IL-17RB expression has an essential role in tumorigenesis and metastasis of pancreatic tumor cells. (A) Immunoblot analysis of IL-17RB in pancreatic cancer cell lines. α-Tubulin was used as a loading control. (B) IB analysis of IL-17RB in CFPAC-1 and BxPC3 cell lines transduced with lentiviral-control shRNA (shLacZ) or shIL-17RB. Tubulin was used as a loading control. RE, relative expression. (C and D) Soft agar colony formation (SACF) assay (C) and invasion assay (D) using CFPAC-1 and BxPC3 cells infected with shLacZ or shIL-17RB. (E) IB analysis of IL-17RB in control (neo), IL-17RB full-length or ΔLBD overexpressing SU86.86 and HPAC cell lines. Tubulin was used as a loading control. RE, relative expression. (F and G) Assays for soft agar colony formation (SACF; F) and invasion (G) were performed using SU.86.86 and HPAC cells overexpressing full-length or ΔLBD IL-17RB. (H) Tumorigenesis assay of NOD/SCIDγ mice subcutaneously injected with shLacZ-transduced or IL-17RB–depleted CFPAC-1 cells. Cell dose: 1 × 106 cells per mouse. Six mice were used for each group. (I) Tumor weight of NOD/SCIDγ mice orthotopically implanted with shLacZ-transduced or IL-17RB–depleted CFPAC-1 cells. Cell dose: 2.5 × 105 cells per mouse. Six mice were used for each group. (J) IHC of IL-17RB in tumors derived from mice orthotopically implanted with shLacZ-transduced or IL-17RB–depleted CFPAC-1 cells. (K) Summary table of lung and liver metastasis derived from orthotopic xenograft. (L) Lung metastasis of NOD/SCIDγ mice intravenously injected with shLacZ-transduced or IL-17RB–depleted CFPAC-1 cells. Cell dose: 5 × 105 cells per mouse. Six mice were used for each group. Data shown are means ± SD. *, P < 0.05; **, P < 0.01 (Student’s t test). All experimental data verified in at least two independent experiments.
