title: Inhibition of smoothened decreases proliferation of synoviocytes in rheumatoid arthritis.
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 G1phase 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.
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.|Center for Clinic Immunology, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Shang-Ling
authors.initials: SL
authors.lastname: Zhu
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.|Center for Clinic Immunology, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Jian-Lin
authors.initials: JL
authors.lastname: Huang
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Wei-Xiang
authors.initials: WX
authors.lastname: Peng
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Dan-Chun
authors.initials: DC
authors.lastname: Wu
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Min-Qi
authors.initials: MQ
authors.lastname: Luo
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Qiu-Xia
authors.initials: QX
authors.lastname: Li
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Zhao-Xia
authors.initials: ZX
authors.lastname: Li
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Xiao-Xue
authors.initials: XX
authors.lastname: Feng
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.affiliation: Division of Rheumatology, Department of Internal Medicine, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Ming-Xia
authors.initials: MX
authors.lastname: Wang
authors.affiliation: Division of Rheumatology, Penn State University Hershey College of Medicine, Hershey, PA, USA.
authors.forename: Wei-Qian
authors.initials: WQ
authors.lastname: Chen
authors.affiliation: Division of Rheumatology, Penn State University Hershey College of Medicine, Hershey, PA, USA.
authors.forename: Nancy
authors.initials: N
authors.lastname: Olsen
authors.affiliation: Center for Clinic Immunology, the Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong Province, People's Republic of China.|Division of Rheumatology, Penn State University Hershey College of Medicine, Hershey, PA, USA.
authors.forename: Song Guo
authors.initials: SG
authors.lastname: Zheng
mesh_terms: D017209:Apoptosis / Q000235:genetics; D001172:Arthritis, Rheumatoid / Q000473:pathology*; D049109:Cell Proliferation / Q000235:genetics; D005260:Female; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D059585:G1 Phase Cell Cycle Checkpoints / Q000235:genetics; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D000072086:Smoothened Receptor / Q000819:agonists / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000070918:Synoviocytes / Q000473:pathology*
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; C491023:SMO protein, human; D000072086:Smoothened Receptor