Multiplex ligation-dependent probe amplification of conjunctival melanoma reveals common BRAF V600E gene mutation and gene copy number changes

Invest Ophthalmol Vis Sci. 2011 Jul 29;52(8):5598-604. doi: 10.1167/iovs.10-6934.

Abstract

Purpose: To determine the occurrence of BRAF V600E gene mutations and copy number changes of all autosome arms and genes known to be frequently altered in tumorigenesis in primary and metastatic conjunctival melanomas (CoMs).

Methods: DNA (200 ng) was analyzed by three multiplex ligation-dependent probe amplification assays (P027 uveal melanoma, P036 human telomere, and P206 spitzoid melanoma).

Results: Eight of 16 primary tumor samples and 4 of 6 metastatic samples showed BRAF V600E gene mutations. CDKN1A and RUNX2 (both 6p21.2) were amplified in 11 and 16 of 21 primary CoMs, respectively. In metastatic CoMs, MLH1 (3p22.1) and TIMP2 (17q25.3) were frequently amplified, and MGMT (20q26.3) and ECHS1 (10q26.3) were frequently deleted. The BDH (3q), FLJ20265 (4p), OPRL1 (20q), and PAO (10q) genes, representing the telomeres of their respective chromosome arms in the P036 assay, were frequently amplified in metastatic CoMs. No statistically significant associations were identified between BRAF mutation or CDKN1A or RUNX2 amplification and sex, age, histologic cell type, or patient survival.

Conclusions: No copy number changes were associated exclusively with metastatic CoMs. However, further investigation of the role of CDKN1A and RUNX2 in CoMs development and that of MLH1, TIMP2, MGMT, and ECHS1 in metastatic CoMs is warranted. Validation of the observed gene and chromosome arm copy number changes in a larger cohort of primary and metastatic CoMs is necessary to identify the patients at highest risk for CoMs metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Conjunctival Neoplasms / epidemiology
  • Conjunctival Neoplasms / genetics*
  • Conjunctival Neoplasms / pathology
  • DNA Probes
  • Female
  • Gene Dosage / genetics*
  • Genetic Predisposition to Disease / epidemiology
  • Genetic Predisposition to Disease / genetics
  • Humans
  • Male
  • Melanoma / epidemiology
  • Melanoma / genetics*
  • Melanoma / secondary
  • Middle Aged
  • Nucleic Acid Amplification Techniques / methods
  • Point Mutation*
  • Prognosis
  • Proto-Oncogene Proteins B-raf / genetics*
  • Risk Factors
  • Young Adult

Substances

  • DNA Probes
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf