Journal: Cancer Medicine

Article Title: Evaluation of CHK 1 activation in vulvar squamous cell carcinoma and its potential as a therapeutic target in vitro

doi: 10.1002/cam4.1638

Figure Lengend Snippet: Expression of pCHK 1 Ser345 , pCHK 1 Ser317 , pCHK 1 Ser296 , and pCHK 1 Ser280 in vulvar squamous epithelium. Immunostaining of pCHK 1 Ser345 (A), pCHK 1 Ser317 (B), pCHK 1 Ser296 (C), and pCHK 1 Ser280 (D) in normal vulvar epithelium (Magnification × 300). High expression of pCHK 1 Ser345 (E), pCHK 1 Ser317 (F), pCHK 1 Ser296 (G), and pCHK 1 Ser280 (H) and low expression of pCHK 1 Ser345 (I), pCHK 1 Ser317 (J), pCHK 1 Ser296 (K), and pCHK 1 Ser280 (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 L‐glutamine (Lonza, Vervieres, Belgium) at 37°C in humidified condition containing 5% CO 2 .

Techniques: Expressing, Immunostaining

Journal: Oncotarget

Article Title: MiR-223-5p works as an oncomiR in vulvar carcinoma by TP63 suppression

doi: 10.18632/oncotarget.10247

Figure Lengend Snippet: ( A ) Relative expression of miR-223-5p was significantly increased when mimetics were added, compared to non-transfected – basal cells, detected by qRT-PCR. ( B ) MiR-223-5p expression remains significantly higher on transfected cells compared to basal SW962 for at least 96 hours post transfection. ** on behalf of p ≤ 0.01, and *** p ≤ 0.001.

Article Snippet: SW962 vulvar cancer cell line (derived from a vulvar carcinoma lymph node metastasis, HTB-118) was obtained from the American Type Culture Collection - ATCC (Manassas, VA, USA) and maintained routinely with RPMI 1640 media, (GIBCO ® , Life Tecnologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; GIBCO ® , Life technologies, Carlsbad, CA, USA) and 100 mg/mL Garamycin.

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: Oncotarget

Article Title: MiR-223-5p works as an oncomiR in vulvar carcinoma by TP63 suppression

doi: 10.18632/oncotarget.10247

Figure Lengend Snippet: Proliferation rates of SW962 cells were significantly reduced after mimetics microRNA were transfected compared to basal non-transfected cells at 72 hours and 96 hours post-transfection. ** on behalf of p ≤ 0.01 and * on behalf of p ≤ 0.05.

Article Snippet: SW962 vulvar cancer cell line (derived from a vulvar carcinoma lymph node metastasis, HTB-118) was obtained from the American Type Culture Collection - ATCC (Manassas, VA, USA) and maintained routinely with RPMI 1640 media, (GIBCO ® , Life Tecnologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; GIBCO ® , Life technologies, Carlsbad, CA, USA) and 100 mg/mL Garamycin.

Techniques: Transfection

Journal: Oncotarget

Article Title: MiR-223-5p works as an oncomiR in vulvar carcinoma by TP63 suppression

doi: 10.18632/oncotarget.10247

Figure Lengend Snippet: ( A ) Cells transfected with mimetic microRNA presented lower migration (higher wound area) compared to basal SW962 cells at 48 and 72 hours post-transfection. ( B ) Illustrative image of wound-healing assay at five time-points. ( C ) Higher number of invasive cells was observed when cells were transfected with miR-223-5p. *** on behalf of p ≤ 0.001 and * on behalf of p ≤ 0.05, n.s. on behalf of non-significant values.

Article Snippet: SW962 vulvar cancer cell line (derived from a vulvar carcinoma lymph node metastasis, HTB-118) was obtained from the American Type Culture Collection - ATCC (Manassas, VA, USA) and maintained routinely with RPMI 1640 media, (GIBCO ® , Life Tecnologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; GIBCO ® , Life technologies, Carlsbad, CA, USA) and 100 mg/mL Garamycin.

Techniques: Transfection, Migration, Wound Healing Assay

Journal: Oncotarget

Article Title: MiR-223-5p works as an oncomiR in vulvar carcinoma by TP63 suppression

doi: 10.18632/oncotarget.10247

Figure Lengend Snippet: ( A ) TP63 mRNA (bottom) contains a binding site from positions 956-977 for hsa-miR-233-5p (top). Mature microRNA is aligned with TP63 mRNA and hsa-miR-223-5p seed sequence is highlighted in red. ( B ) A significant reduction of p63 protein levels was observed when mimetics were added to SW962 cell line. Quantitative analysis and ( C ) illustrative image, being α-Tubulin was used as internal control. ( D ) Relative expression of TP63 by qRT-PCR normalized to HPRT was significantly lower on mimetics transfected cells compared to basal SW962 cells. *** on behalf of p ≤ 0.0001.

Article Snippet: SW962 vulvar cancer cell line (derived from a vulvar carcinoma lymph node metastasis, HTB-118) was obtained from the American Type Culture Collection - ATCC (Manassas, VA, USA) and maintained routinely with RPMI 1640 media, (GIBCO ® , Life Tecnologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; GIBCO ® , Life technologies, Carlsbad, CA, USA) and 100 mg/mL Garamycin.

Techniques: Binding Assay, Sequencing, Control, Expressing, Quantitative RT-PCR, Transfection

Journal: Oncotarget

Article Title: MiR-223-5p works as an oncomiR in vulvar carcinoma by TP63 suppression

doi: 10.18632/oncotarget.10247

Figure Lengend Snippet: ( A ) P63 is located in the nuclei of SW962 cells by immunofluorescence assay. From left to right, nuclei of vulvar SW962 cells are stained with DRAQ5 (Red); anti-p63 antibody is labeling cells for p63 protein (Green); merged image demonstrate nuclear expression of p63 (yellow). Magnification: 400×. ( B ) Representative images of p63 immunostaining (brown) in FFPE vulvar tumors, positive case (HScore < 150) on the left side and negative case (HScore ≥ 150) on the right side. Magnification: 200×. ( C ) Scatter dot plot demonstrate the distribution of patients with p63 positive and negative tumors versus miR-223-5p expression levels. Patients with negative p63 (left) presented statistically significant higher expression of miR-223-5p compared to the patients with p63 positivity * on behalf of p < 0.05.

Article Snippet: SW962 vulvar cancer cell line (derived from a vulvar carcinoma lymph node metastasis, HTB-118) was obtained from the American Type Culture Collection - ATCC (Manassas, VA, USA) and maintained routinely with RPMI 1640 media, (GIBCO ® , Life Tecnologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; GIBCO ® , Life technologies, Carlsbad, CA, USA) and 100 mg/mL Garamycin.

Techniques: Immunofluorescence, Staining, Labeling, Expressing, Immunostaining