Top

Published in:

01-12-2019 | Original Research

Long Non-coding RNA XIST May Influence Cervical Ossification of the Posterior Longitudinal Ligament Through Regulation of miR-17-5P/AHNAK/BMP2 Signaling Pathway

Authors: Xinyuan Liao, Dezhi Tang, Haisong Yang, Yu Chen, Deyu Chen, Lianshun Jia, Lili Yang, Xiongsheng Chen

Published in: Calcified Tissue International | Issue 6/2019

Abstract

Long non-coding RNAs (lncRNAs) play an important role in the development of bone-related diseases. This study was conducted to investigate the role and mechanism of lncRNA X inactive specific transcript (XIST) in the occurrence of cervical ossification of the posterior longitudinal ligament (OPLL). Here, primary human ligament fibroblasts cells (LFCs) were isolated from 30 cases of OPLL and 30 normal cervical posterior longitudinal ligament (non-OPLL) tissues to perform the qPCR and Western blot assay. We found that the mRNA level of lncRNA XIST was significantly increased in OPLL LFCs compared to non-OPLL LFCs. By bioinformatics analysis, we found that lncRNA XIST has four binding sites for miR-17-5p and found that the mRNA level of miR-17-5p was also significantly decreased in OPLL LFCs compared to non-OPLL LFCs. Since AHNAK is the target gene of miR-17-5p, we further found that the expression of AHNAK was significantly reduced in non-OPLL LFCs after being transfected with miR-17-5p mimic. The qPCR results showed that the mRNA expressions of BMP2 and Runx2 were significantly decreased. After being transfected with lncRNA XIST siRNA in the non-OPLL LFCs, the mRNA levels of lncRNA XIST, AHNAK, BMP2, and Runx2 were significantly decreased and the phosphorylated protein of Smad1/5/8 was reduced. After being cultured by mechanical vibration, the mRNA levels of lncRNA XIST, AHNAK, BMP2, Runx2, COL1, OC, OPN, and Phospho1 were significantly increased, but the mRNA expression of miR-17-5p was significantly decreased. The expression of phosphorylated Smad1/5/8 protein was also significantly increased. Together, this study was the first to determine that XIST gene inhibition plays an important role in the occurrence of cervical OPLL, through the mechanism of regulation of miR-17-5P/AHNAK/BMP2 signaling pathway. Thus, XIST may be a potential target that could be modulated for the treatment of cervical OPLL.

Xu N, Yu M, Liu X, Sun C, Chen Z, Liu Z (2017) A systematic review of complications in thoracic spine surgery for ossification of the posterior longitudinal ligament. Eur Spine J 26(7):1803–1809CrossRef

Fujimori T, Le H, Hu SS, Chin C, Pekmezci M, Schairer W, Tay BK, Hamasaki T, Yoshikawa H, Iwasaki M (2015) Ossification of the posterior longitudinal ligament of the cervical spine in 3161 patients: a CT-based study. Spine 40(7):E394–E403CrossRef

Li ZS, Zhang GB, Sheng HF et al (1999) Incidence of cervical ossification of the posterior longitudinal ligament in neck and shoulder pain patients in northern China. Chin J Spine Spinal Cord. 9(5):285–286 (in Chinese)

Abiola R, Rubery P, Mesfin A (2016) Ossification of the posterior longitudinal ligament: etiology, diagnosis, and outcomes of nonoperative and operative management. Glob Spine J 6(2):195–204CrossRef

Nakajima M, Takahashi A, Tsuji T et al (2014) A genome-wide association study identifies susceptibility loci for ossification of the posterior longitudinal ligament of the spine. Nat Genet 46(9):1012–1016CrossRef

Tanaka H, Nagai E, Murata H et al (2001) Involvement of bone morphogenetic protein-2 (BMP-2) in the pathological ossification process of the spinal ligament. Rheumayology 40(10):1163–1168CrossRef

Kawaguchi Y, Furushima K, Sugimori K et al (2003) Association between polymorphism of the transforming growth factor-beta1 gene with the radiologic characteristic and ossification of the posterior longitudinal ligament. Spine 28(13):1424–1426PubMed

Sawada T, Kishiya M, Kanemaru K et al (2008) Possible role of extracellular nucleotides in ectopic ossification of human spinal ligaments. J Pharmacol Sci 106(1):152–161CrossRef

Sun M, Kraus WL (2015) From discovery to function: the expanding roles of long noncoding RNAs in physiology and disease. Endocr Rev 36(1):25–64CrossRef

10.

Zhang R, Xia LQ, Lu WW, Zhang J, Zhu JS (2016) LncRNAs and cancer. Oncol Lett 12(2):1233–1239CrossRef

11.

