Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Selectivity Index of Danazol on several multidrug-resistant cancer cell lines.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques:

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Cytotoxicity of natural and synthetic steroid hormones on HeLaS3 (parental) and KB/VIN (MDR) cancer cell lines.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques:

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Results of ABCB1 gene expression analysis. HeLaS3 and KB/VIN were treated with danazol for 24, 48, and 72 hours. Total RNA was extracted and ABCB1 gene expression level of each sample was quantified by real-time PCR. The ABCB1 gene expression was significantly down-regulated by danazol treatment in KB/VIN cell line. Statistical differences were evaluated by ANOVA followed post hoc analysis (Tukey’s test). * Indicates p value < 0.05 compared with control group. Data presented as mean ± SE of at least three experiments, each in triplicate.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Control

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Results of cell cycle analysis in HeLaS3 and MDR KB/VIN cells. HeLaS3 and KB/VIN were treated with culture medium ( a ), DMSO ( b ), paclitaxel ( c ; positive control), danazol ( d ), β-estradiol ( e ), and deoxycorticosterone ( f ) for 24, 48, and 72 hours. DNA contents and cell cycle distribution of each sample were determined by PI solution (X-axis PE). Danazol, β-estradiol, and deoxycorticosterone arrested KB/VIN cell at G2/M phase and caused apoptosis (increased sub G1) in a time-dependent manner.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Cell Cycle Assay, Positive Control

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: Results of caspase activity and ROS levels detection assay in HeLaS3 and MDR KB/VIN cells. HeLaS3 and KB/VIN cells were treated with or without danazol for 72 hours. The activities of caspase-8 ( a ) and caspase-9 ( b ) were evaluated by Cell Meter™ Caspase Activity Apoptosis Assay Kits. Danazol activated caspase-8 in KB/VIN cells while the level remained the same in HeLaS3 cells compared with respective control group. ( c ) The treatment of danazol increased the ROS level in both HeLaS3 and KB/VIN cell lines. Menadione, a ROS inducer, was used as a positive control. Statistical differences were evaluated by the ANOVA followed post hoc analysis (Tukey’s test). * indicates p value < 0.05 compared with control group. Data presented as mean ± SE of at least two experiments, each in duplicate.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Activity Assay, Detection Assay, Apoptosis Assay, Control, Positive Control

Journal: Scientific Reports

Article Title: Danazol mediates collateral sensitivity via STAT3/Myc related pathway in multidrug-resistant cancer cells

doi: 10.1038/s41598-019-48169-2

Figure Lengend Snippet: MYC signaling mediates danazol-induced G2/M arrest in MDR KB/VIN cells. HeLaS3 and KB/VIN were treated with or without danazol for 72 hours. The whole-treated cells were extracted and analyzed by ELISA using antibodies specific to human ( a ) STAT3-pY705, ( b ) cMYC, ( c ) CDC25, ( d ) CDK1, and ( e ) p21. The results showed danazol reduced the concentration of MYC (c-Myc) and its upstream protein STAT3-pY705 in MDR KB/VIN cells. In addition, decreases in downstream signaling proteins, CDC25 and CDK1, were shown. Each experiment was performed in duplicate. Student’s t-test, *p < 0.05 compared with cells treated with medium only control.

Article Snippet: Parental cancer cell lines, including human cervical epithelioid carcinoma HeLaS3, human hepatoblastoma HepG2, and human non-small cell lung carcinoma NCI-H460, were purchased from Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Control

Journal: The Journal of biological chemistry

Article Title: c-Myb trans-activates the human DNA topoisomerase IIalpha gene promoter.

doi: 10.1074/jbc.272.10.6278

Figure Lengend Snippet: FIG. 6. A dominant negative c-Myb peptide abrogates endoge- nous and exogenous c-Myb-mediated trans-activation of the hu- man topo IIa promoter. A, HL-60 cells were co-transfected with 2562TOP2LUC (20 mg), CMV-b-gal reference plasmid (1 mg), and 1 mg of either the empty CMV vector, CMV-myb, or CMV-mybDNRD. Sam- ples in each group also received either 1 mg of CMV vector, 1 mg of pSCDMS/MenT (a CMV-driven expression vector encoding the c-Myb DBD fused to the D. engrailed transcriptional repressor (34)) or 5 mg of pCMV-mybDBD (a passive dominant negative effector encoding the c-Myb DBD alone (34)). Cells were harvested at 24 h and processed for measurement of each reporter enzyme activity. Data is represented as -fold luciferase activity relative to the basal reporter activity in the absence of any effector plasmids. B, HL-60 and HeLa cells were co- transfected with 2562TOP2LUC (20 mg), CMV-b-gal reference plasmid (1 mg), and the indicated amounts of pSCDMS/MenT. Controls received 1 mg (HL-60) or 2 mg (HeLa) of empty CMV vector. Cells were processed for measurement of corrected reporter activity as in panel A.

