Silencing of long‑chain non‑coding RNA GAS5 in osteoarthritic chondrocytes is mediated by targeting the miR‑34a/Bcl‑2 axis

Ji,Qinghui; Qiao,Xiaofeng; Liu,Yongxiang; Wang,Dawei; Yan,Jinglong

doi:10.3892/mmr.2019.10900

Silencing of long‑chain non‑coding RNA GAS5 in osteoarthritic chondrocytes is mediated by targeting the miR‑34a/Bcl‑2 axis

Authors:
- Qinghui Ji
- Xiaofeng Qiao
- Yongxiang Liu
- Dawei Wang
- Jinglong Yan
View Affiliations

Affiliations: Department of Orthopedics, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, Department of Orthopedics, The First Affiliated Hospital of Jiamusi University, Jiamusi, Heilongjiang 154000, P.R. China, Department of Orthopedics, Hegang People's Hospital, Hegang, Heilongjiang 154100, P.R. China, Department of Orthopedics, First Hospital of Zhangjiakou, Zhangjiakou, Hebei 075000, P.R. China
Published online on: December 20, 2019 https://doi.org/10.3892/mmr.2019.10900
Pages: 1310-1319

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 investigate the effects of the long non‑coding RNA (lncRNA) growth arrest‑specific transcript 5 (GAS5) on proliferation, apoptosis and the inflammatory response of osteoarthritic chondrocytes (OACs) and its associated mechanism of action. Primary chondrocytes were isolated from cartilage tissues of osteoarthritis (OA) patients for subculture. GAS5 was silenced in OACs by liposome transfection. The effects of GAS5 silencing on proliferation, apoptosis, stromal metabolism and inflammatory response of OACs were analyzed. The association of GAS5 with its target microRNA‑34a (miR‑34a) and the downstream target gene Bcl‑2 was verified by luciferase reporter assays. The results indicated that GAS5 silencing promoted the proliferation, inhibited cell apoptosis and caused G1 arrest of OACs compared with the control group (P<0.05). The expression levels of TNF‑α and IL‑6 in the supernatant of OACs in the si‑GAS5 group were significantly lower than those of the control group (P<0.05). The results of the double luciferase reporter assays indicated that overexpression of GAS5 downregulated miR‑34a and upregulated Bcl‑2 levels (P<0.05) compared with the expression levels of these markers in the control group. In contrast to GAS5 overexpression, knockdown of this RNA caused a significant upregulation of miR‑34a levels and a significant downregulation in the levels of Bcl‑2 (P<0.05). Moreover, GAS5 overexpression could counteract the inhibition of apoptosis by overexpression of miR‑34a (P<0.05). The data indicated that GAS5 participated in the development of OA by regulating the biological behavior of chondrocytes via the miR‑34a/Bcl‑2 axis.

