Volume 19 Issue 5
May  2021
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Article Navigation >  Chinese Journal of General Practice  > 2021  >  19(5): 745-748,874
WU Jie, FAN Wen-qiang, XU Zhen-dan, LIANG Shu, QIN Yi-lu, WANG Pei-shan. Study on the molecular mechanism of miR-335-5p on proliferation and apoptosis of osteoarthritis chondrocytes[J]. Chinese Journal of General Practice, 2021, 19(5): 745-748,874. doi: 10.16766/j.cnki.issn.1674-4152.001906
Citation: WU Jie, FAN Wen-qiang, XU Zhen-dan, LIANG Shu, QIN Yi-lu, WANG Pei-shan. Study on the molecular mechanism of miR-335-5p on proliferation and apoptosis of osteoarthritis chondrocytes[J]. Chinese Journal of General Practice, 2021, 19(5): 745-748,874. doi: 10.16766/j.cnki.issn.1674-4152.001906
shu

Study on the molecular mechanism of miR-335-5p on proliferation and apoptosis of osteoarthritis chondrocytes

doi: 10.16766/j.cnki.issn.1674-4152.001906
  • 1.

    Department of Rheumatology and Immunology, Xinxiang Central Hospital, Xinxiang, Henan 453000, China

Funds:

 LHGJ20200941

 GG2019026

  • Received Date: 2020-08-04
    Available Online: 2022-02-16
    Abstract
  •   Objective  To explore the effect of miR-335-5p on the proliferation and apoptosis of osteoarthritis (OA) chondrocytes.  Methods  OA chondrocytes and normal chondrocytes were isolated and cultured in vitro. The expression of miR-335-5p in the cells was detected by qRT-PCR, and the expression of Tnfrsf1a protein was detected by western blotting. OA chondrocytes were divided into anti-miR-NC group, anti-miR-335-5p group, anti-miR-335-5p+pcDNA group, and anti-miR-335-5p+pcDNA-Tnfrsf1a group. MTT, flow cytometry and western blotting were used to detect proliferation, apoptosis and the protein expression of p53 and Survivin, respectively. Dual luciferase reporter assay verified the targeted regulation relationship between miR-335-5p and Tnfrsf1a. The comparison between the two groups was performed by independent sample t test, the comparison between multiple groups was performed by one-way analysis of variance, and the further comparison between the two groups was performed by LSD-t test.  Results  Compared with normal chondrocyte, the expression of miR-335-5p in OA chondrocytes reduced (0.31±0.08 vs. 1.00±0.00, t=25.875, P < 0.05), while the expression of Tnfrsf1a miRNA (6.54±0.30 vs. 1.00±0.00, t=55.400, P < 0.05) and protein (0.95±0.18 vs. 0.18±0.03, t=12.659, P < 0.05) increased. Compared with the anti-miR-NC group, the OD value of OA chondrocytes in the anti-miR-335-5p group decreased (0.27±0.01 vs. 0.34±0.01, t=14.849, P < 0.05), while the apoptosis rate increased [(19.18±0.88) % vs. (6.89±0.33) %, t=39.230, P < 0.05], and the expression of p53 protein increased (P < 0.05), but the expression of Survivin protein decreased (P < 0.05). miR-335-5p targeted and negatively regulated the expression of Tnfrsf1a. Compared with the miR-335-5p + si-NC group, the OD value of OA chondrocytes in the miR-335-5p + si-Tnfrsf1a group increased, while the apoptosis rate was decreased, and the expression of p53 protein decreased (P < 0.05), but the expression of Survivin protein increased (P < 0.05).  Conclusion  Knockdown of miR-335-5p could inhibit the proliferation and promote apoptosis of OA chondrocyte cells by negatively regulating Tnfrsf1a, and it may become a new target for OA treatment.

     

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