Different prevalence of BRAF and NRAS somatic mutations in melanomas according to the patients’ origin.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e20013-e20013
Author(s):  
Giuseppe Palmieri ◽  
Amelia Lissia ◽  
Antonio Cossu ◽  
Paolo Antonio Ascierto ◽  
Gerardo Botti ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Somatic Mutations ◽  
Primary Melanoma ◽  
Cancer Genes ◽  
Nras Mutation ◽  
Braf Mutations ◽  
Paired Samples ◽  
Melanoma Metastases ◽  
Melanoma Cell Lines

e20013 Background: Genetic factors predisposing to melanoma at germline level have been demonstrated to be geographically heterogeneous. We here evaluated the spectrum of NRAS and BRAF mutations at somatic level, in a large subset of melanoma tissues from patients originating from different Italian geographical areas: Sardinia, whose population is genetically homogeneous, and Middle-South Italy, with a genetically heterogeneous population. Methods: Patients were enrolled consecutively between June 2008 and December 2012. Genomic DNA was isolated from tumor tissues [primary melanomas (N=439) and melanoma metastases (N=269)] or melanoma cell lines (N=32). Paired samples of primary melanomas (n=140) and synchronous or asynchronous metastases from the same patients (n=203) were included. The full coding sequences and splice junctions of NRAS (exons 2-3) and BRAF (exon 15) genes were screened for mutations through automated sequencing. Results: BRAF/NRAS mutations were identified in 63% of primary melanomas (48% BRAF; 15% NRAS), 67% melanoma metastases (51% BRAF; 16% NRAS), and 68% melanoma cell lines (56% BRAF; 12% NRAS). A non-significant increase in mutation frequency after progression from primary melanoma was observed. However, distribution of BRAF/NRAS mutations varied between in vivo tumors and melanoma cell lines, suggesting a preponderant role for BRAF activation in highly proliferating cultured melanoma cells. Among paired samples, consistency of BRAF/NRAS mutation patterns between metastatic and primary melanomas ranged from 71% (skin metastases) to about 90% (lymph node and visceral metastases). A significant inverse distribution of BRAF/NRAS mutation rates was observed in our series on the basis of the geographical origin of patients: for BRAF, 58% Sardinian vs. 42% non-Sardinian cases (p=0.045); for NRAS, 2% Sardinian vs. 21% non-Sardinian cases (p<0.001). Conclusions: Our findings provide additional insights into the spectrum and distribution of BRAF/NRAS mutations in melanoma; moreover, they support the hypothesis that differences in patients’ origins and related genetic backgrounds may contribute to even determine the incidence rate of somatic mutations in such cancer genes.

Braf Mutations Are Associated With High Levels of Phosphorylated RKIP in Melanoma Cell Lines: Potential Prognostic Significance

2011 ◽  
Vol 2 (2) ◽  
pp. 189-194 ◽  
Author(s):  
Sara Huerta-Yepez ◽  
S. Ekmekcioglu ◽  
C. M. Rivera-Pazos ◽  
G. Antonio-Andres ◽  
Mario I. Vega ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Prognostic Significance ◽  
Braf Mutations ◽  
Melanoma Cell Lines

scholarly journals The engagement of CTLA-4 on primary melanoma cell lines induces antibody-dependent cellular cytotoxicity and TNF-α production

2013 ◽  
Vol 11 (1) ◽  
pp. 108 ◽  
Author(s):  
Stefania Laurent ◽  
Paola Queirolo ◽  
Silvia Boero ◽  
Sandra Salvi ◽  
Patrizia Piccioli ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Primary Melanoma ◽  
Cellular Cytotoxicity ◽  
Antibody Dependent Cellular Cytotoxicity ◽  
Tnf Α ◽  
Melanoma Cell Lines

Androgen receptors in human melanoma cell lines IIB-MEL-LES and IIB-MEL-IAN and in human melanoma metastases

2002 ◽  
Vol 12 (6) ◽  
pp. 529-538 ◽  
Author(s):  
V Morvillo ◽  
I A Lüthy ◽  
A I Bravo ◽  
M I Capurro ◽  
P Portela ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Androgen Receptors ◽  
Human Melanoma ◽  
Human Melanoma Cell ◽  
Melanoma Metastases ◽  
Melanoma Cell Lines

scholarly journals Unbiased High-Throughput Drug Combination Pilot Screening Identifies Synergistic Drug Combinations Effective against Patient-Derived and Drug-Resistant Melanoma Cell Lines

2020 ◽  
pp. 247255522097091
Author(s):  
David A. Close ◽  
John M. Kirkwood ◽  
Ronald J. Fecek ◽  
Walter J. Storkus ◽  
Paul A. Johnston
Keyword(s):  
Growth Inhibition ◽  
Cell Lines ◽  
Melanoma Cell ◽  
High Throughput ◽  
Drug Combination ◽  
High Throughput Screening ◽  
Dna Methyltransferase ◽  
Cross Resistance ◽  
Braf Mutations ◽  
Melanoma Cell Lines

