vulvar squamous cell carcinoma  (ATCC)

Journal: Cancer medicine

Article Title: Evaluation of CHK1 activation in vulvar squamous cell carcinoma and its potential as a therapeutic target in vitro.

doi: 10.1002/cam4.1638

Figure Lengend Snippet: FIGURE 1 Expression of pCHK1Ser345, pCHK1Ser317, pCHK1Ser296, and pCHK1Ser280 in vulvar squamous epithelium. Immunostaining of pCHK1Ser345 (A), pCHK1Ser317 (B), pCHK1Ser296 (C), and pCHK1Ser280 (D) in normal vulvar epithelium (Magnification × 300). High expression of pCHK1Ser345 (E), pCHK1Ser317 (F), pCHK1Ser296 (G), and pCHK1Ser280 (H) and low expression of pCHK1Ser345 (I), pCHK1Ser317 (J), pCHK1Ser296 (K), and pCHK1Ser280 (L) in VSCC (Magnification × 600)

Article Snippet: Vulvar squamous cell carcinoma cell line CAL39 (DSMZ, Braunschweig Germany) was cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, LifeTechnologies, Invitrogen, Oslo, Norway) supplemented with 10% Fetal Calf Serum (Biocrom, KG, Berlin, Germany) and 2 mM Lglutamine (Lonza, Vervieres, Belgium) at 37°C in humidified condition containing 5% CO2.

Techniques: Expressing, Immunostaining

Journal: Cancer medicine

Article Title: Evaluation of CHK1 activation in vulvar squamous cell carcinoma and its potential as a therapeutic target in vitro.

doi: 10.1002/cam4.1638

Figure Lengend Snippet: FIGURE 2 Expression of pCHK1Ser345, pCHK1Ser317, pCHK1Ser296, and pCHK1Ser280 in vulvar cancer cell lines. Immunostaining of pCHK1Ser345, pCHK1Ser317, pCHK1Ser296, and pCHK1Ser280 in SW945 (A-D) and CAL39 (E-H) (Magnification × 600)

Article Snippet: Vulvar squamous cell carcinoma cell line CAL39 (DSMZ, Braunschweig Germany) was cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, LifeTechnologies, Invitrogen, Oslo, Norway) supplemented with 10% Fetal Calf Serum (Biocrom, KG, Berlin, Germany) and 2 mM Lglutamine (Lonza, Vervieres, Belgium) at 37°C in humidified condition containing 5% CO2.

Techniques: Expressing, Immunostaining

Journal: Cancer medicine

Article Title: Evaluation of CHK1 activation in vulvar squamous cell carcinoma and its potential as a therapeutic target in vitro.

doi: 10.1002/cam4.1638

Figure Lengend Snippet: FIGURE 4 AZD7762 reduces viability in vulvar squamous cells. (A and C) Cell viability measured by CTG assay in CAL39 and SW954 exposed to the increasing concentrations of AZD7762 for 3 days and 5 days, respectively. Error bars represent the standard deviation from n = 3 independent experiments. (B and D) Immunoblotting analysis of CAL39 and SW954 cells showing increase in CHK1 phosphorylation after exposure to AZD7762 for 24 hours. β-actin is used as loading control. Representative data from n = 3 experiments

Article Snippet: Vulvar squamous cell carcinoma cell line CAL39 (DSMZ, Braunschweig Germany) was cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, LifeTechnologies, Invitrogen, Oslo, Norway) supplemented with 10% Fetal Calf Serum (Biocrom, KG, Berlin, Germany) and 2 mM Lglutamine (Lonza, Vervieres, Belgium) at 37°C in humidified condition containing 5% CO2.

Techniques: CTG Assay, Standard Deviation, Western Blot, Phospho-proteomics, Control

Journal: Cancer medicine

Article Title: Evaluation of CHK1 activation in vulvar squamous cell carcinoma and its potential as a therapeutic target in vitro.

doi: 10.1002/cam4.1638

Figure Lengend Snippet: FIGURE 6 CHK1 downregulation by siRNA leads to strong reduction in viability and apoptosis. (A) Effect of siCHK1 on cell viability of CAL39 and SW954 was assessed by trypan blue cell counting 3 days and 5 days after siRNA transfection, respectively. Error bars represent the standard deviation from n = 3 independent experiments. (B) Protein levels of downstream effector proteins following siRNA transfection by immunoblotting. β-actin was used as loading control. Representative data from n = 3 experiments

Article Snippet: Vulvar squamous cell carcinoma cell line CAL39 (DSMZ, Braunschweig Germany) was cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, LifeTechnologies, Invitrogen, Oslo, Norway) supplemented with 10% Fetal Calf Serum (Biocrom, KG, Berlin, Germany) and 2 mM Lglutamine (Lonza, Vervieres, Belgium) at 37°C in humidified condition containing 5% CO2.

Techniques: Cell Counting, Transfection, Standard Deviation, Western Blot, Control

Journal: Cancer medicine

Article Title: Evaluation of CHK1 activation in vulvar squamous cell carcinoma and its potential as a therapeutic target in vitro.

doi: 10.1002/cam4.1638

Figure Lengend Snippet: FIGURE 5 Effects of AZD7762 on protein expression of cell cycle regulators and apoptosis markers. Immunoblotting analysis of CAL39 and SW954 cells showing altered expression of proteins involved in cell cycle regulation and apoptosis after exposure to AZD7762 (500 nmol/L) for 24, 48, and 72 hours, respectively. β-actin is used as loading control. Representative data from n = 3 experiments

Article Snippet: Vulvar squamous cell carcinoma cell line CAL39 (DSMZ, Braunschweig Germany) was cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, LifeTechnologies, Invitrogen, Oslo, Norway) supplemented with 10% Fetal Calf Serum (Biocrom, KG, Berlin, Germany) and 2 mM Lglutamine (Lonza, Vervieres, Belgium) at 37°C in humidified condition containing 5% CO2.

Techniques: Expressing, Western Blot, Control

Journal: Gynecologic oncology

Article Title: Somatic mutation profiling of vulvar cancer: Exploring therapeutic targets.

doi: 10.1016/j.ygyno.2018.06.026

Figure Lengend Snippet: Fig. 1. Distribution of pathogenic mutations in hrHPV-positive (n = 52) and hrHPV-negative (n = 29) VSCC tumors. Each column corresponds to an individual tumor, while each row corresponds to the mutated gene. Tumor samples with no mutations detected are depicted in grey, missense mutations are highlighted in green, whereas black hits indicate nonsense mutations. The OncoPrinter tool from cBioPortal was used to generate this figure (http://www.cbioportal.org/oncoprinter.jsp). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The human vulvar squamous cell carcinoma (VSCC) cell line A431 (DSMZ, Germany) was cultured in DMEM (Dulbecco's modified Eagle's medium, Thermo Fisher Scientific) supplementedwith 10% fetal bovine serum (FBS, Thermo Fisher Scientific), and 1% penicillin/streptomycin Please cite this article as: S. Zięba, et al., Somaticmutation profiling of vulva doi.org/10.1016/j.ygyno.2018.06.026 antibiotics (AB, Thermo Fisher Scientific).

Techniques:

Journal: Gynecologic oncology

Article Title: Somatic mutation profiling of vulvar cancer: Exploring therapeutic targets.

doi: 10.1016/j.ygyno.2018.06.026

Figure Lengend Snippet: Fig. 2. Expression of total mTOR protein detected by IHC. A – HSIL sample, B – VSCC sample obtained from a patient with no progression, C - VSCC sample obtained from a patient experiencing disease progression, D – a sample of VSCC recurrence. Upper panel – 40× and lower panel – 10× magnification.

Article Snippet: The human vulvar squamous cell carcinoma (VSCC) cell line A431 (DSMZ, Germany) was cultured in DMEM (Dulbecco's modified Eagle's medium, Thermo Fisher Scientific) supplementedwith 10% fetal bovine serum (FBS, Thermo Fisher Scientific), and 1% penicillin/streptomycin Please cite this article as: S. Zięba, et al., Somaticmutation profiling of vulva doi.org/10.1016/j.ygyno.2018.06.026 antibiotics (AB, Thermo Fisher Scientific).

