LINC00052靶向miR-145发挥对TNF-α诱导人关节软骨细胞损伤的保护作用

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Abstract

Objective To identify the role of long non-coding RNA LINC00052 in the injury of tumor necrosis factor alpha(TNF-α)-induced human articular chondrocyte，and to elucidate its regulatory mechanism on miR-145.Methods The injury model of human C-20/A4 articular chondrocyte was induced and constructed by using 0，10 and 30 ng/mL of TNF-α.Real-time quantitative polymerase chain reaction(RT-qPCR) was used to detect the expression levels of LINC00052 and miR-145 and their expression correlation.The specific short hairpin RNA was used to interfere with the expression of LINC00052 in C-20/A4 cells，and cell proliferation ability was detected by 3-(4，5-Dimethylthiazol-2-yl)-2，5-diphenyltetrazolium bromide(MTT).The protein expression levels of caspase-3(CASP3)，cleaved CASP3，and Bcl-2 related X(BAX) were detected by Western blotting.The concentrations of inflammatory factors including interleukin(IL)-1β，-6 and13 were detected by enzyme linked immunosorbent assay(ELISA).The direct regulation effect of LINC00052 on miR-145 was analyzed by dual luciferase reporter gene experiment.Rescue experiment was used to verify the role of LINC00052 by targeting miR-145 in the injury of TNF-α-induced C-20/A4 cells.Statistical analysis was compared by the t and analysis of variance.Results The expression level of LINC00052 in 0，10 and 30 ng/ mL of TNF-α-induced C-20/A4 cells showed an upward trend，while miR-145 showed a downward trend，and their expression was negatively correlated(P＜0.05).Interfering with LINC00052 reduced the proliferation of TNF-α-induced C-20/A4 cells，increased the protein expression levels of CASP3，cleared CASP3 and BAX，and elevated the concentrations of IL-1β，IL-6 and IL-13(P＜0.05).miR-145 is a direct target gene of LINC00052，and interfering with LINC00052 upregulated the expression level of miR-145 in C-20/A4 cells(P＜0.05).Interfering with miR-145 partially reversed the effect of LINC00052 suppression on the proliferation，apoptosis and inflammation in TNF-α-induced C-20/A4 cells(P＜0.05).Conclusion LINC00052 plays a protective role against the injury ofTNF-α-induced human articular chondrocytes by targeting miR-145，and its mechanism may be related to the promotion of proliferation and the inhibition of apoptosis and inflammation.

Key words： LINC00052 chondrocytes injury miR-145 regulation

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