Fig 1: Cyr61 expression is elevated in OSCC and associated with advanced clinicopathological features. (A) Representative immunohistochemical staining of Cyr61 in OSCC tissue specimens from stages I-IV, together with the negative control. (B) Quantification of Cyr61-positive staining area in OSCC specimens according to clinical stage. (C) Cyr61 mRNA expression in normal oral mucosa and OSCC tissues from the GEO dataset GSE30784. (D) Kaplan-Meier overall survival analysis of the TCGA oral cavity/OSCC subset stratified by median Cyr61 expression, with annotation of the log-rank p value and the corresponding univariate and multivariate Cox regression results. (E) Forest plot of multivariate Cox proportional hazards analysis in the TCGA oral cavity/OSCC subset, showing hazard ratios for Cyr61 expression, clinical stage, age, and sex. (F) Cyr61 mRNA expression in metastatic lymph node samples and primary tumor samples from OSCC patients in the GEO dataset GSE2280. *p < 0.05 as indicated.
Fig 2: PLC/PKC/c-Src signaling is required for Cyr61-induced ICAM-1 expression and OSCC migration. (A) Phosphorylation of PLC, PKC, and c-Src in SCC4 and SAS cells following Cyr61 stimulation (n = 4). (B-D) Quantification of (B) cell movement, (C) cell migration, and (D) ICAM-1 mRNA expression in cells pretreated with inhibitors of PLC, PKC, or c-Src (U73122, 1 µM; GF109203X, 5 µM; or PP2, 5 µM), followed by Cyr61 stimulation (30 ng/mL) (n = 4). (E-G) Quantification of (E) cell movement, (F) cell migration, and (G) ICAM-1 mRNA expression following siRNA-mediated knockdown of PLC, PKC, or c-Src and Cyr61 stimulation (30 ng/mL) (n = 4). Data are presented as mean ± SD of four independent experiments. *p < 0.05 compared with the control group; #p < 0.05 compared with the Cyr61-treated group.
Fig 3: AP-1/c-Jun signaling mediates Cyr61-induced ICAM-1 expression and OSCC migration. (A) Phosphorylation of c-Jun following Cyr61 stimulation (n = 4). (B-D) Quantification of (B) cell movement, (C) cell migration, and (D) ICAM-1 mRNA expression in cells pretreated with curcumin (1 μM) or tanshinone IIA (5 μM) for 1 h before stimulation with Cyr61 (30 ng/mL) (n = 4). (E-G) Quantification of (E) cell movement, (F) cell migration, and (G) ICAM-1 mRNA expression following c-Jun siRNA transfection and Cyr61 stimulation (30 ng/mL) (n = 4). Data are presented as mean ± SD of four independent experiments. *p < 0.05 compared with the control group; #p < 0.05 compared with the Cyr61-treated group.
Fig 4: Cyr61 activates AP-1 and enhances c-Jun binding to the ICAM-1 promoter in OSCC cells. (A) AP-1 luciferase activity following Cyr61 stimulation (3-30 ng/mL) (n = 4). (B) AP-1 luciferase activity in cells pretreated with an anti-integrin αvβ3 antibody or with inhibitors of PLC, PKC, or c-Src (U73122, 1 µM; GF109203X, 5 µM; or PP2, 5 µM), followed by Cyr61 stimulation (30 ng/mL) (n = 4). (C) c-Jun phosphorylation in cells pretreated with PLC, PKC, or c-Src inhibitors prior to Cyr61 stimulation (30 ng/mL) (n = 4). (D) Chromatin immunoprecipitation (ChIP) analysis showing c-Jun binding to the AP-1 recognition sequence within the ICAM-1 promoter following pretreatment with specific inhibitors and Cyr61 stimulation (30 ng/mL) (n = 4). Data are presented as mean ± SD of four independent experiments. *p < 0.05 compared with the control group; #p < 0.05 compared with the Cyr61-treated group.
Fig 5: Cyr61 enhances OSCC motility and migration through upregulation of ICAM-1 expression. (A, B) OSCC cells were treated with recombinant Cyr61 at the indicated concentrations for 24 h, and (A) cell movement (wound-healing assay) and (B) cell migration (Transwell assay) were assessed (n = 4). (C) Quantification of cell migration in OSCC cells preincubated with control IgG or anti-Cyr61 antibody (100 pg/mL) for 24 h (n = 4). (D) Correlation analysis between Cyr61 and ICAM-1 expression in the TCGA-HNSC cohort using TIMER2.0. Spearman correlation. (E) ICAM-1 mRNA expression following Cyr61 treatment (10-30 ng/mL) (n = 4). (F) ICAM-1 protein expression following Cyr61 treatment at the indicated concentrations (n = 4). (G, H) Quantification of (G) cell movement and (H) cell migration following control and ICAM-1 siRNA transfection and Cyr61 stimulation (30 ng/mL) (n = 4). Data are presented as mean ± SD of four independent experiments. *p < 0.05 compared with the control group; #p < 0.05 compared with the Cyr61-treated group.
Supplier Page from Abcam for Human CYR61 ELISA Kit