Incidence of <b>
                     <i>BRAF</i>
                  </b> Oncogene Mutation and Clinical Relevance for Primary Cutaneous Melanomas

Shinozaki, Masaru; Fujimoto, Akihide; Morton, Donald L.; Hoon, Dave S.B.

doi:10.1158/1078-0432.ccr-1169-3

Cited by 208 publications

(211 citation statements)

References 15 publications

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“…Furthermore, it has been previously shown using the same technique that the frequency of BRAFV600E mutations in primary cutaneous melanoma can vary greatly with incidences as low as 30% being reported. 11 Our own previous experience has shown that the frequency could be even lower. 8 In spite of the low incidence rate, findings from this preliminary study shed light on the potential use of immunohistochemistry with the anti-B-Raf (V600E) antibody as an ancillary screening tool to assess the BRAFV600E mutation status in primary cutaneous melanoma.…”

Section: Discussionmentioning

confidence: 96%

“…In early melanomas, the BRAF mutation status has not been shown to affect overall survival; however, in metastatic melanoma, it has been associated with a poorer survival rate. 11,12 With regard to papillary thyroid carcinomas, evidence suggests that molecular testing for BRAFV600E in thyroid fineneedle aspirations can be a useful tool to separate this malignancy from other thyroid nodules. 13 The mutation status also serves as a prognostic marker as its presence is associated with higher rates of tumor recurrence and mortality.…”

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confidence: 99%

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Immunohistochemistry with a mutation-specific monoclonal antibody as a screening tool for the BRAFV600E mutational status in primary cutaneous malignant melanoma

2013

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The V600E mutation of BRAF has emerged as both an effective biomarker and therapeutic target for select benign and malignant cutaneous and non-cutaneous human tumors and is typically determined using DNAbased techniques that include allele-specific PCR and direct DNA sequencing. Recently however, the development of new antibodies directed against the V600E protein has opened the door for an easier and more efficient strategy for identifying this mutation. Our present aim was to determine the efficacy of one such antibody, anti-B-Raf (V600E), a mouse monoclonal antibody in which the immunogen is a synthetic peptide derived from the internal region of BRAFV600E. A total of 35 cases of primary cutaneous melanoma were evaluated using a combination of DNA-based techniques that included allele-specific PCR and/or direct DNA sequencing and immunohistochemistry. Cases of papillary thyroid carcinomas (n ¼ 5) and colorectal carcinomas (n ¼ 5), known to harbor the BRAFV600E mutation, served as positive controls for the study. DNA analyses revealed that 6 of 35 (17%) cases of the primary cutaneous malignant melanoma possessed the BRAFV600E mutation. For immunohistochemical analyses, cytoplasmic positivity with anti-B-Raf was noted in 7 of 35 (20%) cases of primary melanoma and in all 10 positive controls. Statistical analyses of the data demonstrated that the sensitivity of the immunohistochemistry was 100% and specificity was 97%. Findings from the current study support the potential use of immunohistochemistry as an ancillary screening tool to assess the BRAFV600E mutation status in primary cutaneous melanoma.

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Section: Discussionmentioning

confidence: 96%

mentioning

confidence: 99%

Immunohistochemistry with a mutation-specific monoclonal antibody as a screening tool for the BRAFV600E mutational status in primary cutaneous malignant melanoma

2013

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“…These findings are in keeping with other reports, 15 and further corroborate the notion that histological subtype is not a surrogate for BRAF mutation status and that it is less informative than other parameters, such as patient age, anatomical site of the primary neoplasm, and degree of solar elastosis at the primary site. 19,20 A number of molecular approaches are widely used in clinical practice to detect BRAF mutations in melanoma. These methods include (but are not restricted to) melting-curve analysis of PCR products, PCR-based DNA sequencing such as pyrosequencing and Sanger sequencing, 2,15,16,21e23 , as well as high-throughput methods based on DNA primer extension, such as MALDI-TOF MS (a SNP array) and next-generation sequencing platforms.…”

Section: Discussionmentioning

confidence: 99%

Frequency and Spectrum of BRAF Mutations in a Retrospective, Single-Institution Study of 1112 Cases of Melanoma

