Journal: Oncotarget

Article Title: Targeting growth hormone receptor in human melanoma cells attenuates tumor progression and epithelial mesenchymal transition via suppression of multiple oncogenic pathways

doi: 10.18632/oncotarget.15375

Figure Lengend Snippet: a . Representative images of western blot (WB) analyses of phosphorylation levels b . JAK2, c . STAT5, d . STAT1, e . STAT3, f . SRC, g . AKT, h . mTOR and i . ERK1/2, in excess human-GH treated or GHRKD human melanoma cell lysates. SK-MEL-28 cells, 24 hr post-transfection with either scramble (scr)-siRNA or GHR-siRNA were treated for ten mins with GH and lysed as described. WB was performed using appropriate antibodies. Densitometry analyses of individual blots was performed using ImageJ software and the ratio of phosphorylated vs. total protein levels against untreated scr-siRNA transfected controls. Overall, excess GH increased while GHRKD decreased phosphorylation states. Similar results for MALME-3M, MDA-MB-435 and SK-MEL-5 human melanoma cells are presented in . Blots from individual experiments were quantified and the mean of three blots per antibody was taken. Protein levels were normalized against expression of β-actin. [*, p < 0.05, Students t test, n = 3].

Article Snippet: Human malignant melanoma cell lines (part of NCI-60 panel of human cancer cells) - SK-MEL-5, SK-MEL-28, MALME-3M, MDA-MB-435S and normal human skin fibroblast cells MALME-3 cells were obtained from American Type Culture Collection (ATCC; Manassas, VA).

Techniques: Western Blot, Phospho-proteomics, Transfection, Software, Expressing

Journal: Oncotarget

Article Title: Targeting growth hormone receptor in human melanoma cells attenuates tumor progression and epithelial mesenchymal transition via suppression of multiple oncogenic pathways

doi: 10.18632/oncotarget.15375

Figure Lengend Snippet: a-c . Relative RNA expression was quantified for N-cadherin (a), vimentin (b) and E-cadherin (c) in SK-MEL-28 melanoma cells following addition of 5, 50 and 150 ng/mL hGH or 24 hr following GHRKD, in presence or absence of 0 and 50 ng/mL hGH. Similar results for MALME-3M, MDA-MB-435 and SK-MEL-5 human melanoma cells are presented in . In all cases, RNA expressions were normalized against β-actin and GAPDH values as reference genes and compared against untreated control. [*, p < 0.05, Wilcoxon sign rank test, n = 4] d-f . Densitometry analyses of relative protein expressions of N-cadherin (d), vimentin (e), and E-cadherin (f) as estimated by western blot (WB) of lysates of SK-MEL-28, MALME-3M, MDA-MB-435 and SK-MEL-5 cells, collected 60 hr post-transfection with either scramble (scr)-siRNA or GHR-siRNA in presence of GH. g . Representative images of WB analyses of phosphorylation levels of N-cadherin, vimentin and E-cadherin in four melanoma cell lines. WB was performed using appropriate antibodies. Densitometry analyses of individual blots was performed using ImageJ software and the ratio of phosphorylated vs. total protein levels against untreated scr-siRNA transfected controls. Overall, excess GH promoted while GHRKD reversed EMT in human melanoma cells. Blots from individual experiments were quantified and the mean of three blots per antibody was taken. Protein levels were normalized against expression of β-actin. [*, p < 0.05, Students t test, n = 3].

Article Snippet: Human malignant melanoma cell lines (part of NCI-60 panel of human cancer cells) - SK-MEL-5, SK-MEL-28, MALME-3M, MDA-MB-435S and normal human skin fibroblast cells MALME-3 cells were obtained from American Type Culture Collection (ATCC; Manassas, VA).

