Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Association between clinicopathological factors and the expression of β-catenin.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Expression of β-catenin in BC tissues and cell lines. The expression of β-catenin was determined in 32 paired BC tissues at the (A) mRNA and (B) protein levels were determined by RT-qPCR and western blot analysis, respectively. (C) The expression of β-catenin was analyzed in BC tissues by immunohistochemistry. Magnification, ×200. (D) Score analyses of the immunohistochemistry results (n=32 vs. 32). The expression levels of β-catenin in the BC MCF-10A, MDA-MB-468 and T47D cell lines and MCF-7 cells at the (E) mRNA and (F and G) protein levels were determined by RT-qPCR and western blot analysis, respectively. All data are presented as the mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. adjacent tissues or normal cells MCF-10A. BC, breast cancer; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Viability of BC cell lines and the expression of β-catenin are regulated by cisplatin and siRNA interference. The viability of (A) T47D and (B) MCF-7 cells was inhibited by cisplatin at different concentration (20, 40, 80 and 160 nM) determined by CCK-8 assays for 24 h. (C) MCF-7 cells were either not transfected or transfected with control siRNA or siR-β-catenin. At 24 h post transfection, cells were lysed and β-catenin expression was determined by western blot analysis. The viability of (D) T47D and (E) MCF-7 cells was suppressed by the combination of cisplatin (80 nM) and siR-β-catenin for 24 h. Each sample was analyzed in triplicate and was normalized to the control. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer; siRNA, small interfering RNA.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Expressing, Concentration Assay, CCK-8 Assay, Transfection, Western Blot, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Migration and invasion abilities of BC cells are suppressed by combined treatment with cisplatin and siR-β-catenin. The migratory abilities of (A and B) T47D cells and (C and D) MCF-7 cells treated with cisplatin and siR-β-catenin was evaluated using Transwell assays. The invasive ability of (E and F) T47D cells and (G and H) MCF-7 cells treated with cisplatin and siR-β-catenin was determined using Transwell assays. All data are presented as the mean ± standard error of the mean of 3 independent experiments. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer; siRNA, small interfering RNA.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Migration, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Expression levels of CD44/54 in BC cells are analyzed by flow cytometry. (A) T47D cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin were analyzed by FACS and the fluorescence intensities of CD44/CD54 were obtained. (B) Statistical analysis of the expression of CD44/CD54 in T47D cells. (C) The expression of CD44/CD54 in MCF-7 cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin were analyzed by FACS. (D) Statistical analysis of the expression of CD44/CD54 in MCF-7 cells. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer; siRNA; small interfering RNA; CD44, CD44 antigen; CD54, intercellular adhesion molecule 1.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Expressing, Flow Cytometry, Fluorescence, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Cell cycle distribution of BC cells treated with the combination of cisplatin and siR-β-catenin detected by flow cytometry. (A) The cell cycle distribution of T47D cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin was determined by flow cytometry. (B) Statistical analysis of the cell cycle analysis results of T47D cells. (C) The cell cycle distribution of MCF-7 cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin was determined by flow cytometry. (D) Statistical analysis of the cell cycle analysis results of T47D cells. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Flow Cytometry, Cell Cycle Assay

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Cisplatin- and siR-β-catenin-induced apoptosis of BC cells is measured using flow cytometry. (A) T47D cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin were examined by flow cytometry. (B) Statistical analysis of apoptosis assay results in T47D cells. (C) The levels of apoptosis in MCF-7 cells treated with cisplatin, siR-β-catenin and the combination of cisplatin and siR-β-catenin was determined by flow cytometry. (D) Statistical analysis of apoptosis assay results in MCF-7 cells. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer; siRNA, small interfering RNA.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Flow Cytometry, Apoptosis Assay, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Levels of apoptosis in BC cells induced by treatment with cisplatin and siR-β-catenin in combination are analyzed using Hoechst 33258 staining. (A and B) Apoptosis was significantly increased in T47D cells treated with the combination of cisplatin and siR-β-catenin. (C and D) Apoptosis was significantly increased in MCF-7 cells treated with the combination of cisplatin and siR-β-catenin. Nuclear morphological changes were observed under a fluorescence microscope. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001, vs. control. BC, breast cancer; siRNA, small interfering RNA.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Staining, Fluorescence, Microscopy, Small Interfering RNA

Journal: International Journal of Molecular Medicine

Article Title: Effects of cisplatin on the proliferation, invasion and apoptosis of breast cancer cells following β-catenin silencing

doi: 10.3892/ijmm.2020.4543

Figure Lengend Snippet: Proteins of the β-catenin signaling pathway and apoptosis-associated proteins are regulated by treatment with cisplatin and siR-β-catenin in combination. (A) The expression levels of signaling pathway proteins β-catenin, c-Myc and cyclin D1 were suppressed by the combination of cisplatin and siR-β-catenin in MCF-7 cells. (B) Statistical analysis of the expression levels of β-catenin, c-Myc and cyclin D1 in MCF-7 cells. (C) The levels of apoptosis-associated proteins caspase-3 and caspase-9 were increased by the treatment of combination of cisplatin and siR-β-catenin in MCF-7 cells. (D) Statistical analysis of caspase-3 and caspase-9 expression in MCF-7 cells. All data are presented as mean ± standard error of the mean. * P<0.05, ** P<0.01 and *** P<0.001 vs. control. BC, breast cancer; siRNA, small interfering RNA; c-Myc, MYC proto-oncogene, BHLH transcription factor.

Article Snippet: Once the cells reached 95% confluence, they were trans-fected with a SignalSilence ® β-catenin siRNA II (siR-β-catenin; cat. no. 6238; Cell Signaling Technology, Inc.) or unconjugated SignalSilence ® control siRNA (cat no. 6568; Cell Signaling Technology, Inc.) with Lipofectamine 2000™ (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's instructions.

Techniques: Expressing, Small Interfering RNA

Journal: Scientific Reports

Article Title: Polyphyllin I suppresses human osteosarcoma growth by inactivation of Wnt/β-catenin pathway in vitro and in vivo

doi: 10.1038/s41598-017-07194-9

Figure Lengend Snippet: PPI suppressed osteosarcoma cells by specifically inactivating Wnt/β-catenin signaling pathway. ( A ) RT-PCR analysis of β-catenin expression level in matched human osteosarcoma tissues (tumors) and adjacent noncancerous tissues (normal) from 3 patients. ( B ) 143-B cells and ( C ) HOS cells were respectively treated with 0.8 μM PPI for indicated times, and expressions of test proteins were examined by western blotting analysis, β-actin was used as loading control, and the full-length blots were included in the supplementary information file as Figures and . 143-B cells were pretreated with 4 μM CHIR9902 (the specific GSK-3β inhibitor) for 24 h before exposed to 0.8 μM PPI for another 48 h, the combined CHIR and PPI treatments result in rescued ( D ) active β-catenin expression (the full-length blots were included in the supplementary information file as Figure ) and cell viability ( E ) compared to the PPI treatment alone in 143-B osteosarcoma cells. 143-B cells were transfected with either small interfering RNA-targeting β-catenin (si-β-catenin) or si-control for 48 h before exposed to 0.8 μM PPI for another 24 h, si-β-catenin potentiated PPI induced ( F ) down-regulation of active β-catenin expression (the full-length blots were included in the supplementary information file as Figure ), ( G ) decrease of cell viability and ( H ) inhibition of migration of 143-B osteosarcoma cells induced by PPI. * p < 0.05, ** p < 0.01, *** p < 0.001, versus the control.

Article Snippet: Briefly, 143-B cells were co-transfected with either small interfering RNA-targeting β-catenin (100 nM si-β-catenin) or 100 nM si-control for 72 h, using Lipofectamine 2000 (Invitrogen). β-catenin siRNA II (#6238) was bought from Cell Signaling Technology (Danvers, USA). si-control was designed and produced by Shanghai GenePharma Co.,Ltd (Shanghai, China).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Transfection, Small Interfering RNA, Inhibition, Migration

Journal: Oncotarget

Article Title: Efficacy of glycogen synthase kinase-3β targeting against osteosarcoma via activation of β-catenin

doi: 10.18632/oncotarget.12781

Figure Lengend Snippet: ( A ) Western-blotting analysis was used to compare the expression and phosphorylation of β-catenin between cells treated with DMSO and either GSK-3β inhibitor. Expression of β-actin was monitored as a loading control. ( B ) The left panels show representative immunofluorescence microscopic findings of expression and subcellular localization of β-catenin in osteosarcoma (143B, MG-63) and osteoblast (hFOB1.19) cells. The scale bar in each panel indicates 25 μm. The number shown below each panel indicates the percentage of nuclear β-catenin-positive cells among the total number of cells. The bar graphs on the right shows the effects of DMSO and AR-A014418 on the incidence of nuclear localization of β-catenin in osteosarcoma and osteoblast cells. In each assay, the mean percentage of nuclear β-catenin-positive cells in 3 microscopic fields was evaluated with standard deviation. ( C ) Relative co-transcriptional activity of β-catenin was measured by the TOP/FOP flash assay and compared between cells treated with DMSO, AR-A014418 and SB-216763, respectively. (B, C) Asterisks denote a statistically-significant difference between the data after administration of vehicle and GSK-3β inhibitors.

Article Snippet: Osteosarcoma cells were transfected with 15 nmol/l non-specific (SignalSilence control siRNA, Cell Signaling Technology) or β-catenin-specific siRNA (SignalSilence β-catenin siRNA II, Cell Signaling Technology) for 18 hrs.

Techniques: Western Blot, Expressing, Immunofluorescence, Standard Deviation, Activity Assay

Journal: Oncotarget

Article Title: Efficacy of glycogen synthase kinase-3β targeting against osteosarcoma via activation of β-catenin

doi: 10.18632/oncotarget.12781

Figure Lengend Snippet: (A) Tumor size was measured weekly and the volume calculated. (B) Mean weight of the tumors removed at necropsy. Asterisks denote a statistically-significant difference in tumor volume and weight compared to mice treated with DMSO. The scatter plots corresponding to the data in (A) and (B) are shown in . ( C ) Histological and immunohistochemical findings for orthotopic tumors in mice treated with DMSO or with GSK-3β inhibitors. Representative paraffin-embedded sections of tumors were stained with H&E or immunostained for β-catenin. Magnified images of the sections (upper three panels) are shown in . Higher resolution versions of the lower six panels are shown in . Scale bar in each of the upper three panels indicates 5 mm and that of the middle and lower six panels indicates 100 μm.

Article Snippet: Osteosarcoma cells were transfected with 15 nmol/l non-specific (SignalSilence control siRNA, Cell Signaling Technology) or β-catenin-specific siRNA (SignalSilence β-catenin siRNA II, Cell Signaling Technology) for 18 hrs.

Techniques: Immunohistochemical staining, Staining