Long noncoding RNA HCG18 inhibits the differentiation of human bone marrow-derived mesenchymal stem cells in osteoporosis by targeting miR-30a-5p/NOTCH1 axis.
HCG18
Human BMSCs
NOTCH1
Osteoporosis
miR-30a-5p
Journal
Molecular medicine (Cambridge, Mass.)
ISSN: 1528-3658
Titre abrégé: Mol Med
Pays: England
ID NLM: 9501023
Informations de publication
Date de publication:
11 11 2020
11 11 2020
Historique:
received:
11
02
2020
accepted:
14
09
2020
entrez:
12
11
2020
pubmed:
13
11
2020
medline:
11
8
2021
Statut:
epublish
Résumé
Recent studies have demonstrated that long non-coding RNAs (LncRNAs) can influence bone cell differentiation and formation. However, it is unclear whether lncRNA HCG18 is involved in osteoporosis (OP). This study was conducted to investigate the regulation of HCG18 in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). BMSCs were isolated and cultured from mouse pathological models and osteoporosis patients. RT-qPCR was performed to detect the expression of HCG18 and miR-30a-5p in BMSCs. The interaction between HCG18 and miR-30a-5p was analyzed by dual luciferase assay and RNA pulldown assay. The interaction between miR-30a-5p and NOTCH1 3'-UTR was analyzed by dual luciferase assay. RT-qPCR and Western blotting were used to detect the expression of osteogenic genes Runx2, OCN and OPN. Hindlimb-unloaded (HU) mice model was established, and HCG18 was knocked down on bone-formation surfaces by using lentivirus mediated shRNA transfection. The expression of HCG18 was increased in BMSCs of OP patients, while the expression of miR-30a-5p was decreased. The expression of HCG18 and miR-30a-5p was negatively correlated in BMSCs. During the differentiation from BMSCs to osteoblasts, the expression of HCG18 was significantly downregulated, and the expression of miR-30a-5p was significantly upregulated. Overexpression of HCG18 was able to reverse the osteogenic-induced upregulation of miR-30a-5p expression, and knockdown of HCG18 further promoted the expression of miR-30a-5p. In addition, miR-30a-5p partially abolished the effect of HCG18 on osteogenic differentiation of BMSCs. NOTCH1 was a target protein of miR-30a-5p, and upregulation of NOTCH1 reversed the effect of miR-30a-5p on osteogenic differentiation of BMSCs. Furthermore, this study found that lentivirus mediated HCG18 knockdown on the bone-formation surfaces of hindlimb-unloaded (HU) mice partially alleviated unloading-induced bone loss CONCLUSIONS: HCG18 inhibited osteogenic differentiation of BMSCs induced by OP via the miR-30a-5p/NOTCH1 axis. HCG18 can be identified as a regulator of osteogenic differentiation of BMSCs.
Sections du résumé
BACKGROUND
Recent studies have demonstrated that long non-coding RNAs (LncRNAs) can influence bone cell differentiation and formation. However, it is unclear whether lncRNA HCG18 is involved in osteoporosis (OP). This study was conducted to investigate the regulation of HCG18 in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).
METHODS
BMSCs were isolated and cultured from mouse pathological models and osteoporosis patients. RT-qPCR was performed to detect the expression of HCG18 and miR-30a-5p in BMSCs. The interaction between HCG18 and miR-30a-5p was analyzed by dual luciferase assay and RNA pulldown assay. The interaction between miR-30a-5p and NOTCH1 3'-UTR was analyzed by dual luciferase assay. RT-qPCR and Western blotting were used to detect the expression of osteogenic genes Runx2, OCN and OPN. Hindlimb-unloaded (HU) mice model was established, and HCG18 was knocked down on bone-formation surfaces by using lentivirus mediated shRNA transfection.
RESULTS
The expression of HCG18 was increased in BMSCs of OP patients, while the expression of miR-30a-5p was decreased. The expression of HCG18 and miR-30a-5p was negatively correlated in BMSCs. During the differentiation from BMSCs to osteoblasts, the expression of HCG18 was significantly downregulated, and the expression of miR-30a-5p was significantly upregulated. Overexpression of HCG18 was able to reverse the osteogenic-induced upregulation of miR-30a-5p expression, and knockdown of HCG18 further promoted the expression of miR-30a-5p. In addition, miR-30a-5p partially abolished the effect of HCG18 on osteogenic differentiation of BMSCs. NOTCH1 was a target protein of miR-30a-5p, and upregulation of NOTCH1 reversed the effect of miR-30a-5p on osteogenic differentiation of BMSCs. Furthermore, this study found that lentivirus mediated HCG18 knockdown on the bone-formation surfaces of hindlimb-unloaded (HU) mice partially alleviated unloading-induced bone loss CONCLUSIONS: HCG18 inhibited osteogenic differentiation of BMSCs induced by OP via the miR-30a-5p/NOTCH1 axis. HCG18 can be identified as a regulator of osteogenic differentiation of BMSCs.
