Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 1. PinX1 expression was observed in 26 pairs of breast cancer tissues and the normal counterparts. (A and B) Protein levels of PinX1, as assessed by immunoblot analysis with an anti-PinX1 antibody are downregulated in a majority of cancer tissues compared with the paired normal breast tissues (23/26). Separately, (B) showed significant differences (**P<0.01, two-tailed paired t-test) and (A) presented 12 pairs of the 26 pairs. GAPDH was a loading control; T, tumor; N, non-tumor. (C and D) Quantitative PCR analysis of PinX1 mRNA expression in 26 paired samples. Data are shown relative to those of the internal control gene Gapdh. (D) Significant differences (**P<0.01, two-tailed paired t-test) of PinX1 expression, (C) representing 15 pairs of the total.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Expressing, Western Blot, Two Tailed Test, Control, Real-time Polymerase Chain Reaction

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 2. Immunohistochemical staining of PinX1 was analysed in breast cancer tissues and the normal breast tissues. (A) A cancer adjacent normal breast tissue (case 1) with high expression of PinX1, where almost all tumor cells were positively stained (magnification, x100). (B) A fibroadenoma tissue (case 3) with >65% of tumor cells positive staining of PinX1 (magnification, x100). (C) An invasive ductal carcinoma (case 43) exhibited low expression of PinX1, where <45% of tumor cells were positive staining of PinX1 (magnification, x100). (G) Low expression of PinX1 was obtained in an invasive ductal carcinoma (case 29), where <10% of tumor cells were positive staining of PinX1 (magnification, x100). Respectively, (D-H) revealed the higher magnification (magnifica- tion, x400) of the specific areas boxed in (A, B, C and G).

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 3. The ROC curve was used to obtain the optimal cut-off value for positive and negative PinX1 expression. The plots of sensitivity and specificity for each clinicopathological parameter was as follows: (A) age at surgery; (B) histology grade; (C) clinical stage; (D) pT status; (E) pN status; (F) ER status; (G) PR status; (H) Her2 status; (I) p53 status.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Expressing

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 4. PinX1 inhibits proliferation in a human basal-like breast cancer cell line. (A) Immunoblot analysis of PinX1 expression levels in the PinX1- overexpressed group (PinX1-pEGFP-N1), wild-type MDA-MB-231 (WT) and negative control (pEGFP-N1). GAPDH is a loading control. (B) Immunoblot analysis of PinX1 expression levels in the PinX1-knockout group (PinX1-/-) and the wild-type control (MDA-MB-231). (C and D) The Cell Counting kit-8 (CCK-8) assay was applied to detect the proliferation rates at specific periods (0, 24, 48 and 72 h). Overexpression of PinX1 inhibits the proliferation of MDA-MB-231 while PinX1 knockout promotes its proliferation. **P<0.01.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Western Blot, Expressing, Negative Control, Control, Knock-Out, Cell Counting, CCK-8 Assay, Over Expression

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 5. PinX1 suppresses the migration ability of MDA-MB-231 cells in vitro. (A) The migration ability of the PinX1-overexpressed and the control groups were tested by wound healing assay in a 36-h recovery time (top, magnification, x20). Bottom, statistical analysis of the migration rates, presented relative to the initial wound area. (B) The migration ability of the PinX1 knockout and the wild-type groups was tested by wound healing assay in a 36-h recovery time (top, magnifications, x10). Bottom, statistical analysis of the migration rates, presented relative to the initial wound area. (C and D) Transwell assay was also used to measure the migration ability of the different experimental groups (left, magnifications, x10). Right, statistical analyses of the migrated cell number per field. At least three independent experiments were performed (*p<0.05, two-tailed unpaired t-test) and the data expressed are mean ± SEM.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Migration, In Vitro, Control, Wound Healing Assay, Knock-Out, Transwell Assay, Two Tailed Test

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 6. Overexpression of PinX1 induces apoptosis of MDA-MB-231 cells in vitro. (A and B) Western blot analysis was used to measure the expression levels of cleaved caspase-3 in different experimental groups. GAPDH was a loading control. (C) Flow cytometric analysis (left) of the apoptotic rate of the PinX1- overexpressed group with its negative control and the PinX1 knockout group with its counterpart. Quantitative analyses of the apoptotic rate on the right. At least three independent experiments were preformed (*p<0.05, two-tailed unpaired t-test) and the data expressed are mean ± SEM.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Over Expression, In Vitro, Western Blot, Expressing, Control, Negative Control, Knock-Out, Two Tailed Test