Detection of mutated BRAFV600E variant in circulating DNA of stage III-IV melanoma patients

Maria Daniotti, Viviana Vallacchi, Licia Rivoltini, Roberto Patuzzo, Mario Santinami, Flavio Arienti, Gianluca Cutolo, Marco A. Pierotti, Giorgio Parmiani, Monica Rodolfo

Research output: Contribution to journalArticle

Abstract

BRAFV600E is the most represented somatic point mutation in cutaneous melanoma, thus providing a unique molecular marker for this disease. The development of efficient methods for its detection in free circulating DNA of patients may lead to the improvement of diagnostic and prognostic tools. With this aim, we evaluated whether BRAFV600E represents a detectable marker in the plasma/serum from melanoma patients in a pilot study. Circulating cell-free DNA was extracted from the serum or plasma of 15 healthy donors and 41 melanoma patients at different clinical stages and obtained either presurgery or after surgery during follow-up. Quantitative analysis showed higher levels of circulating free DNA in patients compared to controls, with the highest levels detected in samples obtained presurgery and at stage IV. Four different PCR methods were compared for their capacity to amplify a few copies of BRAFV600E in wild-type DNA. BRAFV600E was detectable in circulating DNA of 12 patients and in none of the controls; only 1 PCR method reproducibly amplified BRAFV600E. Positive samples were obtained from 8/13 patients at stage IV and from 4/24 patients at stage III, but not in 4 patients at stage I-II; half of the positives were obtained presurgery and half at follow-up. Correspondence between circulating DNA and related tumors were examined for 20 patients, and a correlation was found for stage IV patients. In conclusion, this method can be utilized for monitoring the disease in stage IV melanoma patients but it appears unsatisfactory for the early detection of melanoma.

Original languageEnglish
Pages (from-to)2439-2444
Number of pages6
JournalInternational Journal of Cancer
Volume120
Issue number11
DOIs
Publication statusPublished - Jun 1 2007

Fingerprint

Melanoma
DNA
Polymerase Chain Reaction
Serum
Point Mutation
Tissue Donors
Skin

Keywords

  • BRAF
  • Circulating DNA
  • Melanoma
  • Plasma tumor marker

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Detection of mutated BRAFV600E variant in circulating DNA of stage III-IV melanoma patients. / Daniotti, Maria; Vallacchi, Viviana; Rivoltini, Licia; Patuzzo, Roberto; Santinami, Mario; Arienti, Flavio; Cutolo, Gianluca; Pierotti, Marco A.; Parmiani, Giorgio; Rodolfo, Monica.

In: International Journal of Cancer, Vol. 120, No. 11, 01.06.2007, p. 2439-2444.

Research output: Contribution to journalArticle

Daniotti, Maria ; Vallacchi, Viviana ; Rivoltini, Licia ; Patuzzo, Roberto ; Santinami, Mario ; Arienti, Flavio ; Cutolo, Gianluca ; Pierotti, Marco A. ; Parmiani, Giorgio ; Rodolfo, Monica. / Detection of mutated BRAFV600E variant in circulating DNA of stage III-IV melanoma patients. In: International Journal of Cancer. 2007 ; Vol. 120, No. 11. pp. 2439-2444.
@article{5e34fefcba8d4a939ec0ede00c1f5c75,
title = "Detection of mutated BRAFV600E variant in circulating DNA of stage III-IV melanoma patients",
abstract = "BRAFV600E is the most represented somatic point mutation in cutaneous melanoma, thus providing a unique molecular marker for this disease. The development of efficient methods for its detection in free circulating DNA of patients may lead to the improvement of diagnostic and prognostic tools. With this aim, we evaluated whether BRAFV600E represents a detectable marker in the plasma/serum from melanoma patients in a pilot study. Circulating cell-free DNA was extracted from the serum or plasma of 15 healthy donors and 41 melanoma patients at different clinical stages and obtained either presurgery or after surgery during follow-up. Quantitative analysis showed higher levels of circulating free DNA in patients compared to controls, with the highest levels detected in samples obtained presurgery and at stage IV. Four different PCR methods were compared for their capacity to amplify a few copies of BRAFV600E in wild-type DNA. BRAFV600E was detectable in circulating DNA of 12 patients and in none of the controls; only 1 PCR method reproducibly amplified BRAFV600E. Positive samples were obtained from 8/13 patients at stage IV and from 4/24 patients at stage III, but not in 4 patients at stage I-II; half of the positives were obtained presurgery and half at follow-up. Correspondence between circulating DNA and related tumors were examined for 20 patients, and a correlation was found for stage IV patients. In conclusion, this method can be utilized for monitoring the disease in stage IV melanoma patients but it appears unsatisfactory for the early detection of melanoma.",
keywords = "BRAF, Circulating DNA, Melanoma, Plasma tumor marker",
author = "Maria Daniotti and Viviana Vallacchi and Licia Rivoltini and Roberto Patuzzo and Mario Santinami and Flavio Arienti and Gianluca Cutolo and Pierotti, {Marco A.} and Giorgio Parmiani and Monica Rodolfo",
year = "2007",
month = "6",
day = "1",
doi = "10.1002/ijc.22598",
language = "English",
volume = "120",
pages = "2439--2444",
journal = "International Journal of Cancer",
issn = "0020-7136",
publisher = "Wiley-Liss Inc.",
number = "11",

