Long non-coding RNA MCM3AP-AS1 protects chondrocytes ATDC5 and CHON-001 from IL-1β-induced inflammation via regulating miR-138-5p/SIRT1

• Published in: Bioengineered Vol. 12; no. 1; pp. 1445 - 1456
• Main Authors: Shi, Jianming, Cao, Fuyang, Chang, Yingjian, Xin, Chaofei, Jiang, Xu, Xu, Jianzhong, Lu, Shitao
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis 01.01.2021
• Subjects: Cell Line; Chondrocytes - drug effects; Chondrocytes - metabolism; Female; Humans; Inflammation - metabolism; Interleukin-1beta - genetics; Interleukin-1beta - metabolism; Male; MCM3AP-as1; MicroRNAs - genetics; MicroRNAs - metabolism; Middle Aged; miR-138-5p; osteoarthritis; Osteoarthritis - metabolism; Research Paper; RNA, Long Noncoding - genetics; RNA, Long Noncoding - metabolism; RNA, Long Noncoding - pharmacology; sirt1; Sirtuin 1 - genetics; Sirtuin 1 - metabolism; miR-138-5p; MCM3AP-as1; osteoarthritis; sirt1
• ISSN: 2165-5979; 2165-5987
• DOI: 10.1080/21655979.2021.1905247

Osteoarthritis (OA) is a chronic inflammatory joint disease. Increased apoptosis of chondrocytes contributes to cartilage degradation in OA pathogenesis. The function of lncRNA MCM3AP-AS1 in regulating the viability of chondrocytes still awaits further elaboration. In this work, MCM3AP-AS1, miR-138-5p and SIRT1 mRNA expression levels in OA and normal cartilage tissues were detected by qRT-PCR. Besides, chondrocyte cell lines, CHON-001 and ATDC5 induced by interleukin-1β (IL-1β) were used to initiate the inflammatory response environment of OA. CCK-8 assay was used to examine the cell multiplication; meanwhile, transwell assay was utilized to detect migration. Western blot was adopted to determine SIRT1 expression in chondrocyte. Enzyme-linked immunosorbent assay (ELISA) was performed to evaluate inflammatory factor levels. In addition, the binding sites between MCM3AP-AS1 and miR-138-5p, miR-138-5p and 3ʹUTR of SIRT1 were validated by dual-luciferase reporter assay, RIP assay or RNA pull-down assay. It was found that MCM3AP-AS1 was declined in OA cartilage tissues, positively interrelated with SIRT1 expression while negatively correlated with miR-138-5p. MCM3AP-AS1 up-regulation enhanced the viability and migration of CHON-001 and ATDC5 cells while restraining the apoptosis and inflammatory response. Additionally, miR-138-5p overexpression counteracted the effects on chondrocytes caused by MCM3AP-AS1 overexpression. MCM3AP-AS1 could adsorb miR-138-5p, and SIRT1 was verified as a target of miR-138-5p, and SIRT1 could be up-regulated by overexpression of MCM3AP-AS1 indirectly. In conclusion, MCM3AP-AS1 has the potential to be the 'ceRNA' to regulate miR-138-5p and SIRT1 in chondrocytes, and to participate in the pathogenesis of OA.