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Role of cysteine‑rich angiogenic inducer 61 in fibroblast‑like synovial cell proliferation and invasion in rheumatoid arthritis.

Jie LG, Huang RY, Sun WF, Wei S, Chu YL, Huang QC, Du HY - Mol Med Rep (2014)

Bottom Line: The study found that the Cyr61 gene was highly expressed in the synovial cells from patients with RA compared with those from patients with OA.Conversely, overexpression of Cyr61 in normal FLS cells led to opposite effects.In conclusion, these results indicate that Cyr61 is capable of promoting RA‑FLS cell proliferation and invasion via the suppression of apoptosis and the regulation of MMP expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Chinese Medicine, Guangzhou General Hospital of Guangzhou Command of PLA, Guangzhou, Guangdong 510010, P.R. China.

ABSTRACT
Cysteine‑rich angiogenic inducer 61 (Cyr61) is a novel molecule that has been shown to be increased in the synovial tissues of patients with rheumatoid arthritis (RA). The present study was conducted in order to investigate the role of Cyr61 in the pathogenesis of RA. A human genome‑wide gene assay was used to screen gene expression in synovial tissues obtained from four patients with RA and three patients with osteoarthritis (OA). To examine the role of Cyr61 in the phenotype of RA‑fibroblast‑like synovial (FLS) cells, Cyr61 expression in RA‑FLS cells was knocked down using small interfering RNA (siRNA). Normal FLS cells transduced with lentiviral vectors encoding Cyr61 cDNA were used to further explore the effects of this molecule on FLS cell apoptosis, proliferation and invasion. The study found that the Cyr61 gene was highly expressed in the synovial cells from patients with RA compared with those from patients with OA. Downregulation of Cyr61 by siRNA led to impaired cell proliferation and invasion. Furthermore, it decreased the levels of matrix metalloproteinase (MMP)‑3 and MMP‑13, and induced apoptosis in RA‑FLS cells. Conversely, overexpression of Cyr61 in normal FLS cells led to opposite effects. In conclusion, these results indicate that Cyr61 is capable of promoting RA‑FLS cell proliferation and invasion via the suppression of apoptosis and the regulation of MMP expression. Therefore, Cyr61 may be a good target molecule for the treatment and prevention of RA.

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Role of Cyr61 in FLS cell apoptosis. (A) Fluorometric image of apoptotic cells. Blue, DAPI staining; Green, green fluorescent protein-positive staining for apoptosis. (B) Apoptosis ratio was calculated as indicated in the Materials and methods section. Data are expressed as the mean ± standard deviation of three independent experiments. *P<0.05 and **P<0.01. RA, rheumatoid arthritis; FLS, fibroblast-like synoviocytes; siRNA, small interfering RNA; NOR-FLS, normal FLS cells; NOR-CYR61, normal FLS cells transduced with lentivirus vector encoding Cyr61 cDNA; NOR-NC, normal FLS cells transduced with control lentivirus vector; RA-NC, RA-FLS cells transfected with control siRNA; RA-siRNA, RA-FLS cells transfected with Cyr61-siRNA; TUNEL assay, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling.
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f3-mmr-11-02-0917: Role of Cyr61 in FLS cell apoptosis. (A) Fluorometric image of apoptotic cells. Blue, DAPI staining; Green, green fluorescent protein-positive staining for apoptosis. (B) Apoptosis ratio was calculated as indicated in the Materials and methods section. Data are expressed as the mean ± standard deviation of three independent experiments. *P<0.05 and **P<0.01. RA, rheumatoid arthritis; FLS, fibroblast-like synoviocytes; siRNA, small interfering RNA; NOR-FLS, normal FLS cells; NOR-CYR61, normal FLS cells transduced with lentivirus vector encoding Cyr61 cDNA; NOR-NC, normal FLS cells transduced with control lentivirus vector; RA-NC, RA-FLS cells transfected with control siRNA; RA-siRNA, RA-FLS cells transfected with Cyr61-siRNA; TUNEL assay, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling.

Mentions: The data obtained by the MTS assay implied that Cyr61 may have a pro-survival effect via promotion of FLS cell proliferation. Subsequent experiments were conducted to examine whether the status of Cyr61 expression in FLS cells affects apoptosis. Analysis of the proportion of apoptotic cells in each group was performed using the fluorometric TUNEL method. As shown in Fig. 3A, knockdown of Cyr61 in RA-FLS cells by siRNA led to increased cell apoptosis compared with RA-FLS cells without transfection (2.8-fold) or RA-FLS cells transfected with control siRNA (2.7-fold; P<0.0001 and P<0.01, respectively; Fig. 3B). The proportion of apoptotic RA-FLS cells was significantly less than that of normal FLS cells (P<0.05; Fig. 3B). Transduction of normal FLS cells with lentiviral vectors encoding Cyr61 cDNA decreased the fraction of apoptotic cells by 86.5±0.01% compared with normal FLS cells, and by 89.8±0.02% compared with normal FLS cells transduced with control lentiviral vectors (P<0.05 and P<0.01, respectively; Fig. 3B).


