Changed cellular functions and aberrantly expressed miRNAs and circRNAs in bone marrow stem cells in osteonecrosis of the femoral head

Xiang,Shuai; Li,Zeng; Weng,Xisheng

doi:10.3892/ijmm.2020.4455

Changed cellular functions and aberrantly expressed miRNAs and circRNAs in bone marrow stem cells in osteonecrosis of the femoral head

Authors:
- Shuai Xiang
- Zeng Li
- Xisheng Weng
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Affiliations: Department of Orthopaedic Surgery, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Science, Beijing 100730, P.R. China
Published online on: January 8, 2020 https://doi.org/10.3892/ijmm.2020.4455
Pages: 805-815
Copyright: © Xiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to detect the correlations between altered cellular functions in bone marrow stem cells (BMSCs) and osteonecrosis of the femoral head (ONFH). By profiling the aberrant expression of miRNAs and circRNAs in BMSCs isolated from ONFH patients, the present study aimed to further explore the potential regulatory mechanisms of action of circRNAs in ONFH using integrated bioinformatics analysis. BMSCs were isolated from seven ONFH patients and seven controls. Cellular functions, including proliferation, apoptosis and differentiation, were compared. miRNA and circRNA sequencing were conducted using RNA samples of three ONFH patients and three controls to identify differentially expressed circRNAs and miRNAs. The expression of hsa_circ_0000219, hsa_circ_0004588 and hsa_circ_0005936 were validated by qPCR. Target miRNAs were also predicted and validated by qPCR and circRNA‑miRNA co‑expression networks were constructed. BMSCs of ONFH patients displayed decreased proliferation and increased apoptosis during in vitro culturing. In addition, reduced osteogenesis and enhanced adipogenesis were found in the ONFH group. A total of 129 miRNAs and 231 circRNAs were detected to be differentially expressed. The expression levels of hsa_circ_0000219, hsa_circ_0004588 and hsa_circ_0005936 were significantly decreased in BMSCs of ONFH patients. A number of target miRNAs related to cell proliferation, apoptosis and differentiation were predicted for hsa_circ_0000219 and hsa_circ_0005936. The expression levels of miR‑144‑3p and miR‑1270 were found to be elevated in ONFH patients, which was consistent with miRNA sequencing data and competitive endogenous RNA hypothesis. Time‑dependent expression patterns of hsa_circ_0000219, hsa_circ_0004588, hsa_circ_0005936, miR‑144‑3p and miR‑1270 were also validated during osteogenic and adipogenic differentiation in BMSCs. The results of the present study substantiated the involvement of BMSCs in ONFH development. hsa_circ_0000219 and hsa_circ_0005936 may regulate the progression of ONFH by mediating the proliferation and differentiation of BMSCs by sponging miRNAs.

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1	Cui L, Zhuang Q, Lin J, Jin J, Zhang K, Cao L, Lin J, Yan S, Guo W, He W, et al: Multicentric epidemiologic study on six thousand three hundred and ninety five cases of femoral head osteonecrosis in China. Int Orthop. 40:267–276. 2016. View Article : Google Scholar
2	Wang C, Peng J and Lu S: Summary of the various treatments for osteonecrosis of the femoral head by mechanism: A review. Exp Ther Med. 8:700–706. 2014. View Article : Google Scholar : PubMed/NCBI
3	Houdek MT, Wyles CC, Packard BD, Terzic A, Behfar A and Sierra RJ: Decreased osteogenic activity of mesenchymal stem cells in patients with corticosteroid-induced osteonecrosis of the femoral head. J Arthroplasty. 31:893–898. 2016. View Article : Google Scholar
