Journal: Cell Death Discovery

Article Title: Repositioning of a cyclin-dependent kinase inhibitor GW8510 as a ribonucleotide reductase M2 inhibitor to treat human colorectal cancer

doi: 10.1038/cddiscovery.2016.27

Figure Lengend Snippet: Role of RRM2 in CRC. ( a ) Summary view of RRM1, RRM2, and RRM2B expression profiles in human tumors using published human oncology microarray data (Oncomine). The number in each cell under ‘Cancer versus Normal’ corresponds to the amount of cancer types that contains a significantly different level of RRM1, RRM2, or RRM2B compared with normal corresponding tissue. Thresholds for significance are: fold expression>2; P -value<0.05 and ranking of gene in the analyses>top 10%. Red signifies the gene overexpression in the analyses; blue represents the gene underexpression. Intensity of color signifies the best rank of gene in those analyses. ( b ) A microarray dataset (GSE8671) of colorectal adenomas and adjacent normal mucosa was obtained from NCBI GEO database. Probe IDs of RRM2: 209773_s_at. ( c ) A microarray dataset (GSE1710) of patients with Crohn’s disease and ulcerative colitis was obtained from NCBI GEO database. RRM2 expression in these datasets was shown.

Article Snippet: Horseradish peroxidase-labeled goat anti-rabbit and anti-mouse secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA). pcDNA3-RRM2 plasmid was purchased from Addgene (Cambridge, MA, USA).

Techniques: Expressing, Microarray, Over Expression

Journal: Cell Death Discovery

Article Title: Repositioning of a cyclin-dependent kinase inhibitor GW8510 as a ribonucleotide reductase M2 inhibitor to treat human colorectal cancer

doi: 10.1038/cddiscovery.2016.27

Figure Lengend Snippet: Identification of GW8510 as a potential RRM2 inhibitor. ( a ) The chemical structures of GW8510. ( b ) HCT116 cells were treated with various doses of GW8510 for 72 h. The cell viability was analyzed by an MTT assay. ( c ) HCT116 cells were treated with various doses of GW8510 for 24 h. The protein expressions were analyzed by western blots. ( d ) HCT116 cells were treated with various doses of GW8510 for 24 h in the absence or presence of 5 μ M MG132. The protein expressions were analyzed by western blots. ( e ) HCT116 cells were transiently transfected with a RRM2-overexpressing (pcDNA3-RRM2) or a control (pcDNA3) plasmid for 48 h, and then treated with indicated doses of GW8510 for 24 h. The protein expressions were analyzed by western blots. ( f ) HCT116 cells were transiently transfected with a RRM2-overexpressing (pcDNA3-RRM2) or a control (pcDNA3) plasmid for 24 h, and then treated with indicated doses of GW8510 for 72 h. The cell viability was analyzed by an MTT assay.

Article Snippet: Horseradish peroxidase-labeled goat anti-rabbit and anti-mouse secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA). pcDNA3-RRM2 plasmid was purchased from Addgene (Cambridge, MA, USA).

Techniques: MTT Assay, Western Blot, Transfection, Control, Plasmid Preparation

Journal: Cell Death Discovery

Article Title: Repositioning of a cyclin-dependent kinase inhibitor GW8510 as a ribonucleotide reductase M2 inhibitor to treat human colorectal cancer

doi: 10.1038/cddiscovery.2016.27

Figure Lengend Snippet: The gene expression signatures of compounds most positively (mean score> 0.7 and P <0.01) correlated with that of RRM2 siRNA

Article Snippet: Horseradish peroxidase-labeled goat anti-rabbit and anti-mouse secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA). pcDNA3-RRM2 plasmid was purchased from Addgene (Cambridge, MA, USA).

Techniques: Gene Expression, Activity Assay

Journal: Cell Death Discovery

Article Title: Repositioning of a cyclin-dependent kinase inhibitor GW8510 as a ribonucleotide reductase M2 inhibitor to treat human colorectal cancer

doi: 10.1038/cddiscovery.2016.27

Figure Lengend Snippet: GW8510 induced autophagic cell death of CRC cells. ( a ) HCT116 cells were treated with various doses of GW8510 for 24 h. The protein expressions were analyzed by western blots. ( b ) HCT116 cells were treated with various doses of GW8510 for 48 h. The protein expressions were analyzed by western blots. ( c ) HCT116 cells were treated with 2 μ M GW8510 for 48 h in the absence or presence of 50 μ M ZVAD-FMK. The protein expressions were analyzed by western blots. ( d ) HCT116 cells were transiently transfected with RRM2 siRNA for 48 h, and then treated with 4 μ M GW8510 for 24 h. The protein expressions were analyzed by western blots. ( e ) HCT116 cells were transiently transfected with a RRM2-overexpressing (pcDNA3-RRM2) or a control (pcDNA3) plasmid for 48 h, and then treated with 4 μ M GW8510 for 24 h. The protein expressions were analyzed by western blots.

Article Snippet: Horseradish peroxidase-labeled goat anti-rabbit and anti-mouse secondary antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA). pcDNA3-RRM2 plasmid was purchased from Addgene (Cambridge, MA, USA).

Techniques: Western Blot, Transfection, Control, Plasmid Preparation

Journal: eLife

Article Title: Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS

doi: 10.7554/eLife.27574

Figure Lengend Snippet:

Article Snippet: Antibody , rabbit anti-RRM2 , Human Protein Atlas , RRID: AB_2683304 , .

Techniques: Expressing, Construct