Identifiants
pubmed: 33176682
doi: 10.1186/s10020-020-00219-6
pii: 10.1186/s10020-020-00219-6
pmc: PMC7656763
doi:
Substances chimiques
Biomarkers
0
MIRN301 microRNA, mouse
0
MicroRNAs
0
Notch1 protein, mouse
0
RNA, Long Noncoding
0
Receptor, Notch1
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
106Références
Calcif Tissue Int. 2017 Aug;101(2):111-131
pubmed: 28324124
Sci Rep. 2017 Oct 16;7(1):13234
pubmed: 29038477
Mol Med Rep. 2018 Mar;17(3):4554-4560
pubmed: 29344643
J Proteome Res. 2011 Mar 4;10(3):1416-9
pubmed: 21186791
J Cell Biochem. 2018 Mar;119(3):2843-2850
pubmed: 29068476
J Cell Biochem. 2018 Nov;119(10):7971-7981
pubmed: 29236315
BMJ Open. 2018 Mar 1;8(3):e015187
pubmed: 29500198
Int J Obes (Lond). 2018 Jun;42(6):1140-1150
pubmed: 29899524
Oncogene. 2013 Aug 15;32(33):3915-21
pubmed: 22986530
Biomed Pharmacother. 2017 May;89:1178-1186
pubmed: 28320084
DNA Cell Biol. 2019 Apr;38(4):341-351
pubmed: 30839226
Hum Cell. 2014 Oct;27(4):151-61
pubmed: 24573839
J Bone Miner Res. 2011 Feb;26(2):317-30
pubmed: 20740684
Mod Pathol. 2010 Jun;23(6):814-23
pubmed: 20348879
J Bone Miner Res. 2004 Feb;19(2):235-44
pubmed: 14969393
Curr Osteoporos Rep. 2017 Apr;15(2):110-119
pubmed: 28303448
J Biomed Inform. 2011 Oct;44(5):839-47
pubmed: 21605702
Sci Rep. 2015 Dec 21;5:18655
pubmed: 26686902
Nat Rev Cancer. 2017 Mar;17(3):145-159
pubmed: 28154375
Bosn J Basic Med Sci. 2017 Nov 20;17(4):295-301
pubmed: 29055350
Mol Cell Biochem. 2013 Apr;376(1-2):189-95
pubmed: 23358924
Arch Osteoporos. 2017 Dec;12(1):43
pubmed: 28425085
Mol Cell Biochem. 2014 Jul;392(1-2):85-93
pubmed: 24752351
PLoS One. 2014 May 13;9(5):e97123
pubmed: 24824427
Cell Biol Int. 2006 Sep;30(9):681-7
pubmed: 16870478
Nucleic Acids Res. 2015 Apr 20;43(7):3712-25
pubmed: 25779046
Cancer Lett. 2012 Feb 1;315(1):28-37
pubmed: 22055459
Ann Intern Med. 2017 Jun 6;166(11):818-839
pubmed: 28492856
RNA. 2015 Aug;21(8):1433-43
pubmed: 26078267
Methods Mol Biol. 2016;1402:271-286
pubmed: 26721498
Front Immunol. 2017 Oct 16;8:1312
pubmed: 29085370
Sci Rep. 2018 Jun 14;8(1):9127
pubmed: 29904151
Bone. 2014 May;62:22-8
pubmed: 24508387
Hum Genet. 2019 Feb;138(2):151-166
pubmed: 30661131
Blood. 2017 Mar 2;129(9):1124-1133
pubmed: 28115368
Retrovirology. 2014 Dec 13;11:118
pubmed: 25496667
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
World J Gastroenterol. 2014 Jan 7;20(1):126-32
pubmed: 24415865
Gene. 2018 Jun 5;658:28-35
pubmed: 29518546
Neurol Res. 2010 Mar;32(2):148-56
pubmed: 19473555
Stem Cells. 2008 Aug;26(8):2075-82
pubmed: 18499895
Eur J Neurosci. 2005 Mar;21(6):1469-77
pubmed: 15845075
J Bone Miner Res. 1997 Jan;12(1):16-23
pubmed: 9240721
J Cell Mol Med. 2011 Nov;15(11):2297-306
pubmed: 21143388
Biosci Trends. 2018 Jul 17;12(3):275-281
pubmed: 29794404
Cell Death Dis. 2018 Feb 7;9(2):176
pubmed: 29416009
RNA. 2010 Feb;16(2):324-37
pubmed: 20026622