Journal: Cancer Cell International

Article Title: Sestrin2 reduces cancer stemness via Wnt/β-catenin signaling in colorectal cancer

doi: 10.1186/s12935-022-02498-x

Figure Lengend Snippet: The effects of Sestrin2 on cancer stemness are mediated by the Wnt/β-catenin pathway. A Western blot analysis of Wnt/β-catenin signaling-related proteins. The relative protein expression is shown in the right panel. β-Actin was used as a loading control * P = 0.05; Mann–Whitney test; lines showed medians). B Sphersphere formation assay in LV-Sestrin2 and LV-GFP CRC cells with or without exposure to BML-284 (0.1 µM). C Number of spheres, * compared with LV-GFP, * P = 0.05; # compared with LV-Sestrin2, # P = 0.05; Mann–Whitney test; lines showed medians. D Western blot analysis of proteins related to the Wnt/β-catenin pathway (β-catenin, c-Myc) and cancer stemness (CD44) in LV-Sestrin2 and LV-GFP CRC cells with or without incubation with BML-284 (0.5 µM) for 24 h in HCT-116 cells. E The relative protein expression. * Compared with LV-GFP, * P = 0.05, # compared with LV-Sestrin2, # P = 0.05, Mann–Whitney test; lines showed medians

Article Snippet: HCT-116 cells in the LV-Sestrin2 and LV-GFP groups were treated with 0.5 µM BML-284 (Wnt signaling activator; MedChemExpress, Monmouth Junction, NJ, USA) in dimethyl sulfoxide for 24 h and then collected for western blot analysis.

Techniques: Western Blot, Expressing, MANN-WHITNEY, Tube Formation Assay, Incubation

Journal: Cell Death & Disease

Article Title: Oncogene PRR14 promotes breast cancer through activation of PI3K signal pathway and inhibition of CHEK2 pathway

doi: 10.1038/s41419-020-2640-8

Figure Lengend Snippet: CHEK2 mRNA expression ( a ), CHEK2 protein expression ( b ) and p-CHEK2 (T68) protein expression ( c ) in PRR14 genetically unaltered and altered (amplified and mutated) breast cancer cases in TCGA database are statistically analyzed by two-tailed Student’s t -test. CHEK2 protein expression is detected by immunostaining in xenografts in nude mice from established MCF7 cell lines and MDA-MB-231 cell lines ( d ), as well as human breast cancer ( e ). CHEK2 transcription in human breast cancer is also detected by qRT-PCR ( f ). The data are quantified and two-tailed Student’s t -test is employed to determine the significance of the difference. Established MCF7 and MDA-MB-231 ( g ) PRR14-overexpressing and control cell lines are treated with various of genotoxic chemicals including Bleo, Eto, 5-FU, H2O2 and HU at indicated concentrations for indicated time. Key components of the ATM/CHEK2/P53 signaling pathway are detected by immunostaining. And CHEK2 protein expression ( h ) and mRNA expression ( i ) are detected by immunostaining and qRT-PCR, respectively. The data are analyzed by two-tailed Student’s t -test. Established MCF7 and MDA-MB-231 ( j ) PRR14-overexpressing and control cell lines are treated with Eto at indicated concentration for indicated time to induce p-CHEK2 (T68), which is detected by immunostaining and quantified and normalized by CHEK2 total protein ( k ). The data are analyzed by two-tailed Student’s t -test.

Article Snippet: Chemicals including selective CHEK2 inhibitor BML-277 (BML, HY-13946, MCE) , and genotoxic chemicals, including bleomycin (Bleo, HY-17565, MCE), etoposide (Eto, HY-13629, MCE), 5-fluorouracil (5-FU, HY-90006, MCE), H 2 O 2 (88597, Millipore) and hydroxyurea (HU, HY-B0313, MCE), were used to treat cells.

Techniques: Expressing, Amplification, Two Tailed Test, Immunostaining, Quantitative RT-PCR, Concentration Assay

Journal: Cell Death & Disease

Article Title: Oncogene PRR14 promotes breast cancer through activation of PI3K signal pathway and inhibition of CHEK2 pathway

doi: 10.1038/s41419-020-2640-8

Figure Lengend Snippet: MCF7 cells transfected with indicated siRNA sequences for 48 h are treated w/wo 5 μg/ml Eto for 6 h, then cells are harvested for immunostaining ( a ). PRR14 in cells is depleted by RNAi transfection for 48 h, and CHEK2 mRNA level are quantified by qRT-PCR in both MCF7 and MDA-MB-231 cell lines ( b ). MCF7, 10AKRAS, 7E6 and MDA-MB-231 cell lines are treated with increasing concentration of CHEK2 inhibitor BML for 24 h and followed with a 24 h treatment of Eto at 5 μg/ml. Cells are harvested to stain with PI followed with FACS analysis ( c ). The percentage of the 4N fraction is analyzed by one-way ANOVA analysis ( n = 3) ( d ).

Article Snippet: Chemicals including selective CHEK2 inhibitor BML-277 (BML, HY-13946, MCE) , and genotoxic chemicals, including bleomycin (Bleo, HY-17565, MCE), etoposide (Eto, HY-13629, MCE), 5-fluorouracil (5-FU, HY-90006, MCE), H 2 O 2 (88597, Millipore) and hydroxyurea (HU, HY-B0313, MCE), were used to treat cells.

Techniques: Transfection, Immunostaining, Quantitative RT-PCR, Concentration Assay, Staining

Journal: Cell Death & Disease

Article Title: Oncogene PRR14 promotes breast cancer through activation of PI3K signal pathway and inhibition of CHEK2 pathway

doi: 10.1038/s41419-020-2640-8

Figure Lengend Snippet: KM survival curves of breast cancer patients receiving chemotherapy are stratified by their expression levels of either CHEK2 ( a ) or PRR14 ( b ). For comparison, KM survival curves of breast cancer patients receiving endocrine therapy and stratified by PRR14 expression is also analyzed ( c ). KM survival curves of breast cancer patients receiving chemotherapy with mutant P53 ( d ) or wild-type P53 ( e ) are stratified by their expression level of PRR14.

Article Snippet: Chemicals including selective CHEK2 inhibitor BML-277 (BML, HY-13946, MCE) , and genotoxic chemicals, including bleomycin (Bleo, HY-17565, MCE), etoposide (Eto, HY-13629, MCE), 5-fluorouracil (5-FU, HY-90006, MCE), H 2 O 2 (88597, Millipore) and hydroxyurea (HU, HY-B0313, MCE), were used to treat cells.

Techniques: Expressing, Mutagenesis