Huynh NP, Anderson BA, Guilak F, McAlinden A (2017) Emerging roles for long noncoding RNAs in skeletal biology and disease. Connect Tissue Res 58(1):116–141CrossRef

12.

Simionescu-Bankston A, Kumar A (2016) Noncoding RNAs in the regulation of skeletal muscle biology in health and disease. J Mol Med 94(8):853–866CrossRef

13.

Nie M, Deng ZL, Liu J, Wang DZ (2015) Emerging roles for long noncoding RNAs in skeletal biology and disease. Biomed Res Int 2015:676575PubMedPubMedCentral

14.

Li L, Liu B, Wapinski OL, Tsai MC, Qu K, Zhang J, Carlson JC, Lin M, Fang F, Gupta RA, Helms JA, Chang HY (2013) Targeted disruption of Hotair leads to homeotic transformation and gene derepression. Cell Rep 5(1):3–12CrossRef

15.

Saydam O, Shen Y, Würdinger T, Senol O, Boke E, James MF, Tannous BA, Stemmer-Rachamimov AO, Yi M, Stephens RM, Fraefel C, Gusella JF, Krichevsky AM, Breakefield XO (2009) Downregulated microRNA-200a in meningiomas promotes tumor growth by reducing E-cadherin and activating the Wnt/beta-catenin signaling pathway. Mol Cell Biol 29(21):5923–5940CrossRef

16.

Zhu L, Xu PC (2013) Downregulated LncRNA-ANCR promotes osteoblast differentiation by targeting EZH2 and regulating Runx2 expression. Biochem Biophys Res Commun 432(4):612–617CrossRef

17.

Li L, Lv G, Wang B, Kuang L (2018) The role of lncRNA XIST/miR-211 axis in modulating the proliferation and apoptosis of osteoarthritis chondrocytes through CXCR1 and MAPK signaling. Biochem Biophys Res Commun 503:2555–2562CrossRef

18.

Chen CK, Blanco M, Jackson C, Aznauryan E, Ollikainen N, Surka C, Chow A, Crerase A, McDonel P, Guttman M (2016) Xist recruits the X chromosome to the nuclear lamina to enable chromosome-wide silencing. Science 354:468–472CrossRef

19.

McHugh CA, Chen CK, Chow A et al (2015) The Xist lncRNA interacts directly with SHARP to silence transcription through HDAC3. Nature 521(7551):232–236CrossRef

20.

Yao Y, Ma J, Xue Y et al (2015) Knockdown of long non-coding RNA XIST exerts tumor-suppressive functions in human glioblastoma stem cells by up-regulating miR-152. Cancer Lett 359(1):75–86CrossRef

21.

Fang J, Sun CC, Gong C (2016) Long noncoding RNA XIST acts as an oncogene in non-small cell lung cancer by epigenetically repressing KLF2 expression. Biochem Biophys Res Commun 478:811–817CrossRef

22.

Song P, Ye LF, Zhang C, Peng T, Zhou XH (2016) Long non-coding RNA XIST exerts oncogenic functions in human nasopharyngeal carcinoma by targeting miR-34a-5p. Gene 592:8–14CrossRef

23.

Zhuang LK, Yang YT, Ma X, Han B, Wang ZS, Zhao QY et al (2016) MicroRNA-92b promotes hepatocellular carcinoma progression by targeting smad7 and is mediated by long non-coding RNA XIST. Cell Death Dis 7:e2203CrossRef

24.

Ren C, Li X, Wang T et al (2015) Functions and mechanisms of long noncoding RNAs in ovarian cancer. Int J Gynecol Cancer 25:566–569CrossRef

25.

Wang Z, Liu M, Zhu H et al (2010) Suppression of p21 by c-myc through members of miR-17 family at the post-transcriptional level. Int J Oncol 37(5):1315–1321CrossRef

26.

Wu Q, Luo G, Yang Z et al (2014) miR-17-5p promotes proliferation by targeting SOCS6 in gastric cancer cells. FEBS Lett 588(12):2055–2062CrossRef

27.

Fan M, Sethuraman A, Brown M et al (2014) Systematic analysis of metastasisassociated genes identifies miR-17-5p as a metastatic suppressor of basal-like breast cancer. Breast Cancer Res Treat 146(3):487–502CrossRef

28.

Fang Y, Xu C, Fu Y (2015) MicroRNA-17-5p induces drug resistance and invasion of ovarian carcinoma cells by targeting PTEN signaling. J Biol Res 22(1):1–10

29.

Li H, Li T, Wang S et al (2013) miR-17-5p and miR-106a are involved in the balance between osteogenic and adipogenic differentiation of adipose-derived mesenchymal stem cells. Stem Cell Rev 10:313–324CrossRef

30.