Article Snippet: Cell Culture—HL-60 human promyelocytic leukemia cells (ATCC CCL 240), U937 histiocytic leukemia cells (ATCC CRL 1593), CCRFCEM lymphoblastic leukemia cells (ATCC 119), and HeLa S3 human cervical carcinoma cells (ATCC CCL 2.2) were obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Dominant Negative Mutation, Activation Assay, Transfection, Plasmid Preparation, Expressing, Activity Assay, Luciferase

Journal: The Journal of biological chemistry

Article Title: c-Myb trans-activates the human DNA topoisomerase IIalpha gene promoter.

doi: 10.1074/jbc.272.10.6278

Figure Lengend Snippet: FIG. 7. c-Myb trans-activation of 2562TOP2LUC in leukemia and epithelial cell lines. The T-cell leukemia (CCRF-CEM), promonocytic leukemia (U937), and cervical epithelial carcinoma (HeLa) lines were independently co-transfected with 2562TOP2LUC (20 mg), CMV-b-gal reference plasmid (1 mg), and the indicated amounts of CMV-myb expression plasmid. The control group in each case (0 mg of CMV-myb) received 2 mg of the empty CMV vector. Cells were harvested at 24 h and processed for measurement of each reporter enzyme activity. Numbers above each bar represent the -fold increase over each respective CMV only control.

Article Snippet: Cell Culture—HL-60 human promyelocytic leukemia cells (ATCC CCL 240), U937 histiocytic leukemia cells (ATCC CRL 1593), CCRFCEM lymphoblastic leukemia cells (ATCC 119), and HeLa S3 human cervical carcinoma cells (ATCC CCL 2.2) were obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Expressing, Control, Activity Assay

Journal: The Journal of biological chemistry

Article Title: c-Myb trans-activates the human DNA topoisomerase IIalpha gene promoter.

doi: 10.1074/jbc.272.10.6278

Figure Lengend Snippet: FIG. 8. Topo IIa promoter trans-activation by B-Myb. HL-60 and HeLa cells were independently co-transfected with 2562TOP2LUC (20 mg), RSV-b-gal reference plasmid (1 mg), and the indicated amounts of a CMV-B-myb expression plasmid (pKCB-myb). A b-gal internal control driven by the RSV promoter was required since the standard CMV-b-gal control was activated by B-Myb. The control group in each case received 15 mg of the empty CMV vector and each B-Myb group received an amount of CMV vector to bring the total to 15 mg. Cells were harvested at 24 h and processed for measurement of each reporter enzyme activity. Numbers above each bar represent the -fold increase over each respective CMV only control.

Article Snippet: Cell Culture—HL-60 human promyelocytic leukemia cells (ATCC CCL 240), U937 histiocytic leukemia cells (ATCC CRL 1593), CCRFCEM lymphoblastic leukemia cells (ATCC 119), and HeLa S3 human cervical carcinoma cells (ATCC CCL 2.2) were obtained from the American Type Culture Collection (Rockville, MD).

Techniques: Activation Assay, Transfection, Plasmid Preparation, Expressing, Control, Activity Assay

Journal: Cell Metabolism

Article Title: Ribosome stalling is a signal for metabolic regulation by the ribotoxic stress response

doi: 10.1016/j.cmet.2022.10.011

Figure Lengend Snippet:

Article Snippet: human cervical cancer S3 (HeLa S3) , ATCC , CRL-4000; RRID: CVCL 4388.

Techniques: Recombinant, Reverse Transcription, Staining, Clone Assay, Mutagenesis, SYBR Green Assay, DNA Extraction, Enzyme-linked Immunosorbent Assay, RNA Sequencing, CRISPR, Knock-Out, Quantitative RT-PCR, Software