Techniques: Expressing, Biomarker Discovery

Journal: Gynecologic oncology

Article Title: Somatic mutation profiling of vulvar cancer: Exploring therapeutic targets.

doi: 10.1016/j.ygyno.2018.06.026

Figure Lengend Snippet: Fig. 3. Percentage of mTOR-staining tumor cells (upper panel) and immune infiltrating cells (lower panel) in HSIL and VSCC samples. Asterisks indicate the significance as follow: **** for b0.0001, *** for 0.0001 to 0.001, ** for 0.001 to 0.01 and * for 0.01 to 0.05 p-values.

Article Snippet: The human vulvar squamous cell carcinoma (VSCC) cell line A431 (DSMZ, Germany) was cultured in DMEM (Dulbecco's modified Eagle's medium, Thermo Fisher Scientific) supplementedwith 10% fetal bovine serum (FBS, Thermo Fisher Scientific), and 1% penicillin/streptomycin Please cite this article as: S. Zięba, et al., Somaticmutation profiling of vulva doi.org/10.1016/j.ygyno.2018.06.026 antibiotics (AB, Thermo Fisher Scientific).

Techniques: Staining

Journal: Gynecologic oncology

Article Title: Somatic mutation profiling of vulvar cancer: Exploring therapeutic targets.

doi: 10.1016/j.ygyno.2018.06.026

Figure Lengend Snippet: Fig. 4. Effect of rapamycin (red), everolimus (blue) and AZD2014 (yellow) at 10 μM concentrations on viability of A431, CAL-39 and SW-954 cells. The results are expressed as a mean of three experiments performed in three technical replicates. Asterisks indicate the significance of difference of cells viability (%) compared to control cells (white) according to the following convention: **** for b0.0001, *** for 0.0001 to 0.001, ** for 0.001 to 0.01 and * for 0.01 to 0.05 p-values. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The human vulvar squamous cell carcinoma (VSCC) cell line A431 (DSMZ, Germany) was cultured in DMEM (Dulbecco's modified Eagle's medium, Thermo Fisher Scientific) supplementedwith 10% fetal bovine serum (FBS, Thermo Fisher Scientific), and 1% penicillin/streptomycin Please cite this article as: S. Zięba, et al., Somaticmutation profiling of vulva doi.org/10.1016/j.ygyno.2018.06.026 antibiotics (AB, Thermo Fisher Scientific).

Techniques: Control

Journal: Gynecologic oncology

Article Title: Somatic mutation profiling of vulvar cancer: Exploring therapeutic targets.

doi: 10.1016/j.ygyno.2018.06.026

Figure Lengend Snippet: Fig. 5. Signaling pathways implicated in VSCC carcinogenesis. PI3KCA/AKT/mTOR and MAPK cellular pathways may be activated by RTK and/or by the mutated downstream members of these signaling circuits. Contained within the cytoplasm, these signaling network members function either as oncogenic (blue) or tumor suppressor (green) proteins. Genes encoding some of these proteins are frequently mutated in VSCC tumors, resulting in mTORC1 activation. In addition to the RB and TP53 mutations, two main cell cycle regulators may be inactivated by viral proteins produced in HPV infected individuals. Abbreviations: AKT - AKT Serine/Threonine Kinase 1; CDK4/6 - Cyclin Dependent Kinase 4/Cyclin Dependent Kinase 6; CDKN2A - Cyclin Dependent Kinase Inhibitor 2A; MAPK - Mitogen-Activated Protein Kinase; mTOR - Mechanistic Target Of Rapamycin Kinase; PI3KCA - Phosphatidylinositol-4,5- Bisphosphate 3-Kinase Catalytic Subunit Alpha; PTEN - Phosphatase And Tensin Homolog; FBWX7 - F-Box And WD Repeat Domain Containing 7; RAS – N-, K- and H-RAS; RB - RB Transcriptional Corepressor; RTK - receptor tyrosine kinases. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The human vulvar squamous cell carcinoma (VSCC) cell line A431 (DSMZ, Germany) was cultured in DMEM (Dulbecco's modified Eagle's medium, Thermo Fisher Scientific) supplementedwith 10% fetal bovine serum (FBS, Thermo Fisher Scientific), and 1% penicillin/streptomycin Please cite this article as: S. Zięba, et al., Somaticmutation profiling of vulva doi.org/10.1016/j.ygyno.2018.06.026 antibiotics (AB, Thermo Fisher Scientific).

Techniques: Protein-Protein interactions, Activation Assay, Produced, Infection