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

Previous Articles     Next Articles

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

PDF (PC)

380

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
[1] Sylvie S, Peggy J, Jens VP, et al. A multiparameter approach to monitor disease activity in collagen-induced arthritis[J]. Arthritis Res Ther, 2010, 12(4):160.
[2] Ben Hassine H, Zemni R, Bouagina E, et al. Lack of association between PADI4 polymorphisms and rheumatoid arthritis in the Tunisian population[J]. Joint Bone Spine, 2012, 79(3):329-330.
[3] 张玲玲,沈玉先,魏伟.类风湿关节炎的关节滑膜病理生理机制[J].生理科学进展, 2003, 34(2):144-146.
[4] Trentham DE, Townes AS, Kang AH. Autoimmunity to typeⅡcollagen:an experimental model of arthritis[J]. J Exp Med, 1977, 146(3):857-868.
[5] Holmdahl R, Mo J, Nordling C, et al. Collagen induced arthritis:an experimental model for rheumatoid arthritis with involvement of both DTH and immune complex mediated mechanisms[J]. Clin Exp Rheumatol, 1989, 7(Suppl 3):51-55.
[6] 张敏娜.类风湿关节炎的实验动物模型及其评价[J].实验动物科学, 2011, 28(4):57-59.
[7] Lin H, Ah Kioon MD, Lalou C, et al. Protective role of systemic furin in immune response-induced arthritis[J]. Arthritis Rheum, 2012, 64(9):2878-2886.
[8] Wang F, Wang L, Jiang H, et al. Inhibition of PCSK6 may play a protective role in the development of rheumatoid arthritis[J]. J Rheumatol, 2015, 42(2):161-169.
[9] Seidah NG, Khatib AM, Prat A. The proprotein convertases and their implication in sterol and/or lipid metabolism[J]. Biol Chem, 2006, 387(7):871-877.
[10] 徐华,马韵,龙喜带.前蛋白转化酶与恶性肿瘤[J].肿瘤防治研究, 2013, 40(1):105-110.
[11] Mains RE, Berard CA, Denault JB, et al. PACE4:a subtilisin-like endoprotease with unique properties[J]. Biochem J, 1997, 321(Pt 3):587-593.
[12] Nour N, Mayer G, Mort JS, et al. The cysteine-rich domain of the secreted proprotein convertases PC5A and PACE4 function as a cell surface anchor and interacts with tissue inhibitors of metalloproteinases[J]. Mol Biol Cell, 2005, 16(11):5215-5226.
[13] 李建平,王晓滨,陈常忠,等.偶合基本氨基酸裂解酶4基因多态性与高血压的相关性研究[J].中华心血管病杂志, 2004, 32(9):781-784. LI Jianping, WANG Xiaobin, CHEN Changzhong, et al. The association of a polymorphism in the paired basic amino acid cleaving enzyme 4 gene and hypertension[J]. Chin J Cardiol, 2004, 32(9):781-784.
[14] Malfait AM, Seymour AB, Gao F, et al. A role for PACE4 in osteoarthritis pain:evidence from human genetic association and null mutant phenotype[J]. Ann Rheum Dis, 2012, 71(6):1042-1048.
[15] 刘元刚,刘树滔,饶平凡.类风湿关节炎啮齿动物模型的研究进展[J].中国实验动物学报, 2007, 15(6):470-473. LIU Yuangang, LIU Shutao, RAO Pingfan. Recent advances in research on rodent models of rheumatoid arthritis[J]. Acta Lab Anim Sci Sin, 2007, 15(6):470-473.
[16] 许金鑫,张育,张学增,等.金雀异黄素对CIA大鼠炎症及细胞因子影响的研究[J].实用临床医药杂志, 2010, 14(19):1-5. XU Jinxin, ZHANG Yu, ZHANG Xuezeng, et al. Effect of Gen on the inflammation and cytokines of rats with collagen type Ⅱ induced arthritis[J]. Journal of Clinical Medicine in Practice, 2010, 14(19):1-5.
[17] Lee A, Qiao Y, Grigoriev G, et al. TNFα induces sustained signaling and a prolonged and unremitting inflammatory response in synovial fibroblasts[J]. Arthritis Rheum, 2013, 65(4):928-938.
[18] Feldmann M, Maini RN. Lasker Clinical Medical Research Award. TNF defined as a therapeutic target for rheumatoid arthritis and other autoimmune diseases[J]. Nat Med, 2003, 9(10):1245-1250.
[19] Joosten LA, Helsen MM, van de Loo FA, et al. Anticytokine treatment of established type Ⅱ collagen-induced arthritis in DBA/1 mice:a comparative study using anti-TNFalpha, anti-IL-1alpha/beta and IL-1Ra[J]. Arthritis Rheum, 2008, 58(2 suppl):110-122.
[20] Kuan WP, Tam LS, Wong CK, et al. CXCL9 and CXCL10 as sensitive markers of disease activity in patients with rheumatoid arthritis[J]. J Rheumatol, 2010, 37(2):257-264.
[21] Pardo A, Selman M. Matrix metalloproteases in aberrant fibrotic tissue remodeling[J]. Proc Am Thorac Soc, 2006, 3(4):383-388.