We describe the development, optimization, and validation of 384-well growth inhibition assays for six patient-derived melanoma cell lines (PDMCLs), three wild type (WT) for BRAF and three with V600E- BRAF mutations. We conducted a pilot drug combination (DC) high-throughput screening (HTS) of 45 pairwise 4×4 DC matrices prepared from 10 drugs in the PDMCL assays: two B-Raf inhibitors (BRAFi), a MEK inhibitor (MEKi), and a methylation agent approved for melanoma; cytotoxic topoisomerase II and DNA methyltransferase chemotherapies; and drugs targeting the base excision DNA repair enzyme APE1 (apurinic/apyrimidinic endonuclease-1/redox effector factor-1), SRC family tyrosine kinases, the heat shock protein 90 (HSP90) molecular chaperone, and histone deacetylases. Pairwise DCs between dasatinib and three drugs approved for melanoma therapy—dabrafenib, vemurafenib, or trametinib—were flagged as synergistic in PDMCLs. Exposure to fixed DC ratios of the SRC inhibitor dasatinib with the BRAFis or MEKis interacted synergistically to increase PDMCL sensitivity to growth inhibition and enhance cytotoxicity independently of PDMCL BRAF status. These DCs synergistically inhibited the growth of mouse melanoma cell lines that either were dabrafenib-sensitive or had acquired resistance to dabrafenib with cross resistance to vemurafenib, trametinib, and dasatinib. Dasatinib DCs with dabrafenib, vemurafenib, or trametinib activated apoptosis and increased cell death in melanoma cells independently of their BRAF status or their drug resistance phenotypes. These preclinical in vitro studies provide a data-driven rationale for the further investigation of DCs between dasatinib and BRAFis or MEKis as candidates for melanoma combination therapies with the potential to improve outcomes and/or prevent or delay the emergence of disease resistance.

Neurotrophin signaling and melanoma brain metastases.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e13024-e13024
Author(s):  
Nicole Fortenbery ◽  
Rajappa Kenchappa ◽  
Peter A. J. Forsyth
Keyword(s):  
Brain Metastases ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Neurotrophin Receptor ◽  
Neurotrophin Receptors ◽  
Melanoma Metastases ◽  
Melanoma Brain Metastases ◽  
Neurotrophin Signaling ◽  
The Brain ◽  
Melanoma Cell Lines

e13024 Background: Adult metastatic brain tumors occur more frequently than primary intracranial neoplasms. Melanoma is the third most common tumor type that metastasizes to the brain. Therefore, elucidating the underlying biological mechanisms of melanoma metastases is critical. Melanocytes and neurons share a neural ectodermal origin, and thus, melanoma may preferentially travel to the brain due to the expression of common neurotrophin receptors, namely p75 neurotrophin receptor (p75NTR) and Trks. Further, high concentrations of neurotrophic factors present in the brain may recruit metastatic melanoma. We hypothesize that neurotrophin signaling through p75NTR is required for the malignant phenotype of melanoma and melanoma brain metastases (MBM). Methods: We investigated the expression of several neutrophin receptors, signaling molecules, and neurotrophins that we hypothesize to be important in the process of MBM. Using 14 malignant melanoma cell lines and two primary MBMs grown under neurosphere conditions, we investigated the expression of these molecules using standard western blotting, flow cytometry, and RT-PCR. We also performed an in depth microarray using of 14 primary MBM patients from Moffitt’s Total Cancer Care project. Results: All melanoma lines examined have robust expression of p75NTR, Trks, and neurotrophins. We find melanoma cell lines resistant to the BRAF inhibitor, vemurafenib, have elevated expression of neurotrophins and neurotrophin receptors when compared to lines sensitive to vemurafenib. We also detect high levels of p75NTR in patient MBM, at both mRNA and protein levels. Finally, we find a significant level of expression of all 11 genes tested by microarray. Conclusions: Previous studies have demonstrated that p75NTR has a role in melanoma cell survival in vitro. Here we demonstrate p75NTR and Trk receptors have high expression in melanoma cell lines and importantly, primary MBM. These data suggest that signaling through neurotophin receptors may be important for melanoma metastases and survival in the brain. Moreover, increased expression by vemurafenib resistant lines may suggest that these cells upregulate the expression of neurotrophin signaling as a means of treatment resistant.

Metastatic but not primary melanoma cell lines growin vitro independently of exogenous growth factors

1987 ◽  
Vol 40 (5) ◽  
pp. 687-690 ◽  
Author(s):  
Ulrich Rodeck ◽  
Meenhard Herlyn ◽  
Hans D. Menssen ◽  
Richard W. Furlanetto ◽  
Hilary Koprowski
Keyword(s):  
Growth Factors ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Primary Melanoma ◽  
Exogenous Growth ◽  
Exogenous Growth Factors ◽  
Melanoma Cell Lines

Expression of Bcl-2 family members in human melanocytes, in melanoma metastases and in melanoma cell lines