View Figures

View References

1	Mandl LA: Osteoarthritis year in review 2018: Clinical. Osteoarthritis Cartilage. 27:359–364. 2019. View Article : Google Scholar : PubMed/NCBI
2	Zhang C, Wang P, Jiang P, Lv Y, Dong C, Dai X, Tan L and Wang Z: Upregulation of lncRNA HOTAIR contributes to IL-1β-induced MMP overexpression and chondrocytes apoptosis in temporomandibular joint osteoarthritis. Gene. 586:248–253. 2016. View Article : Google Scholar : PubMed/NCBI
3	Xing D, Liang JQ, Li Y, Lu J, Jia HB, Xu LY and Ma XL: Identification of long noncoding RNA associated with osteoarthritis in humans. Orthop Surg. 6:288–293. 2014. View Article : Google Scholar : PubMed/NCBI
4	Liu Q, Zhang X, Dai L, Hu X, Zhu J, Li L, Zhou C and Ao Y: Long noncoding RNA related to cartilage injury promotes chondrocyte extracellular matrix degradation in osteoarthritis. Arthritis Rheumatol. 66:969–978. 2014. View Article : Google Scholar : PubMed/NCBI
5	Kino T, Hurt DE, Ichijo T, Nader N and Chrousos GP: Noncoding RNA gas5 is a growth arrest- and starvation-associated repressor of the glucocorticoid receptor. Sci Signal. 3:ra82010. View Article : Google Scholar : PubMed/NCBI
6	Hu L, Ye H, Huang G, Luo F, Liu Y, Liu Y, Yang X, Shen J, Liu Q and Zhang J: Long noncoding RNA GAS5 suppresses the migration and invasion of hepatocellular carcinoma cells via miR-21. Tumour Biol. 37:2691–2702. 2016. View Article : Google Scholar : PubMed/NCBI
7	Yao T, Lu R, Zhang J, Fang X, Fan L, Huang C, Lin R and Lin Z: Growth arrest-specific 5 attenuates cisplatin-induced apoptosis in cervical cancer by regulating STAT3 signaling via miR-21. J Cell Physiol. 234:9605–9615. 2019. View Article : Google Scholar : PubMed/NCBI
8	Song J, Ahn C, Chun CH and Jin EJ: A long non-coding RNA, GAS5, plays a critical role in the regulation of miR-21 during osteoarthritis. J Orthop Res. 32:1628–1635. 2014. View Article : Google Scholar : PubMed/NCBI
9	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
10	Chen YY, Chen Y, Wang WC, Tang Q, Wu R, Zhu WH, Li D and Liao LL: Cyclin D1 regulates osteoarthritis chondrocyte apoptosis via WNT3/β-catenin signalling. Artif Cells Nanomed Biotechnol. 47:1971–1977. 2019. View Article : Google Scholar : PubMed/NCBI
11	Vedicherla S and Buckley CT: In vitro extracellular matrix accumulation of nasal and articular chondrocytes for intervertebral disc repair. Tissue Cell. 49:503–513. 2017. View Article : Google Scholar : PubMed/NCBI
12	Vila S: Inflammation in Osteoarthritis. P R Health Sci J. 36:123–129. 2017.PubMed/NCBI
13	Niu J, Clancy M, Aliabadi P, Vasan R and Felson DT: Metabolic syndrome, its components, and knee osteoarthritis: The framingham osteoarthritis study. Arthritis Rheumatol. 69:1194–1203. 2017. View Article : Google Scholar : PubMed/NCBI
14	Hayashi S, Nishiyama T, Miura Y, Fujishiro T, Kanzaki N, Hashimoto S, Matsumoto T, Kurosaka M and Kuroda R: DcR3 induces cell proliferation through MAPK signaling in chondrocytes of osteoarthritis. Osteoarthritis Cartilage. 19:903–910. 2011. View Article : Google Scholar : PubMed/NCBI
15	Almonte-Becerril M, Navarro-Garcia F, Gonzalez-Robles A, Vega-Lopez MA, Lavalle C and Kouri JB: Cell death of chondrocytes is a combination between apoptosis and autophagy during the pathogenesis of Osteoarthritis within an experimental model. Apoptosis. 15:631–638. 2010. View Article : Google Scholar : PubMed/NCBI
16	Blanco FJ, Guitian R, Vazquez-Martul E, de Toro FJ and Galdo F: Osteoarthritis chondrocytes die by apoptosis. A possible pathway for osteoarthritis pathology. Arthritis Rheum. 41:284–289. 1998. View Article : Google Scholar : PubMed/NCBI
17	Li Y, Li S, Luo Y, Liu Y and Yu N: LncRNA PVT1 regulates chondrocyte apoptosis in osteoarthritis by acting as a sponge for miR-488-3p. DNA Cell Biol. 36:571–580. 2017. View Article : Google Scholar : PubMed/NCBI
18	Zhang G, Wu Y, Xu D and Yan X: Long noncoding RNA UFC1 promotes proliferation of chondrocyte in osteoarthritis by acting as a sponge for miR-34a. DNa Cell Biol. 35:691–695. 2016. View Article : Google Scholar : PubMed/NCBI
19	Chen L, Yang W, Guo Y, Chen W, Zheng P, Zeng J and Tong W: Exosomal lncRNA GAS5 regulates the apoptosis of macrophages and vascular endothelial cells in atherosclerosis. PLoS One. 12:e1854062017.
20	Mourtada-Maarabouni M and Williams GT: Role of GAS5 noncoding RNA in mediating the effects of rapamycin and its analogues on mantle cell lymphoma cells. Clin Lymphoma Myeloma Leuk. 14:468–473. 2014. View Article : Google Scholar : PubMed/NCBI
21	Shi X, Sun M, Liu H, Yao Y, Kong R, Chen F and Song Y: A critical role for the long non-coding RNA GAS5 in proliferation and apoptosis in non-small-cell lung cancer. Mol Carcinog. 54 (Suppl 1):E1–E12. 2015. View Article : Google Scholar : PubMed/NCBI
22	Udayakumar T, Shareef MM, Diaz DA, Ahmed MM and Pollack A: The E2F1/Rb and p53/MDM2 pathways in DNA repair and apoptosis: Understanding the crosstalk to develop novel strategies for prostate cancer radiotherapy. Semin Radiat Oncol. 20:258–266. 2010. View Article : Google Scholar : PubMed/NCBI
23	Onuora S: Osteoarthritis: Cartilage matrix stiffness regulates chondrocyte metabolism and OA pathogenesis. Nat Rev Rheumatol. 11:5042015. View Article : Google Scholar
24	Tang LP, Ding JB, Liu ZH and Zhou GJ: LncRNA TUG1 promotes osteoarthritis-induced degradation of chondrocyte extracellular matrix via miR-195/MMP-13 axis. Eur Rev Med Pharmacol Sci. 22:8574–8581. 2018.PubMed/NCBI
25	Li YF, Li SH, Liu Y and Luo YT: Long noncoding RNA CIR promotes chondrocyte extracellular matrix degradation in osteoarthritis by acting as a sponge for Mir-27b. Cell Physiol Biochem. 43:602–610. 2017. View Article : Google Scholar : PubMed/NCBI
26	Liu Q, Hu X, Zhang X, Dai L, Duan X, Zhou C and Ao Y: The TMSB4 pseudogene lncRNA functions as a competing endogenous RNA to promote cartilage degradation in human osteoarthritis. Mol Ther. 24:1726–1733. 2016. View Article : Google Scholar : PubMed/NCBI
27	Nixon AJ, Grol MW, Lang HM, Ruan MZC, Stone A, Begum L, Chen Y, Dawson B, Gannon F, Plutizki S, et al: Disease-modifying osteoarthritis treatment with interleukin-1 receptor antagonist gene therapy in small and large animal models. Arthritis Rheumatol. 70:1757–1768. 2018. View Article : Google Scholar : PubMed/NCBI
28	Kumar MM and Goyal R: LncRNA as a therapeutic target for angiogenesis. Curr Top Med Chem. 17:1750–1757. 2017. View Article : Google Scholar : PubMed/NCBI
29	Tong X, Gu PC, Xu SZ and Lin XJ: Long non-coding RNA-DANCR in human circulating monocytes: A potential biomarker associated with postmenopausal osteoporosis. Biosci Biotechnol Biochem. 79:732–737. 2015. View Article : Google Scholar : PubMed/NCBI
30	Yan X, Tang B, Chen B, Shan Y, Yang H; Reproducibility Project: Cancer Biology, ; Iorns E, Tsui R, Denis A, Perfito N and Errington TM: Replication Study: The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Elife. 8(pii): e435112019. View Article : Google Scholar : PubMed/NCBI
31	He R, Liu P, Xie X, Zhou Y, Liao Q, Xiong W, Li X, Li G, Zeng Z and Tang H: circGFRA1 and GFRA1 act as ceRNAs in triple negative breast cancer by regulating miR-34a. J Exp Clin Cancer Res. 36:1452017. View Article : Google Scholar : PubMed/NCBI
32	Sun TY, Xie HJ, Li Z, Kong LF, Gou XN, Li DJ, Shi YJ and Ding YZ: miR-34a regulates HDAC1 expression to affect the proliferation and apoptosis of hepatocellular carcinoma. Am J Transl Res. 9:103–114. 2017.PubMed/NCBI
33	Huang Q, Zheng Y, Ou Y, Xiong H, Yang H, Zhang Z, Chen S and Ye Y: miR-34a/Bcl-2 signaling pathway contributes to age-related hearing loss by modulating hair cell apoptosis. Neurosci Lett. 661:51–56. 2017. View Article : Google Scholar : PubMed/NCBI