}

TY - JOUR

T1 - Detection of mutated BRAFV600E variant in circulating DNA of stage III-IV melanoma patients

AU - Daniotti, Maria

AU - Vallacchi, Viviana

AU - Rivoltini, Licia

AU - Patuzzo, Roberto

AU - Santinami, Mario

AU - Arienti, Flavio

AU - Cutolo, Gianluca

AU - Pierotti, Marco A.

AU - Parmiani, Giorgio

AU - Rodolfo, Monica

PY - 2007/6/1

Y1 - 2007/6/1

N2 - BRAFV600E is the most represented somatic point mutation in cutaneous melanoma, thus providing a unique molecular marker for this disease. The development of efficient methods for its detection in free circulating DNA of patients may lead to the improvement of diagnostic and prognostic tools. With this aim, we evaluated whether BRAFV600E represents a detectable marker in the plasma/serum from melanoma patients in a pilot study. Circulating cell-free DNA was extracted from the serum or plasma of 15 healthy donors and 41 melanoma patients at different clinical stages and obtained either presurgery or after surgery during follow-up. Quantitative analysis showed higher levels of circulating free DNA in patients compared to controls, with the highest levels detected in samples obtained presurgery and at stage IV. Four different PCR methods were compared for their capacity to amplify a few copies of BRAFV600E in wild-type DNA. BRAFV600E was detectable in circulating DNA of 12 patients and in none of the controls; only 1 PCR method reproducibly amplified BRAFV600E. Positive samples were obtained from 8/13 patients at stage IV and from 4/24 patients at stage III, but not in 4 patients at stage I-II; half of the positives were obtained presurgery and half at follow-up. Correspondence between circulating DNA and related tumors were examined for 20 patients, and a correlation was found for stage IV patients. In conclusion, this method can be utilized for monitoring the disease in stage IV melanoma patients but it appears unsatisfactory for the early detection of melanoma.

AB - BRAFV600E is the most represented somatic point mutation in cutaneous melanoma, thus providing a unique molecular marker for this disease. The development of efficient methods for its detection in free circulating DNA of patients may lead to the improvement of diagnostic and prognostic tools. With this aim, we evaluated whether BRAFV600E represents a detectable marker in the plasma/serum from melanoma patients in a pilot study. Circulating cell-free DNA was extracted from the serum or plasma of 15 healthy donors and 41 melanoma patients at different clinical stages and obtained either presurgery or after surgery during follow-up. Quantitative analysis showed higher levels of circulating free DNA in patients compared to controls, with the highest levels detected in samples obtained presurgery and at stage IV. Four different PCR methods were compared for their capacity to amplify a few copies of BRAFV600E in wild-type DNA. BRAFV600E was detectable in circulating DNA of 12 patients and in none of the controls; only 1 PCR method reproducibly amplified BRAFV600E. Positive samples were obtained from 8/13 patients at stage IV and from 4/24 patients at stage III, but not in 4 patients at stage I-II; half of the positives were obtained presurgery and half at follow-up. Correspondence between circulating DNA and related tumors were examined for 20 patients, and a correlation was found for stage IV patients. In conclusion, this method can be utilized for monitoring the disease in stage IV melanoma patients but it appears unsatisfactory for the early detection of melanoma.

KW - BRAF

KW - Circulating DNA

KW - Melanoma

KW - Plasma tumor marker

UR - http://www.scopus.com/inward/record.url?scp=34247337993&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34247337993&partnerID=8YFLogxK

U2 - 10.1002/ijc.22598

DO - 10.1002/ijc.22598

M3 - Article

C2 - 17315191

AN - SCOPUS:34247337993

VL - 120

SP - 2439

EP - 2444

JO - International Journal of Cancer

JF - International Journal of Cancer

SN - 0020-7136

IS - 11

ER -