4	Yeh CH, Chang JK, Ho ML, Chen CH and Wang GJ: Different differentiation of stroma cells from patients with osteonecrosis: A pilot study. Clin Orthop Relat Res. 467:2159–2167. 2009. View Article : Google Scholar : PubMed/NCBI
5	Balla B, Pintér C, Kósa JP, Podani J, Takács I, Nagy Z, Speer G, Horváth B, Korányi L and Lakatos P: Gene expression changes in femoral head necrosis of human bone tissue. Dis Markers. 31:25–32. 2011. View Article : Google Scholar : PubMed/NCBI
6	Wu X, Zhang Y, Guo X, Xu H, Xu Z, Duan D and Wang K: Identification of differentially expressed microRNAs involved in non-traumatic osteonecrosis through microRNA expression profiling. Gene. 565:22–29. 2015. View Article : Google Scholar : PubMed/NCBI
7	Zhao R, Li Y, Lin Z, Wan J, Xu C, Zeng Y and Zhu Y: miR-199b-5p modulates BMSC osteogenesis via suppressing GSK-3β/β-catenin signaling pathway. Biochem Biophys Res Commun. 477:749–754. 2016. View Article : Google Scholar : PubMed/NCBI
8	Li T, Li H, Wang Y, Li T, Fan J, Xiao K, Zhao RC and Weng X: microRNA-23a inhibits osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting LRP5. Int J Biochem Cell Biol. 72:55–62. 2016. View Article : Google Scholar : PubMed/NCBI
9	Salmena L, Poliseno L, Tay Y, Kats L and Pandolfi PP: A ceRNA hypothesis: The Rosetta Stone of a hidden RNA language? Cell. 146:353–358. 2011. View Article : Google Scholar : PubMed/NCBI
10	Wang Q, Li Y and Zhang Y, Ma L, Lin L, Meng J, Jiang L, Wang L, Zhou P and Zhang Y: LncRNA MEG3 inhibited osteogenic differentiation of bone marrow mesenchymal stem cells from postmenopausal osteoporosis by targeting miR-133a-3p. Biomed Pharmacother. 89:1178–1186. 2017. View Article : Google Scholar : PubMed/NCBI
11	Weng J, Peng W, Zhu S and Chen S: Long noncoding RNA sponges miR-454 to promote osteogenic differentiation in maxillary sinus membrane stem cells. Implant Dent. 26:178–186. 2017. View Article : Google Scholar : PubMed/NCBI
12	Jeck WR, Sorrentino JA, Wang K, Slevin MK, Burd CE, Liu J, Marzluff WF and Sharpless NE: Circular RNAs are abundant, conserved, and associated with ALU repeats. RNA. 19:141–157. 2013. View Article : Google Scholar :
13	Liu Q, Zhang X, Hu X, Dai L, Fu X, Zhang J and Ao Y: Circular RNA related to the chondrocyte ECM regulates MMP13 expression by functioning as a miR-136 'Sponge' in human cartilage degradation. Sci Rep. 6:225722016. View Article : Google Scholar : PubMed/NCBI
14	Dou C, Cao Z, Yang B, Ding N, Hou T, Luo F, Kang F, Li J, Yang X, Jiang H, et al: Changing expression profiles of lncRNAs, mRNAs, circRNAs and miRNAs during osteoclastogenesis. Sci Rep. 6:214992016. View Article : Google Scholar : PubMed/NCBI
15	Ouyang Q, Wu J, Jiang Z, Zhao J, Wang R, Lou A, Zhu D, Shi GP and Yang M: Microarray expression profile of circular RNAs in peripheral blood mononuclear cells from rheumatoid arthritis patients. Cell Physiol Biochem. 42:651–659. 2017. View Article : Google Scholar : PubMed/NCBI
16	Peng W, Zhu S, Chen J, Wang J, Rong Q and Chen S: Hsa_ circRNA_33287 promotes the osteogenic differentiation of maxillary sinus membrane stem cells via miR-214-3p/Runx3. Biomed Pharmacother. 109:1709–1717. 2019. View Article : Google Scholar
17	Qian DY, Yan GB, Bai B, Chen Y, Zhang SJ, Yao YC and Xia H: Differential circRNA expression profiles during the BMP2-induced osteogenic differentiation of MC3T3-E1 cells. Biomed Pharmacother. 90:492–499. 2017. View Article : Google Scholar : PubMed/NCBI
18	Otsuru S, Hofmann TJ, Olson TS, Dominici M and Horwitz EM: Improved isolation and expansion of bone marrow mesenchymal stromal cells using a novel marrow filter device. Cytotherapy. 15:146–153. 2013. View Article : Google Scholar : PubMed/NCBI
