Journal: Cell reports

Article Title: A multiplexed in vivo approach to identify driver genes in small cell lung cancer

doi: 10.1016/j.celrep.2023.111990

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human: NCI-H146 , ATCC , HTB-173; RRID:CVCL_1473.

Techniques: Plasmid Preparation, Virus, Recombinant, Modification, Labeling, Amplification, Polymer, Staining, Protease Inhibitor, Bicinchoninic Acid Protein Assay, Simple Western, Membrane, DNA Extraction, Purification, Sequencing, Generated, Software

Journal: International Journal of Molecular Sciences

Article Title: Impact of AHR Ligand TCDD on Human Embryonic Stem Cells and Early Differentiation

doi: 10.3390/ijms21239052

Figure Lengend Snippet: Expression of AHR and pluripotency marker genes in hESCs and embryoid bodies (EBs) cultured for 5 days. ( a ) Representative Western blot and densitometry analysis of AHR and actin protein levels in H9 cells. qPCR analysis of AHR ( b ), OCT4 , SOX2 and NANOG ( c ) mRNA levels in H1 and H9 hESCs and EBs. Data are presented relative to hESC H9 as means ± SEM from three independent experiments. * p < 0.05; ** p < 0.01.

Article Snippet: Human ES cell lines H1 (46, XY, WA01) and H9 (46, XX, WA09) were obtained from WiCell Research Institute (National Stem Cell Bank, Madison, WI, USA).

Techniques: Expressing, Marker, Cell Culture, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Impact of AHR Ligand TCDD on Human Embryonic Stem Cells and Early Differentiation

doi: 10.3390/ijms21239052

Figure Lengend Snippet: Analysis of lineage-specific marker gene expression in DMSO or TCDD pre-treated H9 hESCs and during directed differentiation into neural, endo- and mesodermal lineages. qPCR analysis of neural marker genes PAX6 ( a ) and OTX2 ( b ), endodermal marker genes SOX17 ( c ) and GATA4 ( d ) and mesodermal marker genes T ( e ) , HAND1 ( f ) and GSC ( g ) mRNA levels. Data are presented relative to hESC DMSO ( a , b , e , g ) or d2 DMSO ( c , d , f ) as means ± SEM from three independent experiments. * p < 0.05 vs. hESC; ** p < 0.01 vs. hESC; # p < 0.05 vs. d2; ## p < 0.01 vs. d2.

Article Snippet: Human ES cell lines H1 (46, XY, WA01) and H9 (46, XX, WA09) were obtained from WiCell Research Institute (National Stem Cell Bank, Madison, WI, USA).

Techniques: Marker, Expressing

Journal: Journal of Medicinal Chemistry

Article Title: Design of Bcl-2 and Bcl-xL Inhibitors with Subnanomolar Binding Affinities Based Upon a New Scaffold

doi: 10.1021/jm300178u

Figure Lengend Snippet: Binding affinities of our designed compounds to Bcl-2 and Bcl-X L proteins in our FP-based assays and inhibition of cell growth in two cancer cell lines.

Article Snippet: Human small cell lung cancer cell lines H146 and H1417 were purchased from the American Type Culture Collection (ATCC) and were maintained in RPMI-1640 medium containing 10% FBS.

Techniques: Binding Assay, Inhibition

Journal: Journal of Medicinal Chemistry

Article Title: Design of Bcl-2 and Bcl-xL Inhibitors with Subnanomolar Binding Affinities Based Upon a New Scaffold

doi: 10.1021/jm300178u

Figure Lengend Snippet: Binding affinities of our designed compounds to Bcl-2 and Bcl-X L proteins in FP-based assays and inhibition of cell growth in two small-cell lung cancer cell lines.

Article Snippet: Human small cell lung cancer cell lines H146 and H1417 were purchased from the American Type Culture Collection (ATCC) and were maintained in RPMI-1640 medium containing 10% FBS.

Techniques: Binding Assay, Inhibition

Journal: Journal of Medicinal Chemistry

Article Title: Design of Bcl-2 and Bcl-xL Inhibitors with Subnanomolar Binding Affinities Based Upon a New Scaffold

doi: 10.1021/jm300178u

Figure Lengend Snippet: (A). Cell death induction by 20 and 21 in the H146 cancer cell line. Cells were treated for 24 h and cell death was analyzed by trypan blue assay. (B). Induction of cleavage of PARP and caspase-3 by 20 and 21 in the H146 cell line. Cells were treated for 24 h and caspase-3 and PARP were probed by western blotting.

Article Snippet: Human small cell lung cancer cell lines H146 and H1417 were purchased from the American Type Culture Collection (ATCC) and were maintained in RPMI-1640 medium containing 10% FBS.

Techniques: Western Blot

Journal: BMC Cancer

Article Title: Vorinostat enhances the cisplatin-mediated anticancer effects in small cell lung cancer cells

doi: 10.1186/s12885-016-2888-7

Figure Lengend Snippet: Effects of triple combination treatments of vorinostat with cisplatin and etoposide on the viability and apoptosis of SCLC cells. H209 and H146 cells were treated with or without vorinostat in combination with cisplatin ( a vorinostat at 0.8 μM, and cisplatin and etoposide both at 1 μM; b vorinostat at 0.4 μM, cisplatin at 0.2 μM, and etoposide at 0.6 μM) for 24 h. Cell viability was determined using the MTS assay, and data were represented as mean ± SD in triplicate. A significant reduction in cell viability was documented (*, P < 0.05; **, P < 0.01; ***, P < 0.001) compared with vorinostat or cisplatin/etoposide alone. c , d PARP cleavage was used for determining apoptosis. The cells were treated with the same triple combination treatment described previously for 24 h, and cell lysates were collected and subjected to Western blot analysis with PARP and α-tubulin

Article Snippet: Human SCLC cell lines (H209 and H146) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: MTS Assay, Western Blot

Journal: BMC Cancer

Article Title: Vorinostat enhances the cisplatin-mediated anticancer effects in small cell lung cancer cells

doi: 10.1186/s12885-016-2888-7

Figure Lengend Snippet: Enhanced antigrowth activity of vorinostat combined with cisplatin in SCLC cells. a H209 and b H146 cells were treated with various concentrations of vorinostat (0.1 and 0.5 μM and 0.1 and 0.2 μM, respectively), cisplatin alone (5 μM), and combinations of the 2 drugs for 24 h. Cell viability was assessed using the MTS assay, and data were represented as mean ± SD in triplicate. A significant reduction in cell viability was documented (*, P < 0.05; ***, P < 0.001) compared with that in vorinostat or cisplatin alone

Article Snippet: Human SCLC cell lines (H209 and H146) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Activity Assay, MTS Assay

Journal: BMC Cancer

Article Title: Vorinostat enhances the cisplatin-mediated anticancer effects in small cell lung cancer cells

doi: 10.1186/s12885-016-2888-7

Figure Lengend Snippet: Vorinostat plus cisplatin triggers apoptotic cell death in SCLC cells. a H209 and b H146 cells were exposed to vorinostat or cisplatin alone and in combination with these 2 agents for 24 h. Cell lysates were collected and subjected to Western blot analysis with PARP and α-tubulin. In the caspase-3 activity assay, cells were treated with vorinostat alone (0.1 μM), cisplatin alone (5 μM), and in combination for c 24 and 36 h (H209) or d 24 h (H146). DEVDase activity was represented as mean ± SD in 3 independent studies. A significant induction of DEVDase activity was documented (*, P < 0.05; **, P < 0.01; *** P < 0.001) compared with that in vorinostat alone, cisplatin alone, or DMSO control

Article Snippet: Human SCLC cell lines (H209 and H146) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Western Blot, Caspase-3 Activity Assay, Activity Assay, Control

Journal: BMC Cancer

Article Title: Vorinostat enhances the cisplatin-mediated anticancer effects in small cell lung cancer cells

doi: 10.1186/s12885-016-2888-7

Figure Lengend Snippet: Cell cycle progression analysis of vorinostat in combination with cisplatin in SCLC cells. a H209 and b H146 cells were treated with vorinostat (0.1 μM), cisplatin alone (5 μM), and a combination of these 2 agents for 24 h, and the cell cycle distribution was analyzed through flow cytometry. c , d Statistical analysis of cell cycle distribution was represented as mean ± SD in 3 independent studies. A significant arrest in the cell-cycle phase was documented (*, P < 0.05; **, P < 0.01) compared with that in DMSO

Article Snippet: Human SCLC cell lines (H209 and H146) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Flow Cytometry

Journal: BMC Cancer

Article Title: Vorinostat enhances the cisplatin-mediated anticancer effects in small cell lung cancer cells

doi: 10.1186/s12885-016-2888-7

Figure Lengend Snippet: Effects of cotreatments on vorinostat-regulated signaling molecules in SCLC cells. a H209 and b H146 cells were treated with indicated concentrations of vorinostat (0.1, 0.25, and 0.5 μM and 0.05, 0.1, and 0.2 μM, respectively), cisplatin alone (5 μM), and a combination of these 2 agents for 24 h. Cell lysates were collected and subjected to Western blot analysis with acetyl α-tubulin, acetyl histone H3, TS, and β-actin

Article Snippet: Human SCLC cell lines (H209 and H146) were obtained from the Bioresource Collection and Research Center (Hsinchu, Taiwan).

Techniques: Western Blot

Journal: Cancer Medicine

Article Title: Poly (ADP) ribose polymerase enzyme inhibitor, veliparib, potentiates chemotherapy and radiation in vitro and in vivo in small cell lung cancer

doi: 10.1002/cam4.317

Figure Lengend Snippet: (A) Veliparib showed limited single-agent activity across a wide concentration range in a panel of SCLC cell lines. (B) Log of mean ± SEM of IC 50 concentrations for cisplatin, carboplatin, and etoposide alone and in combination with 5 and 50 μ mol/L concentrations of veliparib. Each bar represents log of the mean value obtained from 3 to 4 independent experiments. Bottom Right: Potentiation of cytotoxicity induced by gamma radiation in the presence of veliparib (5 μ mol/L) in 2 representative cell lines (H146 and DMS153).

Article Snippet: Human SCLC cell lines (H146, H187, H128, H69, H209, DMS153, H526, DMS114, and DMS53) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Activity Assay, Concentration Assay

Journal: Cancer Medicine

Article Title: Poly (ADP) ribose polymerase enzyme inhibitor, veliparib, potentiates chemotherapy and radiation in vitro and in vivo in small cell lung cancer

doi: 10.1002/cam4.317

Figure Lengend Snippet: (A) Variable expression of BRCA1, ERCC1, and DNA-PKcs across the panel of SCLC cell lines. DNA-PKcs expression was noticeably high in cell lines (H69, H128 and DMS53) that were less sensitive to cisplatin or PARP inhibitor potentiation of cisplatin activity. (B) Top panel: Reduced expression of DNA-PKcs observed in H146 cell line coincided with onset of apoptosis as indicated by caspase 3 cleavage. Bottom Panel: DNA-PKcs expression was modulated in H146 and H128 cell lines but only at high concentrations of cisplatin and veliparib required for cytotoxicity (see bar graphs showing normalized DNA-PKcs expression relative to actin in H146 and H128 cell lines under different treatment conditions, 1–9, as measured by densitometry). There was no significant modulation of other representative DNA-repair enzymes. (C) Inhibition of PARP activity observed within 1 h of cell exposure to veliparib and persisted for the duration of the experiments 24 h later. Reduced DNA-PKcs expression observed at 24 h coincident with onset of apoptosis as indicated by cleaved caspase 3 and PARP (bar graphs showing normalized DNA-PKcs expression relative to actin in H146 cell lines as measured by densitometry under different treatment conditions, 1–9 and at different time points). Lanes: 1, Control; 2, cisplatin (2.5 μ mol/L); 3, cisplatin (50 μ mol/L); 4, veliparib (5 μ mol/L); 5, veliparib (50 μ mol/L); 6, cisplatin (2.5 μ mol/L) + veliparib (5 μ mol/L); 7, cisplatin (2.5 μ mol/L) + veliparib (50 μ mol/L); 8, cisplatin (50 μ mol/L) + veliparib (5 μ mol/L); 9, cisplatin (50 μ mol/L) + veliparib (50 μ mol/L).

Article Snippet: Human SCLC cell lines (H146, H187, H128, H69, H209, DMS153, H526, DMS114, and DMS53) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Activity Assay, Inhibition, Control

Journal: Cancer Medicine

Article Title: Poly (ADP) ribose polymerase enzyme inhibitor, veliparib, potentiates chemotherapy and radiation in vitro and in vivo in small cell lung cancer

doi: 10.1002/cam4.317

Figure Lengend Snippet: (A) Genomic DNA and RNA extracted from untreated exponentially growing cells as described were employed for gene expression profiling on the Illumina and NanoString nCounter platforms. Unsupervised analysis of the native gene expression profiles of the 9 SCLC cell lines was conducted using principal component analysis (PCA) method. Five cell lines observed to cluster together (Red) were the same cell lines that displayed increased sensitivity to cisplatin and the potentiation of cisplatin by veliparib. (B) Cells were treated by continuous exposure for 24 h to vehicle, Con, control; A5, veliparib (5 μ mol/L); A50, veliparib (50 μ mol/L); C + 0, cisplatin alone (IC 50 concentration for the respective cell lines); C + 5, cisplatin + veliparib (5 μ mol/L); C + 50, cisplatin + veliparib (50 μ mol/L); Rad, radiation (single time point exposure to 2 Gy). Harvested cells were employed for DNA and RNA extraction as described and employed for gene expression profiling on the Illumina and NanoString nCounter platforms. PCA of the gene expression profiles of SCLC cell lines under different treatment conditions. Cells were observed to cluster principally by cell of origin rather than by specific type of treatment. (C) Venn diagram showing the distribution of the 23 differentially expressed genes and pseudo genes when untreated and treated cell lines were compared based on the sensitivity to cisplatin and PARP inhibitor potentiation of cisplatin cytotoxicity. (D) Left: Cluster analysis of Illumina expression data using the median expression value as cut off for the genes of interest showed clustering of genes according to sensitive (blue) and insensitive (red) status. Right: Cluster analysis of NanoString nCounter expression data showed separation of the cell lines into two groups, sensitive (blue) and insensitive (red).

Article Snippet: Human SCLC cell lines (H146, H187, H128, H69, H209, DMS153, H526, DMS114, and DMS53) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Gene Expression, Control, Concentration Assay, RNA Extraction, Expressing

Journal: Endocrinology

Article Title: Blockade of Sphingosine 1-Phosphate Receptor 2 Signaling Attenuates High-Fat Diet-Induced Adipocyte Hypertrophy and Systemic Glucose Intolerance in Mice

doi: 10.1210/en.2015-1768

Figure Lengend Snippet: Antibody Table

Article Snippet: Leptin , A recombinant protein coding amino acids 22–167 of human Ob , Ob (H-146) , Santa Cruz Biotechnology, Inc, sc-9014 , Rabbit; polyclonal , 50 (WB[Millipore SNAP i.d]) , 10.1074/jbc.270.46.27728.

Techniques: Sequencing, Expressing, Plasmid Preparation, Residue, Phospho-proteomics, Derivative Assay, Recombinant