Effects of miR-335-5p on proliferation and apoptosis of osteoarthritic chondrocytes by targeting programmed cell death 5

doi:10.3969/j.issn.2095-4344.3133

Abstract

Abstract: BACKGROUND: Studies have found that miR-335-5p is down-regulated in the synovial fluid of patients with osteoarthritis, which may be related to the occurrence and development of osteoarthritis.
OBJECTIVE: To investigate the effects of miR-335-5p on the proliferation and apoptosis of chondrocytes in osteoarthritis and the underlying mechanism.
METHODS: Human primary articular chondrocytes were isolated and cultured, and the cells were then treated with 10 μg/L interleukin-1β to establish an osteoarthritis model. The treated cells were divided into normal group (normal culture medium), interleukin-1β group, miR-NC group, miR-335-5p group, si-NC group, si-programmed cell death 5 (PDCD5) group, miR-335-5p+pcDNA3.1 group, and miR-335-5p+pcDNA3.1-PDCD5 group. After 24, 48, and 72 hours of culture, the cell proliferation and apoptosis rate were detected by MTT and flow cytometry, respectively. The mRNA expression levels of miR-335-5p, PDCD5, ADAMTS-5 and matrix metalloproteinase-13 in chondrocytes were detected by qRT-PCR. The expression levels of PDCD5, Cyclin D1, p21, Bax and Bcl-2 proteins in chondrocytes were determined by western blot. The relationship between miR-335-5p and PDCD5 was verified by double luciferase reporting assay system. The study procedures were implemented in line with the relevant ethic requirements of the First Hospital of Changsha.
RESULTS AND CONCLUSION: Compared with the control group, interleukin-1β inhibited the cell proliferation and promoted the apoptosis of chondrocytes, and inhibited the expression of miR-335-5p in chondrocytes. Over-expression of miR-335-5p promoted the proliferation and inhibited the apoptosis of chondrocytes induced by interleukin-1β. miR-335-5p targeted and negatively regulated the expression of PDCD5. Inhibition of PDCD5 promoted the proliferation of chondrocytes induced by interleukin-1β. Overexpression of PDCD5 reversed the effects of miR-335-5p up-regulation on the proliferation and apoptosis of chondrocytes induced by interleukin-1β. To conclude, miR-335-5p promotes cell proliferation and inhibits cell apoptosis by targeting PDCD5. miR-335-5p is a potential molecular target for osteoarthritis.

Key words: osteoarthritis, chondrocyte, miR-335-5p, PDCD5, proliferation, apoptosis

CLC Number:

Zhang Yunqing, Wang Jian, Yang Chunhua, Li Cong. Effects of miR-335-5p on proliferation and apoptosis of osteoarthritic chondrocytes by targeting programmed cell death 5[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(14): 2142-2147.

Figures/Tables 7

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