Fang T, Wu Q, Zhou L et al (2016) miR-106b-5p and miR-17-5p suppress osteogenic differentiation by targeting Smad5 and inhibit bone formation. Exp Cell Res 347:74–82CrossRef

31.

Shin S, Seong JK, Bae YS (2016) Ahnak stimulates BMP2-mediated adipocyte differentiation through Smad1 activation. Obesity 24:398–407CrossRef

32.

Kon T, Yamazaki M, Tagawa M et al (1997) Bone morphogenetic protein-2 stimulates differentiation of cultured spinal ligament cells from patients with ossification of the posterior longitudinal ligament. Calcif Tissue Int 60:291–296CrossRef

33.

Tanno M, Furukawa KI, Ueyama K et al (2003) Uniaxial cyclic stretch induces osteogenic differentiation and synthesis of bone morphogenetic proteins of spinal ligament cells derived from patients with ossification of the posterior longitudinal ligaments. Bone 33:475–484CrossRef

34.

Iwasawa T, Iwasaki K, Sawada T et al (2006) Pathophysiological role of endothelin in ectopic ossification of human spinal ligaments induced by mechanical stress. Calcif Tissue Int 79:422–430CrossRef

35.

Iwasaki K, Furukawa KI, Tanno M et al (2004) Uni-axial cyclic stretch induces Cbfa1 expression in spinal ligament cells derived from patients with ossification of the posterior longitudinal ligament. Calcif Tissue Int 74:448–457CrossRef

36.

Ohishi H, Furukawa K, Iwasaki K et al (2003) Role of prostaglandin I2 in the gene expression induced by mechanical stress in spinal ligament cells derived from patients with ossification of the posterior longitudinal ligament. J Pharmacol Exp Ther 305:818–824CrossRef

37.

Tsukamoto N, Maeda T, Miura H et al (2006) Repetitive tensile stress to rat caudal vertebrae inducing cartilage formation in the spinal ligaments: a possible role of mechanical stress in the development of ossification of the spinal ligaments. Spine 5:234–242PubMed

38.

Ikegawa S (2014) Genomic study of ossification of the posterior longitudinal ligament of the spine. Proc Jpn Acad Ser B Phys Biol Sci 90:405–412CrossRef

39.

Yang HS, Lu XH, Chen DY et al (2011) Upregulated expression of connexin43 in spinal ligament fibroblasts derived from patients presenting ossification of the posterior longitudinal ligament. Spine 36:2267–2274CrossRef

40.

Yang HS, Lu XH, Chen DY et al (2011) Mechanical strain induces Cx43 expression in spinal ligament fibroblasts derived from patients presenting ossification of the posterior longitudinal ligament. Eur Spine J 20:1459–1465CrossRef

Title: Long Non-coding RNA XIST May Influence Cervical Ossification of the Posterior Longitudinal Ligament Through Regulation of miR-17-5P/AHNAK/BMP2 Signaling Pathway
Authors: Xinyuan Liao
Dezhi Tang
Haisong Yang
Yu Chen
Deyu Chen
Lianshun Jia
Lili Yang
Xiongsheng Chen
Publication date: 01-12-2019
Publisher: Springer US
Published in: Calcified Tissue International / Issue 6/2019
Print ISSN: 0171-967X
Electronic ISSN: 1432-0827
DOI: https://doi.org/10.1007/s00223-019-00608-y

ACC 2024 Congress Coverage

Heart Failure Video

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Long Non-coding RNA XIST May Influence Cervical Ossification of the Posterior Longitudinal Ligament Through Regulation of miR-17-5P/AHNAK/BMP2 Signaling Pathway

Abstract

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 6/2019

Effects of Elevated Parathyroid Hormone Levels on Muscle Health, Postural Stability and Quality of Life in Vitamin D-Insufficient Healthy Women: A Cross-Sectional Study

Associations of Smoking and Alcohol and Coffee Intake with Fracture and Bone Mineral Density: A Mendelian Randomization Study

Functional Measures of Sarcopenia: Prevalence, and Associations with Functional Disability in 10,892 Adults Aged 65 Years and Over from Six Lower- and Middle-Income Countries

Understanding Patients’ Perspectives and Educational Needs by Type of Osteoporosis in Men and Women and People with Glucocorticosteroid-Induced Osteoporosis: A Qualitative Study to Improve Disease Management

Multiple Fractures and Impaired Bone Fracture Healing in a Patient with Pycnodysostosis and Hypophosphatasia

Substrate Strain Mitigates Effects of β-Aminopropionitrile-Induced Reduction in Enzymatic Crosslinking

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page