Quantitative in situ evaluation of telomeres in fluorescence in situ hybridization-processed sections of cutaneous melanocytic lesions and correlation with telomerase activity.
BACKGROUND
Telomere length is correlated with cellular ageing and immortalization processes. In some human cancers telomere length measurement has proved to be of diagnostic and prognostic value. Results comparable with the traditional terminal restriction fragment length determination by Southern blotting have been obtained in metaphase and interphase cells in some studies by fluorescence in situ hybridization (FISH) analysis; FISH additionally allows for the quantification of telomeres at the cellular level.


OBJECTIVE
In this study, 32 melanocytic lesions were analysed by FISH, aiming at investigating possible telomere differences among various benign and malignant lesions and correlation with telomerase activity (TA) level.


METHODS
FISH was performed on paraffin sections from six common naevi, eight Spitz naevi, 12 melanomas, six melanoma metastases and nine control samples of normal skin. Telomere mean maximum diameter (Feret max), area and number per nuclear area were calculated by image analysis on fluorescent images elaborated through KS400 and in situ imaging system (ISIS) for FISH analysis programs. Mean TA level was also calculated in all lesions and correlated with telomere parameters.


RESULTS
Telomere number per nuclear area was significantly lower in melanomas and metastases than in benign common and Spitz naevi and in control skin (7 small middle dot24 +/- 3.3; 6.11 +/- 3 vs. 14.46 +/- 5.6; 16.92 +/- 7.8; and 12.59 +/- 3.4, respectively; P < 0 .001). No significant differences were found for the other telomere parameters. In common and Spitz naevi, telomere number was positively correlated with Feret max (P = 0.046 and P < 0.0001, respectively). TA was significantly higher in melanomas and metastases than in the other groups (70.18 +/- 25.2; 105.07 +/- 30 vs. 2.16 +/- 2.4; 2 .99 +/- 2.1; 2 +/- 1.2, respectively; P< or = 0. 001) and it was inversely correlated with telomere number per nuclear area in melanomas (P = 0.0041). No other significant correlations were found.


CONCLUSIONS
Encouraging results have been obtained from quantitative telomere evaluation in the diagnosis of melanocytic lesions, although an analysis of a larger number of cases would be necessary to provide more reliable data. An extreme shortening of some telomeres probably results in the decrease of telomeric signals and the lower mean number of detectable telomeres in melanomas and metastases. In melanomas, telomere number per nuclear area is also inversely correlated with TA levels. Quantitative FISH of melanocytic lesions could give more specific information at the cellular level in telomere and telomerase fields of investigation.
Miracco
C
C
Institute of Pathological Anatomy and Histology, University of Siena, Italy. miracco@unisi.it
Margherita De Santi
M
M
Schürfeld
K
K
Santopietro
R
R
Lalinga
A V
AV
Fimiani
M
M
Biagioli
M
M
Brogi
M
M
De Felice
C
C
Luzi
P
P
Andreassi
L
L
D016022:Case-Control Studies; D006801:Humans; D007091:Image Processing, Computer-Assisted; D017404:In Situ Hybridization, Fluorescence; D008545:Melanoma / Q000201:enzymology / Q000556:secondary / Q000648:ultrastructure; D009363:Neoplasm Proteins / Q000378:metabolism; D009506:Nevus / Q000201:enzymology / Q000648:ultrastructure*; D009508:Nevus, Pigmented / Q000201:enzymology / Q000648:ultrastructure; D012878:Skin Neoplasms / Q000201:enzymology / Q000556:secondary / Q000648:ultrastructure*; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000648:ultrastructure*
D009363:Neoplasm Proteins; D019098:Telomerase