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Inhibition of smoothened decreases proliferation of synoviocytes in rheumatoid arthritis

Cellular & Molecular Immunology volume 14, pages 214–222 (2017)Cite this article

Abstract

Fibroblast-like synoviocytes (FLSs) contribute to synovial hyperplasia in rheumatoid arthritis (RA). Smoothened (Smo) is a key component of sonic hedgehog (Shh) signaling and contributes to tumor cell proliferation. The objective of this study was to investigate the role of Smo in RA synoviocyte proliferation. FLSs were isolated from RA synovium. Shh signaling was studied using a Smo antagonist (GDC-0449) and small interfering RNA (siRNA) targeting the Smo gene in FLSs. Cell proliferation was quantified by using kit-8 assay and cell cycle distribution and apoptosis were evaluated by flow cytometry. Cell cycle-related genes and proteins were detected by real-time PCR and western blot. FLSs treated with GDC-0449 or Smo-siRNA showed significantly decreased proliferation compared to controls (P < 0.05). Incubation with GDC-0449 or transfection with Smo-siRNA resulted in a significant increase of G1 phase cells compared to controls (P < 0.05). Cell cycle arrest was validated by the significant increase in cyclin D1 and E1 mRNA expression, decrease in cyclin-dependent kinase p21 mRNA expression in Smo-siRNA transfected cells (P < 0.05). Protein expression of cyclin D1 was also downregulated after Smo gene knockdown (P < 0.05). The results suggest that Shh signaling plays an important role in RA-FLSs proliferation in a Smo-dependent manner and may contribute to synovial hyperplasia. Targeting Shh signaling may help control joint damage in patients with RA.

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Acknowledgements

This work was supported in part by grants from the National Natural Science Foundation of China (81072480), from the Natural Science Foundation of Guangdong Province (S2012020010927), and from the Science and Technology Program of Guangdong Province (2013B021800076) (Jian-lin Huang); the major projects from Science and Technology Program of Guangzhou City, from the National Natural Science Foundation of Guangdong Province, from the Department of Education of Guangdong Province, and grants from NIH AR059103 and NIH AI084359 (Song Guo Zheng).

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Author notes

  1. Shang-ling Zhu and Jian-lin Huang: Both Shang-ling Zhu and Jian-lin Huang equally contributed to this work.

  2. Dr. JL Huang, Center for Clinic Immunology, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China. E-mail: jianlin_h@163.com

  3. Dr. SG Zheng, szheng1@hmc.psu.edu

Authors and Affiliations

  1. Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China

    Shang-ling Zhu, Jian-lin Huang, Wei-xiang Peng, Dan-chun Wu, Min-qi Luo, Qiu-xia Li, Zhao-xia Li, Xiao-xue Feng, Fang Liu & Ming-xia Wang

  2. Center for Clinic Immunology, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China

    Shang-ling Zhu, Jian-lin Huang & Song Guo Zheng

  3. Division of Rheumatology, Penn State University Hershey College of Medicine, Hershey, PA, USA

    Wei-qian Chen, Nancy Olsen & Song Guo Zheng

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Zhu, Sl., Huang, Jl., Peng, Wx. et al. Inhibition of smoothened decreases proliferation of synoviocytes in rheumatoid arthritis. Cell Mol Immunol 14, 214–222 (2017). https://doi.org/10.1038/cmi.2015.67

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