[1]张建军△,谭章琴,冯璐.环状核糖核酸0136474调控骨关节炎软骨细胞增殖和凋亡的分子机制研究[J].中国中医骨伤科杂志,2024,32(03):23-29.[doi:10.20085/j.cnki.issn1005-0205.240305]
　ZHANG Jianjun,TAN Zhangqin,FENG Lu.Molecular Mechanism Study of Circular Ribonucleic Acid 0136474 in Regulating Proliferation and Apoptosis of Osteoarthritis Chondrocytes[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2024,32(03):23-29.[doi:10.20085/j.cnki.issn1005-0205.240305]
点击复制

环状核糖核酸0136474调控骨关节炎软骨细胞增殖和凋亡的分子机制研究()
分享到：

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第32卷
期数:
2024年03期
页码:
23-29
栏目:
实验研究
出版日期:
2024-03-15

文章信息/Info

Title:
Molecular Mechanism Study of Circular Ribonucleic Acid 0136474 in Regulating Proliferation and Apoptosis of Osteoarthritis Chondrocytes
文章编号:
1005-0205(2024)03-0023-07
作者:
张建军1△; 谭章琴1; 冯璐1
1简阳市中医医院(四川 简阳,641400)
Author(s):
ZHANG Jianjun1△; TAN Zhangqin1; FENG Lu1
1Jianyang Chinese Medicine Hospital,Jianyang 641400,Sichuan China.
关键词:
环状核糖核酸;  微小核糖核酸;  骨关节炎;  软骨细胞增殖;  软骨细胞凋亡
Keywords:
circular ribonucleic acid;  micro ribonucleic acid;  osteoarthritis;  chondrocyte proliferation;  chondrocyte apoptosis
分类号:
R684
DOI:
10.20085/j.cnki.issn1005-0205.240305
文献标志码:
A
摘要:
目的:探讨环状RNA-0136474(circ_0136474)调控微小RNA-127-5p(miR-127-5p)/Zest 同源物2(EZH2)轴对骨关节炎(OA)软骨细胞增殖和凋亡的影响及其分子机制。方法:收集骨关节炎患者软骨组织并检测其中circ_0136474表达; IL-1β诱导人软骨细胞CHON-001,并分为对照组、IL-1β组、si-NC组、si-circ_0136474组、si-circ_0136474+anti-miR-NC组、si-circ_0136474+anti-miR-127-5p组、miR-NC组和miR-127-5p mimics组,CCK-8法检测细胞活力,流式细胞仪检测细胞周期和细胞凋亡,RT-qPCR检测circ_0136474、miR-127-5p及EZH2、增殖细胞核抗原(PCNA)mRNA表达,ELISA试剂盒检测IL-6、TNF-α、MMP-13含量,Western Blot检测增殖细胞核抗原、Bcl-2、Bax、EZH2蛋白表达,双荧光素酶报告基因实验验证miR-127-5p与circ_0136474、EZH2的靶向关系。结果:circ_0136474在骨关节炎患者软骨组织中高表达。经IL-1β诱导后,细胞活力、S期、PCNA表达、Bcl-2表达和miR-127-5p显著降低,G0～G1期细胞比例、细胞凋亡率、Bax、IL-6、TNF-α、MMP-13、circ_0136474、EZH2蛋白和EZH2 mRNA显著增加(P<0.05); 双荧光素酶报告基因实验显示,circ_0136474、EZH2与miR-127-5p存在靶向关系,沉默circ_0136474可促进细胞增殖、延长S期,缩短G0～G1期,并抑制细胞凋亡,减少IL-6、TNF-α、MMP-13含量(P<0.05); 抑制miR-127-5p表达可逆转沉默circ_0136474对细胞凋亡、IL-6、TNF-α、MMP-13的抑制作用(P<0.05)。结论:circ_0136474在骨关节炎软骨组织中高表达,沉默circ_0136474可通过调节miR-127-5p/EZH2轴促进软骨细胞增殖而抑制细胞凋亡。
Abstract:
Objective:To investigate the effect of circular RNA-0136474(circ_0136474)on the proliferation and apoptosis of osteoarthritis chondrocytes by regulating the micro RNA-127-5p(miR-127-5p)/Zest homolog 2(EZH2)axis and its molecular mechanism.Methods:The osteoarthritis patients were collected.The expression of circ_0136474 in cartilage tissue of osteoarthritis patients was detected; human chondrocytes CHON-001 were induced by IL-1β and divided into control group,IL-1β group,si-NC group,si-circ_0136474 group,si-circ_0136474+anti-miR-NC group,si-circ_0136474+anti-miR-127-5p group,miR-NC group,and miR-127-5p mimics group.The CCK-8 was applied to detect the cell viability,flow cytometer was applied to detect the cell cycle and apoptosis,RT-qPCR was applied to detect the expression of circ_0136474,miR-127-5p,EZH2,and proliferating cell nuclear antigen(PCNA)mRNA,ELISA kit was applied to detect the levels of IL-6,TNF-α,and MMP-13,Western Blot was applied to detect the expression of PCNA,Bcl-2,Bax,and EZH2 proteins,double luciferase reporter gene experiment was applied to verify the targeting relationship between miR-127-5p and circ_0136474,EZH2.Results:The circ_0136474 was highly expressed in cartilage tissue of osteoarthritis patients; after induction by IL-1β,the cell viability,S phase,PCNA expression,Bcl-2 expression,and miR-127-5p were obviously reduced,the proportion of G0-G1 phase cells,cell apoptosis rate,Bax,IL-6,TNF-α,MMP-13,circ_0136474,EZH2 protein,and EZH2 mRNA were obviously increased(P<0.05).The double luciferase reporter gene experiment showed that circ_0136474 and EZH2 had a targeting relationship with miR-127-5p,silencing circ_0136474 can promote cell proliferation,prolong the S phase,shorten the G0-G1 phase,inhibit cell apoptosis,reduce the levels of IL-6,TNF-α,and MMP-13(P<0.05),inhibiting the expression of miR-127-5p can reverse the inhibitory effects of silencing circ_0136474 on cell apoptosis,IL-6,TNF-α,and MMP-13(P<0.05).Conclusion:The circ_0136474 is highly expressed in osteoarthritis cartilage tissue,and silencing circ_0136474 can promote chondrocyte proliferation and inhibit cell apoptosis by regulating the miR-127-5p/EZH2 axis.

参考文献/References:

[1] EMAMI A,NAMDARI H,PARVIZPOUR F,et al.Challenges in osteoarthritis treatment[J].Tissue Cell,2023,80(1):101992-102003.
[2] HE Y,LI Z,ALEXANDER P G,et al.Pathogenesis of osteoarthritis:risk factors,regulatory pathways in chondrocytes,and experimental models[J].Biology(Basel),2020,9(8):194-227.
[3] CHENG S,NIE Z,CAO J,et al.Circ_0136474 promotes the progression of osteoarthritis by sponging miR-140-3p and upregulating MECP2[J].J Mol Histol,2023,54(1):1-12.
[4] LI Z,YUAN B,PEI Z,et al.Circ_0136474 and MMP-13 suppressed cell proliferation by competitive binding to miR-127-5p in osteoarthritis[J].J Cell Mol Med,2019,23(10):6554-6564.
[5] LUO L,WANG Z,HU T,et al.Multiomics characteristics and immunotherapeutic potential of EZH2 in pan-cancer[J].Biosci Rep,2023,43(1):BSR20222230-BSR20222252.
[6] LUI JC,GARRISON P,NGUYEN Q,et al.EZH1 and EZH2 promote skeletal growth by repressing inhibitors of chondrocyte proliferation and hypertrophy[J].Nat Commun,2016,7(1):13685-13696.
[7] ALLAS L,BROCHARD S,ROCHOUX Q,et al.EZH2 inhibition reduces cartilage loss and functional impairment related to osteoarthritis[J].Sci Rep,2020,10(1):19577-19589.
[8] CHEN L,WU Y,WU Y,et al.The inhibition of EZH2 ameliorates osteoarthritis development through the Wnt/β-catenin pathway[J].Sci Rep,2016,6(1):29176-29189.
[9] ZHU H,ZHU S,SHANG X,et al.Exhausting circ_0136474 and restoring miR-766-3p attenuate chondrocyte oxidative injury in IL-1β-induced osteoarthritis progression through regulating DNMT3A[J].Front Genet,2021,12(1):1-13.
[10] LI Z,LU J.CircRNAs in osteoarthritis:research status and prospect[J].Front Genet,2023,14(1):1-19.
[11] ZHANG Y,LIU L,LIU K,et al.Regulatory mechanism of circular RNA involvement in osteoarthritis[J].Front Surg,2022,9(1):1-13.
[12] PAN F,LI Z,LUO Y,et al.Circ_0136474 contributes to the IL-1β-induced chondrocyte injury by binding to miR-665 to induce the FGFR1 upregulation[J].Transpl Immunol,2022:101615.
[13] LIU H,ZHAO H,HUANG Y,et al.Circ_0002715 promotes the development of osteoarthritis through regulating LXN by sponging miR-127-5p[J].J Orthop Surg Res,2023,18(1):230-240.
[14] ZHANG Y,ZHAO P,LI S,et al.CircSCAPER knockdown attenuates IL-1β-induced chondrocyte injury by miR-127-5p/TLR4 axis in osteoarthritis[J].Autoimmunity,2022,55(8):577-586.
[15] LIU C,CHENG P,LIANG J,et al.Circular RNA circ_0128846 promotes the progression of osteoarthritis by regulating miR-127-5p/NAMPT axis[J].J Orthop Surg Res,2021,16(1):307-318.
[16] HWANG I J,PARK J,SEO S B.Non-canonical transcriptional regulation of INHAT subunit SET/TAF-Iβ by EZH2[J].Biochem Biophys Res Commun,2022,635(1):136-143.
[17] DU X,CHEN Y,ZHANG Q,et al.EZH2 ameliorates osteoarthritis by activating TNFSF13B[J].J Bone Miner Res,2020,35(5):956-965.
[18] LUOBU Z,WANG L,JIANG D,et al.CircSCAPER contributes to IL-1β-induced osteoarthritis in vitro via miR-140-3p/EZH2 axis[J].Bone Joint Res,2022,11(2):61-72.
[19] LI Y,YUAN F,SONG Y,et al.miR-17-5p and miR-19b-3p prevent osteoarthritis progression by targeting EZH2[J].Exp Ther Med,2020,20(2):1653-1663.
[20] CHENG F,LI H,LIU J,et al.EZH2 regulates the balance between osteoclast and osteoblast differentiation to inhibit arthritis-induced bone destruction[J].Genes Immun,2022,23(3/4):141-148.

备注/Memo

备注/Memo:
基金项目:四川省中医药信息学会关于舒适化医疗(新晨基金)专项科研课题(20200119)
△通信作者 E-mail:znmc11@163.com
更新日期/Last Update: 2024-03-15