[22] Kovacevic I, Hu J, Siehoff-Icking A, et al. The F-BAR protein NOSTRIN participates in FGF signal transduction and vascular development[J]. EMBO J, 2012, 31(15):3309-3322.
[23] Hollander AP, Corke KP, Freemont AJ, et al. Expression of hypoxia-inducible factor 1alpha by macrophages in the rheumatoid synovium:implications for targeting of therapeutic genes to the inflamed joint[J]. Arthritis Rheum, 2001, 44(7):1540-1544.
[1] LI Jian, XU Bing, YAN Xinfeng, XU Wanju, CHANG Xiaotian. Screening key genes for TXNDC5 and insulin signaling pathway [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(3): 88-93.
[2] XIA Chuanyou, LI Luchao, LI Xiaofeng, YAN Keqiang, ZHANG Nianzhao, FAN Yidong, LIU Cheng. Expression of histone methyltransferase SMYD3 in prostate cancer tissues and its role in mTOR pathway [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(5): 12-16.
[3] ZHAO Yan, ZHENG Yabing, YAN Xinfeng, ZHANG Hu, CHANG Xiaotian. Screening crucial genes for glucose metabolism in rheumatoid arthritis [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(3): 30-35.
[4] ZHANG Rongjun, HAN Bo, GAO Ling, ZHU Mei, DING Guoyu, LIANG Yan. Effects of CD40 knockdown by siRNA on rats with experimental autoimmune myocarditis and IL-22 expression [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2015, 53(5): 36-40.
[5] ZHENG Jiatian, ZHU Kai, SUN Hongsheng, FU Min, PAN Zhenglun. Association of IL-18 and MCP-1 gene polymorphisms with rheumatoid arthritis: a meta-analysis [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(6): 98-104.
[6] DING Ling1,2, GONG Huiping1, DU Yimeng1, WANG Xin1, ZHANG Yan3, WANG Xuping2, LU Qinghua1. Effect of SIRT3 siRNA on ICAM-1 and E-selectin expressions in HAEC stimulated by ox-LDL [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(4): 43-48.
[7] LI Yan, WEI Wei, LV Shuqing, TIAN Yongjie. Effects of PARP-1 on angiogenesis of ovarian cancer [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(4): 97-101.
[8] WANG Jie-zhong1, HAN Bo1, GAO Ling2, YANG Wen-wei1, LIU Feng-qin1, Lv-Jian-li1, Zhao Li-jian1. Effect of CD40siRNA on CD4+CD25+ regulatory T cells in rats with experimental autoimmune myocarditis [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(1): 1-4.
[9] LI Feng1, XU Jing2, ZHENG Jia-tian1, ZHANG Yuan-chao1， SUN Hong-sheng1, PAN Zheng-lun1. Association between CD14 gene polymorphisms and rheumatoid arthritis in Han population of Shandong province [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(4): 87-91.
[10] LIU Hong-li, ZHANG You-zhong, Lv Chang-shuai, DING Bai-juan, WANG Jie. Effect of nucleolin down-regulation on biological behaviors of HeLa cells in human cervical carcinoma [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(11): 46-50.
[11] LIANG Ting-ting， ZHANG Ping， MA De-mei， ZHU Lin. Effect on apoptosis mediated by TRAIL against Caski cells of HPV16-E5-targeted siRNA [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(10): 24-28.
[12] ZHANG Yu-jun, DING Feng，WANG Chun-xiao， LI Xing-fu，YU Xiao. Effects of 99Tc-MDP on PGE1 and PGE2 in rats of adjuvant arthritis [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2010, 48(8): 50-53.
[13] WEI Chao1, WU Wei-ping1, ZHAO Yan1, WANG Lin1, YAN Xin-feng2, CHANG Xiao-tian1 . Effects of estrogen on the disease development in the collagen-induced arthritis  animal models [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2010, 48(8): 54-59.
[14] ZHUANG Xue-wei,XIA xi-yan,SHAN Ning-ning,WANG Hong-chun,ZHANG Yi,YANG Xiao-jing,LI Xiao-li,ZHAO Sheng-mei,ZOU Xiong. Construction of a plasmid expressing siRNA aimed at the H-2Kd gene and expression of H-2Kd in mouse LAK cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2008, 46(6): 590-593.
[15] SHAN Ning-ning,ZOU Xiong,ZHANG Yi,YANG Xiao-jing,ZHUANG Xue-wei,WANG Hong-chun,HOU Ming. Silence of MHC gene has an effect on the cytotoxicity of LAK cells by small interfering RNA duplexes in mice [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2007, 45(11): 1096-1100.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd