Research Paper Volume 12, Issue 4 pp 3218—3237

LINC00623/miR-101/HRAS axis modulates IL-1β-mediated ECM degradation, apoptosis and senescence of osteoarthritis chondrocytes

Figure 3. LINC00623 upregulates HRAS to suppress IL-1β-induced chondrocyte apoptosis and senescence. (A) LINC00623 expression in normal, mild OA and severe OA tissues was examined using real-time PCR assays. (B) OA chondrocytes were treated with PBS or IL-1βand then examined for LINC00623 expression using real-time PCR. (C) LINC00623 knockdown was achieved by transfection of sh1-LINC00623 or sh2-LINC00623, as confirmed using real-time PCR. (D) OA chondrocytes were cotransfected with sh1/2-LINC00623 and the HRAS vector in the presence or absence of IL-1β treatment and the protein levels of HRAS. (E, F) The cell apoptosis was examined using TUNEL and flow cytometer assays. (G) The SA-β-Gal positive cells were determined by the SA-β-Gal staining. (H) The protein levels of Caspase 3, cleaved-Caspase 3, Caspase 7 and cleaved-Caspase 7 were examined using immunoblotting. The data are presented as mean ± SD of three independent experiments. *P<0.05, **P<0.01, compared to control group; #P<0.05, ##P<0.01, compared to sh-LINC00623 group.