19	Gao Y, Wang J and Zhao F: CIRI: An efficient and unbiased algorithm for de novo circular RNA identification. Genome Biol. 16:42015. View Article : Google Scholar : PubMed/NCBI
20	Liu X, Shi X, Chen C and Zhang L: Improving RNA-Seq expression estimation by modeling isoform- and exon-specific read sequencing rate. BMC Bioinformatics. 16:3322015. View Article : Google Scholar : PubMed/NCBI
21	Friedländer MR, Mackowiak SD, Li N, Chen W and Rajewsky N: miRDeep2 accurately identifies known and hundreds of novel microRNA genes in seven animal clades. Nucleic Acids Res. 40:37–52. 2012. View Article : Google Scholar :
22	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
23	Gu W, Sun Y, Zheng X, Ma J, Hu XY, Gao T and Hu MJ: Identification of gastric cancer-related circular RNA through microarray analysis and bioinformatics analysis. Biomed Res Int. 2018:23816802018. View Article : Google Scholar : PubMed/NCBI
24	Song H, Tao L, Wang F, Wang W, Wei Y, Shen W and Zhou F: Effect of bone mesenchymal stem cells transplantation on the micro-environment of early osteonecrosis of the femoral head. Int J Clin Exp Pathol. 8:14528–14534. 2015.
25	Hernigou P, Beaujean F and Lambotte JC: Decrease in the mesenchymal stem-cell pool in the proximal femur in corticosteroid-induced osteonecrosis. J Bone Joint Surg Br. 81:349–355. 1999. View Article : Google Scholar : PubMed/NCBI
26	Song K, Huang M, Shi Q, Du T and Cao Y: Cultivation and identification of rat bone marrow-derived mesenchymal stem cells. Mol Med Rep. 10:755–760. 2014. View Article : Google Scholar : PubMed/NCBI
27	Yue J, Wan F, Zhang Q, Wen P, Cheng L, Li P and Guo W: Effect of glucocorticoids on miRNA expression spectrum of rat femoral head microcirculation endothelial cells. Gene. 651:126–133. 2018. View Article : Google Scholar : PubMed/NCBI
28	Han B, Chao J and Yao H: Circular RNA and its mechanisms in disease: From the bench to the clinic. Pharmacol Ther. 187:31–44. 2018. View Article : Google Scholar : PubMed/NCBI
29	Qu S, Yang X, Li X, Wang J, Gao Y, Shang R, Sun W, Dou K and Li H: Circular RNA: A new star of noncoding RNAs. Cancer Lett. 365:141–148. 2015. View Article : Google Scholar : PubMed/NCBI
30	Gu X, Li M, Jin Y, Liu D and Wei F: Identification and integrated analysis of differentially expressed lncRNAs and circRNAs reveal the potential ceRNA networks during PDLSC osteogenic differentiation. BMC Genet. 18:1002017. View Article : Google Scholar : PubMed/NCBI
31	Liao Q, Liu C, Yuan X, Kang S, Miao R, Xiao H, Zhao G, Luo H, Bu D, Zhao H, et al: Large-scale prediction of long non-coding RNA functions in a coding-non-coding gene co-expression network. Nucleic Acids Res. 39:3864–3878. 2011. View Article : Google Scholar : PubMed/NCBI
32	De Braekeleer E, Auffret R, Douet-Guilbert N, Basinko A, Le Bris MJ, Morel F and De Braekeleer M: Recurrent translocation (10;17)(p15;q21) in acute poorly differentiated myeloid leukemia likely results in ZMYND11-MBTD1 fusion. Leuk Lymphoma. 55:1189–1190. 2014. View Article : Google Scholar
33	Qi F, Hu JF, Liu BH, Wu CQ, Yu HY, Yao DK and Zhu L: miR-9a-5p regulates proliferation and migration of hepatic stellate cells under pressure through inhibition of Sirt1. World J Gastroenterol. 21:9900–9915. 2015. View Article : Google Scholar : PubMed/NCBI
34	Lin CH, Li NT, Cheng HS and Yen ML: Oxidative stress induces imbalance of adipogenic/osteoblastic lineage commitment in mesenchymal stem cells through decreasing SIRT1 functions. J Cell Mol Med. 22:786–796. 2018.
