Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: Identification of signaling pathways and genes related to PTX resistance in breast cancer cell lines. (A) Venn diagram distribution of differentially expressed genes from 2 arrays. As revealed, 518 genes were specifically differentially expressed in GSE28784 cells and 1,778 genes were specifically differentially expressed in GSE90564 cells, while 151 genes were expressed in both datasets. (B) GO annotations and a functional pathway analysis of 151 genes performed using GO-terms and KEGG. (C) Gene-set enrichment analysis of common differentially regulated genes from A. (D) Gene interaction network constructed based on the shared gene list from A. Different colors represent different clusters as calculated using the K-means clustering method. Core genes in the network were identified based on their edge connectivity number. (E) Gene interaction map related to SERPINE1 generated using STRING. PTX, paclitaxel; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and genomes; SERPINE1 , serine protease inhibitor, clade E member 1; STRING, Search Tool for the Retrieval of Interacting Genes.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Protein-Protein interactions, Functional Assay, Construct, Generated, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: SERPINE1 is upregulated in PTX-resistant BC cell lines. (A) Survival curves for MDA-MB-231 PTX-resistant and -sensitive cells treated with various concentrations of PTX. *P<0.05, **P<0.01. (B) Survival curves for MCF-7 PTX-resistant and -sensitive cells treated with various concentrations of PTX. **P<0.01, ***P<0.001. Some points have no error bars because the standard deviation is too small in A and B. (C) SERPINE1 mRNA levels in MDA-MB-231 PTX-resistant vs. -sensitive cells as assessed by RT-qPCR. ***P<0.001. (D) SERPINE1 mRNA levels in MCF-7 PTX-resistant vs. -sensitive cells as assessed by RT-qPCR. ***P<0.001. (E) SERPINE1 protein expression in MDA-MB-231 and MCF-7 PTX-resistant vs. -sensitive cells as measured by western blotting. **P<0.01. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Standard Deviation, Quantitative RT-PCR, Expressing, Western Blot, Protease Inhibitor, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: Knockdown of SERPINE1 suppresses PTX-resistant BC cell survival and promotes apoptosis. (A) Western blots revealing the levels of SERPINE1 protein in knockdown cells. **P<0.01. (B) Survival curves for MDA-MB-231 and MCF-7 PTX-resistant cells with/without SERPINE1 suppression and treated with various concentrations of PTX. *P<0.05, **P<0.01. Some points have no error bars because the standard deviation is too small. (C) Flow cytometric detection of cell apoptosis among BC cells with/without SERPINE1 suppression. *P<0.05, ***P<0.001. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Knockdown, Western Blot, Standard Deviation, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: SERPINE1 suppression induces apoptosis in PTX-resistant TNBC cells. (A) Levels of cleaved caspase-3 and Bax proteins in the control and SERPINE1 -knockdown MDA-MB-231 PTX-resistant cells. (B) Levels of cleaved caspase-3 and Bax proteins in the control and SERPINE1 -knockdown MCF-7 PTX-resistant cells. (C) Quantification of cleaved caspase-3 protein levels in the control and SERPINE1 -knockdown PTX-resistant cells. **P<0.01. (D) Quantification of Bax protein levels in the control and SERPINE-knockdown PTX-resistant cells. **P<0.01. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; TNBC, triple-negative breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Control, Knockdown, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: Downregulation of SERPINE1 decreases the tumorigenic capacity of PTX-resistant BC cells in vivo . (A) Image revealing the appearance of tumors from mice that were inoculated with MDA-MB-231/PTX cells with or without SERPINE1 knockdown. (B) Quantification of tumor volumes at the end of the in vivo tumorigenic assays. **P<0.01. SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: In Vivo, Knockdown, Protease Inhibitor

Journal: Oncology Reports

Article Title: Knockdown of SERPINE1 reverses resistance of triple-negative breast cancer to paclitaxel via suppression of VEGFA

doi: 10.3892/or.2020.7770

Figure Lengend Snippet: VEGFA is regulated by SERPINE1 and mediates PTX resistance in BC cells. (A) Survival curves for MDA-MB-231 PTX-resistant cells with/without SERPINE1 suppression and supplemented with VEGFA . *P<0.05, **P<0.01. (B) Survival curves for MCF-7/PTX cells with/without SERPINE1 suppression and supplemented with VEGFA . Some points have no error bars because the standard deviation was too small in A and B. *P<0.05, **P<0.01. (C) Flow cytometric evaluation of apoptosis among BC cells with/without SERPINE1 suppression and treated with VEGFA . **P<0.01 vs. NC group; ## P<0.01 vs. sh- SERPINE1 group. (D) Expression of VEGFA protein in BC cells with/without SERPINE1 suppression and treated with VEGFA . **P<0.01. VEGFA , vascular endothelial growth factor A; SERPINE1 , serine protease inhibitor, clade E member 1; PTX, paclitaxel; BC, breast cancer.

Article Snippet: The membranes were blocked for 1 h with 5% skimmed milk at room temperature and incubated with anti-SERPINE1 antibody (cat. no. A00637-1; 1:1,000 dilution; Boster Biological Technology), VEGFA (cat. no. BA0407; 1:500 dilution; Wuhan Boster Biological Technology Co., Ltd.), cleaved caspase-3 (product code ab32042; 1:500 dilution; Abcam), Bax (cat. no. A00183; 1:1,000 dilution; Wuhan Boster Biological Technology Co., Ltd.) and β-actin (product code ab8224; 1:1,000 dilution; Abcam) overnight at 4°C.

Techniques: Standard Deviation, Expressing, Protease Inhibitor