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Published on: June 16, 2023
Knockdown of circ-PRKCH alleviates IL-1β-treated chondrocyte cell phenotypic changes through modulating
Zhongxing Liu1, Jian Cao1, Limin Zhang1
1Department of Orthopedics, Affiliated Hospital of Chifeng University, Institute of Orthopaedic Diseases, Affiliated Hospital of Chifeng University, Chifeng, China.
Background:
Osteoarthritis (OA) is a common joint disease characterized by progressive cartilage degradation. Circular RNAs (circRNAs) are involved in the initiation and development of OA. This study aimed to explore the potential role and mechanism of circRNA protein kinase C eta (circ-PRKCH) in OA.
Methods:
A total of 30 cartilage specimens were collected from OA patients or normal subjects. Human chondrocytes (CHON-001) were stimulated with interleukin-1β (IL-1β) to establish an in vitro OA model. The expression levels of circ-PRKCH, microRNA-502-5p (miR-502-5p) and circ-PRKCH or A disintegrin and metalloproteases metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) in cartilage specimens and IL-1β-treated chondrocytes were detected by quantitative real-time PCR or Western blot, and their correlation in OA cartilage specimens was analysed by Spearman's correlation coefficient. The targeted relationship between miR-502-5p and circ-PRKCH or ADAMTS5 was verified by dual-luciferase reporter assay and RNA Immunoprecipitation (RIP) assay. Cell Counting Kit-8 (CCK-8), 5-Ethynyl-2'-deoxyuridine (EDU), flow cytometry, wound healing and enzyme-linked immunosorbent assay (ELISA) assays were applied to evaluate cell proliferation, apoptosis, migration and inflammatory response in IL-1β-treated chondrocytes. Exosomes were identified by transmission electron microscope (TEM) and Western blot.
Results:
Circ-PRKCH and ADAMTS5 expression levels were up-regulated, while miR-502-5p expression was down-regulated in OA cartilage tissues and IL-1β-treated chondrocytes. Depletion of circ-PRKCH relieved IL-1β-treated chondrocyte cell phenotypic changes by promoting cell proliferation and migration, as well as inhibiting apoptosis and inflammatory response. Mechanically, circ-PRKCH acted as a sponge for miR-502-5p to regulate ADAMTS5 expression, thereby contributing to IL-1β-treated chondrocyte cell phenotypic changes. Moreover, exosomes derived from IL-1β-treated chondrocytes could transfer circ-PRKCH across cells.
Conclusion:
Circ-PRKCH contributed to IL-1β-treated cell phenotypic changes in chondrocytes via modulating miR-502-5p/ADAMTS5 pathway, which might provide a promising biomarker for OA treatment.
Insights
Circular RNA protein kinase C eta (circ-PRKCH) exacerbates osteoarthritis by sponging miR-502-5p and upregulating ADAMTS5. This circRNA may serve as a therapeutic target for osteoarthritis, offering a novel approach to cartilage degradation treatment.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Osteoarthritis (OA) is a prevalent degenerative joint disease.
- Circular RNAs (circRNAs) play a role in OA pathogenesis.
- The specific role of circRNA protein kinase C eta (circ-PRKCH) in OA requires further investigation.
Purpose of the Study:
- To investigate the role and mechanism of circ-PRKCH in osteoarthritis.
- To explore the regulatory relationship between circ-PRKCH, miR-502-5p, and ADAMTS5 in OA.
- To assess the potential of circ-PRKCH as a biomarker for OA treatment.
Main Methods:
- Quantitative real-time PCR and Western blot were used to measure gene and protein expression.
- Dual-luciferase reporter and RNA Immunoprecipitation (RIP) assays verified molecular interactions.
- Cellular assays (CCK-8, EDU, flow cytometry, ELISA) assessed chondrocyte function.
- Exosome identification was performed using TEM and Western blot.
Main Results:
- Circ-PRKCH and ADAMTS5 were upregulated, while miR-502-5p was downregulated in OA cartilage and chondrocytes.
- Circ-PRKCH depletion improved chondrocyte viability, proliferation, and migration while reducing apoptosis and inflammation.
- Circ-PRKCH acts as a sponge for miR-502-5p, regulating ADAMTS5 expression.
- Exosomes from OA chondrocytes can transfer circ-PRKCH.
Conclusions:
- Circ-PRKCH promotes OA progression by modulating the miR-502-5p/ADAMTS5 pathway in chondrocytes.
- Circ-PRKCH may serve as a potential therapeutic target and biomarker for osteoarthritis.