35	Yuan HF, Zhai C, Yan XL, Zhao DD, Wang JX, Zeng Q, Chen L, Nan X, He LJ, Li ST, et al: SIRT1 is required for long-term growth of human mesenchymal stem cells. J Mol Med (Berl). 90:389–400. 2012. View Article : Google Scholar
36	Chen JS, Li HS, Huang JQ, Dong SH, Huang ZJ, Yi W, Zhan GF, Feng JT, Sun JC and Huang XH: MicroRNA-379-5p inhibits tumor invasion and metastasis by targeting FAK/AKT signaling in hepatocellular carcinoma. Cancer Lett. 375:73–83. 2016. View Article : Google Scholar : PubMed/NCBI
37	Sun C, Yuan H, Wang L, Wei X, Williams L, Krebsbach PH, Guan JL and Liu F: FAK promotes osteoblast progenitor cell proliferation and differentiation by enhancing Wnt signaling. J Bone Miner Res. 31:2227–2238. 2016. View Article : Google Scholar : PubMed/NCBI
38	Xu Y, Zhang G, Zou C, Zhang H, Gong Z, Wang W, Ma G, Jiang P and Zhang W: LncRNA MT1JP suppresses gastric cancer cell proliferation and migration through MT1JP/miR-214-3p/RUNX3 axis. Cell Physiol Biochem. 46:2445–2459. 2018. View Article : Google Scholar : PubMed/NCBI
39	Wang Y, Feng Q, Ji C, Liu X, Li L and Luo J: RUNX3 plays an important role in mediating the BMP9-induced osteogenic differentiation of mesenchymal stem cells. Int J Mol Med. 40:1991–1999. 2017.PubMed/NCBI
40	Huang C, Geng J, Wei X, Zhang R and Jiang S: miR-144-3p regulates osteogenic differentiation and proliferation of murine mesenchymal stem cells by specifically targeting Smad4. FEBS Lett. 590:795–807. 2016. View Article : Google Scholar : PubMed/NCBI
41	Gao W, Wu Y, He X, Zhang C, Zhu M, Chen B, Liu Q, Qu X, Li W, Wen S and Wang B: MicroRNA-204-5p inhibits invasion and metastasis of laryngeal squamous cell carcinoma by suppressing forkhead box C1. J Cancer. 8:2356–2368. 2017. View Article : Google Scholar : PubMed/NCBI
42	Hopkins A, Mirzayans F and Berry F: Foxc1 expression in early osteogenic differentiation is regulated by BMP4-SMAD activity. J Cell Biochem. 117:1707–1717. 2016. View Article : Google Scholar
43	Gu Y, Shi Y, Yang Q, Gu WW, He YP, Zheng HJ, Zhang X, Wang JM and Wang J: miR-3074-5p promotes the apoptosis but inhibits the invasiveness of human extravillous trophoblast-derived HTR8/SVneo cells in vitro. Reprod Sci. 25:690–699. 2018. View Article : Google Scholar
44	Li Z, Jiang C, Li X, Wu WKK, Chen X, Zhu S, Ye C, Chan MTV and Qian W: Circulating microRNA signature of steroid-induced osteonecrosis of the femoral head. Cell Prolif. 51:2018. View Article : Google Scholar
45	Zhang MY, Wang JB, Zhu ZW, Li LJ, Liu RS, Yang XK, Leng RX, Li XM, Pan HF and Ye DQ: Differentially expressed circular RNAs in systemic lupus erythematosus and their clinical significance. Biomed Pharmacother. 107:1720–1727. 2018. View Article : Google Scholar : PubMed/NCBI
46	Li LJ, Zhu ZW, Zhao W, Tao SS, Li BZ, Xu SZ, Wang JB, Zhang MY, Wu J, Leng RX, et al: Circular RNA expression profile and potential function of hsa_circ_0045272 in systemic lupus erythematosus. Immunology. 155:137–149. 2018. View Article : Google Scholar : PubMed/NCBI