Techniques: RNA Expression, Control, Western Blot, Transfection, Phospho-proteomics, Software, Expressing

Journal: Oncotarget

Article Title: Targeting growth hormone receptor in human melanoma cells attenuates tumor progression and epithelial mesenchymal transition via suppression of multiple oncogenic pathways

doi: 10.18632/oncotarget.15375

Figure Lengend Snippet: a1 . Relative levels of RNA expressions for GH, GHR, PRL, PRLR, IGF1, IGFBP2, IGFBP3, IGF1R, IGF2R, IR, MET, ERBB3, and HGF expressed as 1/1000 th part of β-actin expression level in SK-MEL-28 melanoma cells. RNA levels of GH a2 . GHR a3 . PRL a4 . and PRLR a5 . as well as IGF1 b1 . IGFBP2 b2 . IGFBP3 b3 . IGF1R b4 . IGF2R b5 . and IR b6 . following RT-qPCR of RNA extracted from SK-MEL-28 cells following addition of 0, 5, 50 and 150 ng/mL hGH or following GHRKD, in presence or absence of 0 and 50 ng/mL hGH. Results are discussed in the text. Similar results for MALME-3M, MDA-MB-435 and SK-MEL-5 human melanoma cells are presented in , , , and . In all cases, exogenous hGH treatment was for 24 hr. RNA levels were normalized against expression of β-actin and GAPDH as reference genes. [*, p < 0.05, Wilcoxon sign rank test, n = 4].

Article Snippet: Human malignant melanoma cell lines (part of NCI-60 panel of human cancer cells) - SK-MEL-5, SK-MEL-28, MALME-3M, MDA-MB-435S and normal human skin fibroblast cells MALME-3 cells were obtained from American Type Culture Collection (ATCC; Manassas, VA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Oncotarget

Article Title: Targeting growth hormone receptor in human melanoma cells attenuates tumor progression and epithelial mesenchymal transition via suppression of multiple oncogenic pathways

doi: 10.18632/oncotarget.15375

Figure Lengend Snippet: Endocrine/paracrine/autocrine GH binds to GHR expressed at high levels in human malignant melanoma cells and activates JAK2 and SRC kinases. This leads to downstream activation of STAT1, STAT3, STAT5, ERK1/2, AKT and mTOR. Subsequent transcription of target genes lead to aggressive phenotypes of tumor cell migration, invasion and proliferation and upregulate autocrine HGF-MET loop, ERBB3 and also drives epithelial-mesenchymal-transition. In our study excess GH upregulated (in green) while siRNA mediated GHR knock-down (GHRKD) downregulated (in red) these effects.

Article Snippet: Human malignant melanoma cell lines (part of NCI-60 panel of human cancer cells) - SK-MEL-5, SK-MEL-28, MALME-3M, MDA-MB-435S and normal human skin fibroblast cells MALME-3 cells were obtained from American Type Culture Collection (ATCC; Manassas, VA).

Techniques: Activation Assay, Migration, Knockdown

Journal: BMC Cancer

Article Title: uPAR enhances malignant potential of triple-negative breast cancer by directly interacting with uPA and IGF1R

doi: 10.1186/s12885-016-2663-9

Figure Lengend Snippet: Co-overexpression of uPAS components and tumour-promoting proteins in TNBC samples and cell lines. a Immunohistochemical analysis of tumour samples showing positive expression and localisation of the proteins of interest: uPAR, uPA, PAI-1, IGF1R, IR and c-Met, bar: 50 μm and b Protein expressions of uPAR, uPA, PAI-1, IGF1R, IR and c-Met in the TNBC cohort ( n = 174). c Immunoblottings of uPAR, uPA (supernatant), PAI-1, IGF1R, (phospho) c-Met, HER2, ER, PR in two TNBC cell lines: BT549 and MDA-MB-231 and in the breast cancer cell lines: BT474, MCF7, MDA-MB-361, SKBR3 and T47D. Tubulin was used as loading control

Article Snippet: The following human breast cancer cells lines MDA-MB-361 (HTB-27), SKBR3 (HTB-30), T47D (HTB-133) were acquired from American Type Culture Collection (ATCC) and MCF7 (ACC115) cells from German Collection of Microorganisms and Cell Cultures, DSMZ).

Techniques: Over Expression, Immunohistochemical staining, Expressing, Control