25	Batista PJ and Chang HY: Long noncoding RNAs: Cellular address codes in development and disease. Cell. 152:1298–1307. 2013. View Article : Google Scholar : PubMed/NCBI
26	Wang LK, Chen XF, He DD, Li Y and Fu J: Dissection of functional lncRNAs in Alzheimer's disease by construction and analysis of lncRNA-mRNA networks based on competitive endogenous RNAs. Biochem Biophys Res Commun. 485:569–576. 2017. View Article : Google Scholar
27	Ciarlo E, Massone S, Penna I, Nizzari M, Gigoni A, Dieci G, Russo C, Florio T, Cancedda R and Pagano A: An intronic ncRNA-dependent regulation of SORL1 expression affecting Aβ formation is upregulated in post-mortem Alzheimer's disease brain samples. Dis Model Mech. 6:424–433. 2013. View Article : Google Scholar
28	Mus E, Hof PR and Tiedge H: Dendritic BC200 RNA in aging and in Alzheimer's disease. Proc Natl Acad Sci USA. 104:10679–10684. 2007. View Article : Google Scholar : PubMed/NCBI
29	Zhou X and Xu J: Identification of Alzheimer's disease-associated long noncoding RNAs. Neurobiol Aging. 36:2925–2931. 2015. View Article : Google Scholar : PubMed/NCBI
30	Yan B and Wang Z: Long noncoding RNA: Its physiological and pathological roles. DNA Cell Biol. 31(Suppl 1): S34–S41. 2012. View Article : Google Scholar : PubMed/NCBI
31	Xu J, Ai Q, Cao H and Liu Q: MiR-185-3p and miR-324-3p predict radiosensitivity of nasopharyngeal carcinoma and modulate cancer cell growth and apoptosis by targeting SMAD7. Med Sci Monit. 21:2828–2836. 2015. View Article : Google Scholar : PubMed/NCBI
32	Kuo WT, Yu SY, Li SC, Lam HC, Chang HT, Chen WS, Yeh CY, Hung SF, Liu TC, Wu T, et al: MicroRNA-324 in human cancer: miR-324-5p and miR-324-3p have distinct biological functions in human cancer. Anticancer Res. 36:5189–5196. 2016. View Article : Google Scholar : PubMed/NCBI
33	Liu N, Zheng Y, Zhu Y, Xiong S and Chu Y: Selective impairment of CD4+CD25+Foxp3+ regulatory T cells by paclitaxel is explained by Bcl-2/Bax mediated apoptosis. Int Immunopharmacol. 11:212–219. 2011. View Article : Google Scholar
34	Yang Q, Yang K and Li A: microRNA-21 protects against ischemia-reperfusion and hypoxia-reperfusion-induced cardiocyte apoptosis via the phosphatase and tensin homolog/Akt-dependent mechanism. Mol Med Rep. 9:2213–2220. 2014. View Article : Google Scholar : PubMed/NCBI
35	Miyashita T, Krajewski S, Krajewska M, Wang HG, Lin HK, Liebermann DA, Hoffman B and Reed JC: Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene. 9:1799–1805. 1994.PubMed/NCBI
36	Moshrefi M, Spotin A, Kafil HS, Mahami-Oskouei M, Baradaran B, Ahmadpour E and Mansoori B: Tumor suppressor p53 induces apoptosis of host lymphocytes experimentally infected by Leishmania major, by activation of Bax and caspase-3: A possible survival mechanism for the parasite. Parasitol Res. 116:2159–2166. 2017. View Article : Google Scholar : PubMed/NCBI
37	Lukiw WJ: NF-κB-regulated, proinflammatory miRNAs in Alzheimer's disease. Alzheimers Res Ther. 4:472012. View Article : Google Scholar
38	Lee M: Neurotransmitters and microglial-mediated neuroinflammation. Curr Protein Pept Sci. 14:21–32. 2013. View Article : Google Scholar : PubMed/NCBI