Role of cysteine‑rich angiogenic inducer 61 in fibroblast‑like synovial cell proliferation and invasion in rheumatoid arthritis.

Jie LG, Huang RY, Sun WF, Wei S, Chu YL, Huang QC, Du HY - Mol Med Rep (2014)

Role of Cyr61 in FLS cell apoptosis. (A) Fluorometric image of apoptotic cells. Blue, DAPI staining; Green, green fluorescent protein-positive staining for apoptosis. (B) Apoptosis ratio was calculated as indicated in the Materials and methods section. Data are expressed as the mean ± standard deviation of three independent experiments. *P<0.05 and **P<0.01. RA, rheumatoid arthritis; FLS, fibroblast-like synoviocytes; siRNA, small interfering RNA; NOR-FLS, normal FLS cells; NOR-CYR61, normal FLS cells transduced with lentivirus vector encoding Cyr61 cDNA; NOR-NC, normal FLS cells transduced with control lentivirus vector; RA-NC, RA-FLS cells transfected with control siRNA; RA-siRNA, RA-FLS cells transfected with Cyr61-siRNA; TUNEL assay, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4262486&req=5

f3-mmr-11-02-0917: Role of Cyr61 in FLS cell apoptosis. (A) Fluorometric image of apoptotic cells. Blue, DAPI staining; Green, green fluorescent protein-positive staining for apoptosis. (B) Apoptosis ratio was calculated as indicated in the Materials and methods section. Data are expressed as the mean ± standard deviation of three independent experiments. *P<0.05 and **P<0.01. RA, rheumatoid arthritis; FLS, fibroblast-like synoviocytes; siRNA, small interfering RNA; NOR-FLS, normal FLS cells; NOR-CYR61, normal FLS cells transduced with lentivirus vector encoding Cyr61 cDNA; NOR-NC, normal FLS cells transduced with control lentivirus vector; RA-NC, RA-FLS cells transfected with control siRNA; RA-siRNA, RA-FLS cells transfected with Cyr61-siRNA; TUNEL assay, terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling.
Mentions: The data obtained by the MTS assay implied that Cyr61 may have a pro-survival effect via promotion of FLS cell proliferation. Subsequent experiments were conducted to examine whether the status of Cyr61 expression in FLS cells affects apoptosis. Analysis of the proportion of apoptotic cells in each group was performed using the fluorometric TUNEL method. As shown in Fig. 3A, knockdown of Cyr61 in RA-FLS cells by siRNA led to increased cell apoptosis compared with RA-FLS cells without transfection (2.8-fold) or RA-FLS cells transfected with control siRNA (2.7-fold; P<0.0001 and P<0.01, respectively; Fig. 3B). The proportion of apoptotic RA-FLS cells was significantly less than that of normal FLS cells (P<0.05; Fig. 3B). Transduction of normal FLS cells with lentiviral vectors encoding Cyr61 cDNA decreased the fraction of apoptotic cells by 86.5±0.01% compared with normal FLS cells, and by 89.8±0.02% compared with normal FLS cells transduced with control lentiviral vectors (P<0.05 and P<0.01, respectively; Fig. 3B).

Bottom Line: The study found that the Cyr61 gene was highly expressed in the synovial cells from patients with RA compared with those from patients with OA.Conversely, overexpression of Cyr61 in normal FLS cells led to opposite effects.In conclusion, these results indicate that Cyr61 is capable of promoting RA‑FLS cell proliferation and invasion via the suppression of apoptosis and the regulation of MMP expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Chinese Medicine, Guangzhou General Hospital of Guangzhou Command of PLA, Guangzhou, Guangdong 510010, P.R. China.

ABSTRACT
Cysteine‑rich angiogenic inducer 61 (Cyr61) is a novel molecule that has been shown to be increased in the synovial tissues of patients with rheumatoid arthritis (RA). The present study was conducted in order to investigate the role of Cyr61 in the pathogenesis of RA. A human genome‑wide gene assay was used to screen gene expression in synovial tissues obtained from four patients with RA and three patients with osteoarthritis (OA). To examine the role of Cyr61 in the phenotype of RA‑fibroblast‑like synovial (FLS) cells, Cyr61 expression in RA‑FLS cells was knocked down using small interfering RNA (siRNA). Normal FLS cells transduced with lentiviral vectors encoding Cyr61 cDNA were used to further explore the effects of this molecule on FLS cell apoptosis, proliferation and invasion. The study found that the Cyr61 gene was highly expressed in the synovial cells from patients with RA compared with those from patients with OA. Downregulation of Cyr61 by siRNA led to impaired cell proliferation and invasion. Furthermore, it decreased the levels of matrix metalloproteinase (MMP)‑3 and MMP‑13, and induced apoptosis in RA‑FLS cells. Conversely, overexpression of Cyr61 in normal FLS cells led to opposite effects. In conclusion, these results indicate that Cyr61 is capable of promoting RA‑FLS cell proliferation and invasion via the suppression of apoptosis and the regulation of MMP expression. Therefore, Cyr61 may be a good target molecule for the treatment and prevention of RA.

Show MeSH
Related in: MedlinePlus