1998 ◽  
Vol 8 (3) ◽  
pp. 197-203 ◽  
Author(s):  
E Selzer ◽  
H Schlagbauer-Wadl ◽  
I Okamoto ◽  
H Pehamberger ◽  
R P??tter ◽  
...  
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Family Members ◽  
Human Melanocytes ◽  
Melanoma Metastases ◽  
Melanoma Cell Lines

scholarly journals Comprehensive upstream and downstream regulatory analyses identify miR-675-3p as a potential prognostic biomarker in melanoma

Human Cell ◽  
2021 ◽  
Author(s):  
Cai-Chou Zhao ◽  
Hao Guo ◽  
Ying Wang ◽  
Jiu-Hong Li
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Peripheral Blood ◽  
Melanoma Patient ◽  
Target Genes ◽  
Prognostic Biomarker ◽  
Primary Melanoma ◽  
Melanoma Patients ◽  
Melanoma Cell Lines

AbstractThis study assessed miR-675-3p-related regulatory mechanisms in melanoma and the clinical relevance of such regulatory activities. We downloaded miRNA mature strand expression RNA-Seq, phenotypic, and DNA methylation data pertaining to the TCGA Melanoma cohort. Differentially expressed miRNAs (DEMs) between metastatic and primary melanoma patient tissues were then identified, and miR-675-3p expression in melanoma patient peripheral blood was confirmed using the GSE20994 GEO dataset, while its expression in melanoma cell lines was evaluated via qRT-RCR. The clinical and prognostic implications of miR-675-3p in melanoma were assessed, and miR-675-3p target genes were identified using bioinformatics tools. Functional roles of this miRNA were explored via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. We identified 3 and 22 miRNAs that were up- and downregulated, respectively, in metastatic melanoma samples relative to primary melanoma samples. Upregulation of miR-675-3p was associated with poorer overall patient survival, tumor histologic grade, and Clark's level. Consistently, miR-675-3p was also overexpressed in the peripheral blood of melanoma patients relative to healthy controls, and in melanoma cell lines relative to control cells. Gene regulatory networks indicated that 32 transcription factors control miR-675-3p expression, and that it, in turn, regulates 10 target genes. KEGG analyses indicated that these genes were associated with cell cycle, transcriptional misregulation in cancer, TGF-beta signaling, and HIF-1 signaling pathways. Gain-of-function assays revealed that miR-675-3p could promote cell proliferation via accelerating cell cycle progression. Western blotting results indicated that miR-675-3p could active TGF-beta and HIF-1 signaling. Through upstream and downstream analyses of miR-675-3p-related regulatory activity, we confirmed that this miRNA participates in key melanoma-related processes and offers value as a prognostic biomarker in melanoma patients.

Skp2, a prognostic marker and potential therapeutic target in metastatic melanoma

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. 11034-11034
Author(s):  
G. Wang ◽  
D. Hanniford ◽  
A. Rose ◽  
A. Gaziel ◽  
A. Pavlick ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Cell Lines ◽  
Melanoma Cell ◽  
Primary Melanoma ◽  
Melanoma Metastasis ◽  
Genomic Amplification ◽  
Aberrant Expression ◽  
Skp2 Protein ◽  
Melanoma Patients ◽  
Melanoma Cell Lines

11034 Background: Skp2, a known oncogene, is overexpressed in several types of tumors and is associated with worse recurrence rate and overall survival in primary melanoma patients. Moreover, the anti-proliferative effects of Skp2 siRNA on various tumor cell lines have prompted the preclinical testing of Skp2 small molecule inhibitors. In this study, we assessed the clinical relevance and molecular mechanism(s) underlying Skp2 overexpression in metastatic melanoma patients. Methods: Skp2 protein levels were measured in 122 metastatic melanoma specimens using immunohistochemistry (IHC), and the association between Skp2 overexpression and post-recurrence survival was examined. Moreover, 22 cell lines (2 normal primary melanocytes, 2 primary immortal melanocytes, 4 primary melanoma cell lines, and 18 metastatic melanoma cell lines) were evaluated for Skp2 genomic amplification using Single Nucleotide Polymorphism (SNP) arrays (Affymetrix 6.0) and Skp2 gene expression using mRNA arrays (Affymetrix U133A 2.0) and quantitative RT-PCR. We also screened 18 cell lines for Skp2 mutation by sequencing. Results: Skp2 overexpression, defined as >25% tumor cells, was associated with shorter 3-yr post-recurrence survival (37%) compared to Skp2 expression ≤25% (55%) (HR=1.89, 95%, CI= 1.04, 3.42, p=0.04). Skp2 overexpression was significantly associated with the site of melanoma metastasis: visceral (n= 12; 89%), lymph node (n=49; 36%), brain (n=15; 14%), and soft-tissue (n=36; 6%) (p<0.001). SNP array revealed genomic amplification at the Skp2 locus in 6 (33%) metastatic cell lines and one primary melanoma cell line. Skp2 genomic amplification was associated with increased transcript expression. No Skp2 mutations were identified. Conclusions: Skp2 protein overexpression is associated with worse prognosis in metastasis in melanoma. Our results also support that gene amplification, rather than a Skp2 gene mutation, may be the major mechanism responsible for Skp2 aberrant expression in metastatic melanoma. No significant financial relationships to disclose.

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