Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 1. Immunostaining of PinX1 in CRC tissues. (A) Intensity of staining in CRC tissues. Top panel, x100 magnification; bottom panel, x200 magnification; a and e, negative; b and f, weak; c and g, moderate; d and h, strong. (B) The distribution of different staining intensities of PinX1 in CRC tissues compared with adjacent non‑cancerous control tissues (P<0.001, paired Wilcoxon signed‑rank test). (C) Low PinX1 expression is associated with lymph node metastasis (*P=0.021, χ2 test). (D) Low PinX1 expression is associated with distant metastasis (*P=0.030, χ2 test). (E) Low PinX1 expression is associated with advanced Tumor‑Node‑Metastasis stage (*P=0.014, χ2 test). PinX1, Pin2/TRF1‑binding protein X1; CRC, colorectal cancer; N‑C, adjacent non‑cancerous tissues‑CRC tissues; N, node; M, metastasis.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Immunostaining, Staining, Control, Expressing

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 2. Expression of PinX1 is associated with overall and disease‑free survival in patients with CRC. (A) Low PinX1 expression is associated with a poorer overall cumulative survival in patients with CRC (P=0.001, log‑rank test). (B) Low PinX1 expression is associated with a poorer disease‑free cumulative survival in patients with CRC (P=0.017, log‑rank test). Cum, cumulative; PinX1, Pin2/TRF1‑binding protein X1.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Expressing

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 3. PinX1 inhibits the migration and invasion of colorectal cancer cells. (A) The relative protein expression level of PinX1 in PinX1‑knockdown (shPinX1) and control groups (shCtrl) in HCT116 and SW480 cells was detected by western blotting. (B) Densitometric analysis of the relative protein expression level of PinX1 in PinX1‑knockdown (shPinX1) and control groups (shCtrl) in HCT116 and SW480 cells. (C and D) PinX1‑knockdown inhibited the migration ability of HCT116 and SW480 cells. (E and F) PinX1‑knockdown inhibited the invasion ability of HCT116 and SW480 cells. (G and H) PinX1‑knockdown inhibited the proliferation ability of HCT116 and SW480 cells. All experiments were performed in triplicate. Data are presented as the mean ± standard deviation. ***P<0.001; **P<0.01. PinX1, Pin2/TRF1‑binding protein X1; sh, short hairpin RNA; Ctrl, control.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Migration, Expressing, Control, Western Blot, Standard Deviation, shRNA

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 4. PinX1 expression inhibits the expression and activity of MMP2, and inhibits expression of p65 and p‑p65. (A) Western blot analysis of the relative protein levels of PinX1, MMP2, MMP9, TIMP‑1 and TIMP‑2 in HCT116 and SW480. (B) Gelatin zymography analysis of MMP2 in HCT116 and SW480. (C) Western blot analysis of the relative protein levels of PinX1, MMP2, p65 and p‑p65 in HCT116 and SW480. All experiments were performed in triplicate through comparing knockdown of the PinX1 group (shPinX1) with that of the control group (shCtrl). Histograms represent the mean ± standard deviation. ***P<0.001. PinX1, Pin2/TRF1‑binding protein X1; MMP2, matrix metalloproteinase 2; p‑, phosphorylated; TIMP, TMP metallopeptidase inhibitor 1; sh, short hairpin RNA; Ctrl, control.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Expressing, Activity Assay, Western Blot, Zymography, Knockdown, Control, Standard Deviation, shRNA

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 5. PinX1 suppresses MMP2 expression via the nuclear factor‑κB pathway. Western blot analysis of the relative protein expression levels of PinX1, MMP2, p65 and p‑p65 in shCtrl and shPinX1 groups and those co‑treated with p65 siRNA in (A) HCT116 and (B) SW480 cells. The p65‑specific siRNA efficiently prevented the upregulation of MMP2 expression induced by the knockdown of PinX1. (C and D) The improved migration and invasion ability resulting from the knockdown of PinX1 was inhibited by p65‑siRNA in HCT116 and SW480 cells. All experiments were performed in triplicate. Histograms represent the mean ± standard deviation. **P<0.01; ***P<0.001. PinX1, Pin2/TRF1‑binding protein X1; MMP2, matrix metalloproteinase 2; p‑, phosphorylated; si, small interfering; sh, short hairpin RNA; Ctrl, control.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Expressing, Western Blot, Knockdown, Migration, Standard Deviation, shRNA, Control

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 6. PinX1 negatively regulates colorectal cancer cell metastasis in vivo. (A) Right panel, representative image of liver with metastatic nodules; middle panel, H&E staining sections of liver with metastatic nodules; left panel, representative image of H&E stained lung sections. (B) Representative image of 10% formalin‑fixed liver with metastatic nodules. (C) The number of liver metastatic nodules was counted under a dissecting microscope. Compared with the shCtrl group, a statistically significant increase in the number of the liver metastases was observed in the shPinX1 group (***P=0.002). (D) Immunohistochemical staining of MMP2 and p65 in metastatic liver nodules. MMP2 and p65 expression was increased in the shPinX1 group, compared with the shCtrl group. sh, short hairpin; Ctrl, control; PinX1, Pin2/TRF1‑binding protein X1; MMP2, matrix metalloproteinase 2.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: In Vivo, Staining, Microscopy, Immunohistochemical staining, Expressing, Control