Greaves

Verma

Patel

et al. 2013

The Journal of Molecular Diagnostics

201

139

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The US Food and Drug Administration (FDA) approved vemurafenib to treat patients with metastatic melanoma harboring the BRAF c.1799T>A (p.V600E) mutation. However, a subset of melanomas harbor non-p.V600E BRAF mutations, and these data are of potential importance regarding the efficacy of current targeted therapies. To better understand the BRAF mutation profile in melanomas, we retrospectively analyzed data from 1112 primary and metastatic melanomas at our institution. The cohort included nonacral cutaneous (n Z 774), acral (n Z 111), mucosal (n Z 26), uveal (n Z 23), leptomeningeal (n Z 1), and metastatic melanomas of unknown primary site (n Z 177). BRAF mutation hotspot regions in exons 11 and 15 were analyzed by pyrosequencing or with the primer extension MassARRAY system. A total of 499 (44.9%) specimens exhibited BRAF mutations, involving exon 15 [497 (99.6%)] or exon 11 [2 (0.4%)]. p.V600E was detected in 376 (75.4%) cases; the remaining 123 (24.6%) cases exhibited non-p.V600E mutations, of which p.V600K was most frequent [86 (17.2%)]. BRAF mutations were more frequent in nonacral cutaneous (51.4%) than acral melanomas [18 (16.2%)] (P < 0.001); however, there was no significant difference among cutaneous histological subtypes. All mucosal, uveal, and leptomeningeal melanomas were BRAF wild type (WT). The high frequency of non-p.V600E BRAF mutations in melanoma has important implications because the FDA-approved companion diagnostic test for p.V600E detects some but not all non-p.V600E mutations. However, the therapeutic efficacy of vemurafenib is not well established in these lesions. (J Mol Diagn 2013, 15: 220e226; http://dx

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“…An activating mutation of v-raf murine sarcoma viral oncogene homolog B1 (BRaf) (mutation: V600E, also less commonly: V600K) 52 , which encodes for a protein kinase that regulates cell growth, is present in 50-70% of melanomas 53 . It is thought that mutant BRaf becomes locked into an activated mode, sending a continuous proliferative signal to the cell through the mitogen activated protein kinase (MAPK) pathway, which also serves to lower the levels of the pro-apoptotic protein Bcl-2 interacting mediator (Bim) which is targeted for degradation by the MAPK pathway 54,55 .…”

Section: Chemoresistance and Treatment In Malignant Melanomamentioning

confidence: 99%

Role of the pro-survival molecule Bfl-1 in melanoma

Hind

Carter

Harris

et al. 2015

The International Journal of Biochemistry & Cell Biology

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Melanoma, the cancer derived from the melanocytes of the skin, is becoming an ever more common cancer in the ageing population of the developed world and displays an intrinsic resistance to current therapies. This chemo resistance makes metastatic melanoma an extremely difficult cancer to treat leading to a 5 year survival rate of just 20%. As such, it is important to unearth new potential drug targets to increase the prospects for melanoma patients.Bfl-1 is a pro-survival protein of the Bcl-2 family of apoptosis regulating proteins. It has been found to be over-expressed in a small subset of chemo resistant tumours, including melanoma. Accordingly, I characterised the molecule in melanoma cells and determined its role in protecting these cells from chemotherapy-induced apoptosis. I elucidated the expression profile and half-lives of the protein and mRNA across a panel of melanoma cell-lines and healthy melanocytes. I also confirmed, using pathway specific inhibitors, that Bfl-1 was transcriptionally regulated by the NFB pathway and concluded through pulsechase experiments that Bfl-1 protein was degraded, at least in part, by the proteasome. Subcellular fractionation and indirect immunofluorescence techniques elucidated that Bfl-1 was located mostly at the mitochondria in both resting and apoptotic cells, with a diffuse level present throughout the cytoplasm. As well as characterising the protein in both melanoma cells and healthy primary melanocytes, I determined its role in the resistance of these cells to apoptosis. Over-expressed Bfl-1 was found to protect melanoma cells from apoptosis caused by a range of chemotherapeutic agents in A375 melanoma cells, which naturally expressed very low levels of Bfl-1. Further, knock down of Bfl-1 using siRNA technology in melanoma cells revealed the dependence of these cells on Bfl-1 for their resistance to certain chemotherapeutic agents currently used in melanoma treatment, including the MEK inhibitor PD901. All together, this research contributes to our understanding of Bfl-1 and highlights it as a potentially important therapeutic target in metastatic melanoma.

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Incidence of BRAF Oncogene Mutation and Clinical Relevance for Primary Cutaneous Melanomas

Cited by 208 publications

References 15 publications

Immunohistochemistry with a mutation-specific monoclonal antibody as a screening tool for the BRAFV600E mutational status in primary cutaneous malignant melanoma

Immunohistochemistry with a mutation-specific monoclonal antibody as a screening tool for the BRAFV600E mutational status in primary cutaneous malignant melanoma

Frequency and Spectrum of BRAF Mutations in a Retrospective, Single-Institution Study of 1112 Cases of Melanoma

Role of the pro-survival molecule Bfl-1 in melanoma

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