JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2015, Vol. 53 ›› Issue (12): 20-26.doi: 10.6040/j.issn.1671-7554.0.2014.917

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Effects of PCSK6 silence by siRNA on FLS in CIA

JIANG Huiyu1,2,3,4, WANG Lin1,2,3, PAN Jihong1,2,3   

  1. 1. Shandong Medicinal Biotechnology Center, Jinan 250062, Shandong, China;
    2. Key Laboratory for Biotech-Drugs Ministry of Health, Jinan 250062, Shandong, China;
    3. Key Laboratory for Rare & Uncommon Diseases of Shandong Province, Jinan 250062, Shandong, China;
    4. School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, Jinan 250200, Shandong, China
  • Received:2014-12-08 Online:2015-12-10 Published:2015-12-10

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Abstract: Objective To explore the effects of PCSK6 silence by siRNA on the biological behaviors of fibroblast-like synoviocytes (FLS) in collagen-induced arthritis (CIA). Methods Rat CIA models were induced with bovine collagen type Ⅱ. Cultured FLS from CIA models were treated with anti-PCSK6 siRNA, including siRNA oligo-433, siRNA oligo-688 and siRNA oligo-2506. A negative siRNA was used as the negative control. The inhibiting effects were detected with RT-qPCR after 24 h and 36 h. The proliferation, invasion, migration capacity, secretion of inflammation factors and cell cycle were determined with MTT, transwell, wound healing, ELISA and flow cytomertry. Expressions of genes related to proliferation, invasion, migration and inflammation were detected with RT-qPCR. Results Compared with negative control, the inhibiting effect was remarkable after anti-PCSK6 siRNA (siRNA oligo-2506) was transfected into FLS at 24 h (P<0.001). PCSK6-siRNA treatment significantly decreased cell proliferation (P<0.001), invasion (P<0.001) and migration (P<0.001) of FLS in CIA, as well as secretion of tumor necrosis factor alpha (TNF-α) and IL-1β (P<0.001). Flow cytometry revealed a G0/G1 arrest of FLS (P<0.05). Expression of genes, including CXCL9, MMP-2, MMP-9, NOSTRIN, HIF-1α, MPZL2, IGF-2 and PADI4 all decreased significantly(P<0.05). Conclusion Inhibition of PCSK6 may play a protective role in the biological behaviors of FLS in CIA, which may provide information for further study of the pathogenesis of CIA.

Key words: Collagen-induced arthritis, Rheumatoid arthritis, PCSK6, siRNA, Fibroblast-like synoviocytes

CLC Number: 

  • R593.22
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