Sabato 27 ottobre 2012 - Pacini Editore

274
The specificity of the FNC was 98.2, the false positive rate was
of 8.3%, the false malignant value was of 29% and 25% using
V600-BRAF.
Discussion. In patients with unquestionable cytological evidences
of PTC, the preoperative detection of the V600E BRAF
mutation may have a direct impact on treatment. Recently, Xing 4
suggested that, for prognostic purpose, perhaps all patients with
cytologically diagnosed PTC should be preoperatively tested for
BRAF mutation. In this respect, this test provides information
that are additional and non redundant to those provided by a well
taken and correctly interpreted thyroid FNA.
With an one-time withdrawal thyroid (FNC), we obtained cytological
and molecular data that may be useful both for diagnosis
and prognosis of the patient. We consider important to obtain
additional molecular information with a single withdrawal from
thyroid nodules to avoid unnecessary steps how to call the patient
with all the problems inherent in it. We keep the material in suspension
and carry out cytological research BRAF V600 mutation.
In our study eleven FNC had a conclusive cytological evidence of
PTC (Tir5), four FNC had a souspicius cytological of PTC (Tir4),
fifteen FNC had an undeterminate diagnosis (Tir3) and 227 not
neoplastic (Tir2).
Regardless of the sample collection methodology, all FNC were
suitable for molecular analysis and were tested with a sequence
of thyroglobulin and the amount of extracted DNA was more
than one hundred nanograms, deemed sufficient quantities; in
three cases Tir5 and two cases Tir4 V600 BRAF mutation was
detected. In all remaining cases classified Tir2, V600E BRAF
mutation was not detected.
Our data showed that, in a routine clinical setting, FNC specimens
can be handled properly to provide both morphological and
molecular information. Our study focusing on the single steps
required to aliquot the aspirated material into routine smears
and DNA extraction buffer may help to implement BRAF testing
in the prognostic evaluation of PTC diagnosed by FNA. Our
proposed method ensures that this test does not interfere with
conventional cytology diagnostic accuracy.
references
1 Dean DS, Gharib H. Epidemiology of thyroid nodules. Best Pract Res
Clin Endocrinol Metab 2008;22:901-11.
2 Davies H, Bignell GR, Cox C, Stephens P, et al. 2002 Mutations of the
BRAF gene in human cancer. Nature 417:949-54.
3 Cohen Y, Xing M, Mambo E, et al. BRAF mutation in papillary thyroid
carcinoma. J Natl Cancer Inst 2003;95:625-7.
4 Xing M, Westra WH, Tufano RP, et al. BRAF mutation predicts poorer
clinical prognosis for papillary thyroid cancer. J Clin Endocrinol Metab
2005;90:6373-9.
5 Troncone G, Cozzolino I, Fedele M, et al. Preparation of Thyroid FNA
Material for Routine Cytology and BRAF Testing: A Validation Study.
Diagnostic Cytopathology2009;38(3).
6 Hassell LA, Gillies EM, Dunn ST. Cytologic and molecular diagnosis
of thyroid cancers: Is it Time for Routine. Cancer Cytopathol 2011:
10.1002/.20186.
Sensitivity and positive predictive value
of HMB-45 and MArT-1 as immunocytochemical
markers of recurrent/metastatic melanoma on
fine needle cytology samples: an experience with
250 cases in 5 years (2007-2012)
F. Fulciniti1 , A. Galzerano2 , A. Cipolletta Campanile1 , A. Gioioso1 ,
F. Caccavello1 1 S.S.D. di Citopatologia, S.C. di Anatomia Patologica e Citopatologia,
Istituto Nazionale Tumori “Fondazione G. Pascale”, Naples, Italy; 2 Pathology
Unit, Centro Hospitalar Lisboa Ocidental, Lisbon, Portugal
Background. Metastatic melanoma may be difficult to distinguish
from other neoplasms such as carcinomas, lymphomas or
sarcomas due to its highly variable morphologic picture. Even
though the various cytomorphologic presentations of melanoma
CONGRESSO aNNualE di aNatOmia patOlOGiCa SiapEC – iap • fiRENzE, 25-27 OttOBRE 2012
on Fine Needle Cytology (FNC) samples are well known to
cytopathologists, a diagnosis of recurrent/metastatic melanoma
should always be validated by appropriate ancillary techniques,
due to its usually unfavorable prognostic and medico-legal implications.
Immunocytochemical stains for melanocytic markers
on FNC samples should couple the highest possible diagnostic
accuracy with the least possible number of immunocytochemical
stains on a given sample to be both accurate and cost-effective.
Materials and methods. We retrospectively analyzed 250 cases
of recurrent/metastatic melanoma diagnosed on FNC samples
from various body sites between January 2007 and May 2012
in Our Institution. There were 99 female- and 151 male patients
(F/M ratio 1/ 1.5), with a median age of 59 yrs. All of the cytological
diagnoses were confirmed by tissue study and clinicopathological
follow-up. For each patient, one representative
routine, wet-fixed Papanicolaou-stained smear was dismounted
and stained for HMB 45 (Novocastra, clone HMB 45) by a semiautomated
immunostainer (Leica, Bond Max). Positive cases
showed a diffuse, usually intense, granular cytoplasmic positivity
in > 10% of the neoplastic cells. In cases with negative HMB45
staining with cytomorphologic features suggestive for recurrent/
metastatic melanoma, a supplementary wet-fixed Papanicolaoustained
smear was dismounted and stained for MART-1 (Melan-
A, Novocastra, Clone A 103). MART-1 staining was generally
more diffuse (> 30% of the neoplastic cells) than HMB 45 in 34
cases in which both immunostains were performed on the same
case (13.6 %). HMB-45 and MART-1 staining results were used
to determine the sensitivity and positive predictive value of immunocytochemistry.
Results. Of 250 cases, 231 were positive for HMB-45, with a
sensitivity of 92,4%, and 19 were negative (7.6%), while of 34
cases in which we performed MART-1, 27 were positive, with
a corresponding sensitivity of 79%. 15 cases were positive for
HMB45 and MART1. The combined sensitivity HMB-45+ and/
or MART-1+ was 98%. The positive predictive value of a positive
HMB-45 stain and/or and positive MART-1 stain was 100%.
Both HMB45 and MART-1 were negative in 5 cases (2%). The
negativity was correlated to cellular necrosis in 1 case, to a diagnosis
of desmoplastic melanoma in 2 cases (which were double
negative also on histology) and to the amelanic epithelioid phenotype
in 2 more cases.
Conclusion. HMB-45 and MART-1 demonstrated to be very
sensitive and cost-effective to confirm the cytomorphological
diagnosis of recurrent/ metastatic melanoma on FNC samples.
We suggest the routine usage of HMB-45 or both HMB-45 and
MART-1 versus the usage of extended immunocytochemical
panels. HMB45 staining seems to have a superior outcome when
used on alcohol fixed smears as opposed to formalin-fixed cells
blocks. Microphthalmia transcription factor (MITF) or other alternative
markers for melanoma should be tested on desmoplastic
or other variants of melanoma which are double negative for
HMB45 and MART-1-.
references
1 Fetsch PA, Marincola FM, Filie A, et al. Melanoma-associated antigen
recognized by T cells (MART-1): the advent of a preferred immunocytochemical
antibody for the diagnosis of metastatic malignant
melanoma with fine-needle aspiration. Cancer.1999;87:37-42.
2 Sheffield MV, Yee H, Dorvault CC, et al. Comparison of five antibodies
as markers in the diagnosis of melanoma in cytologic preparations.
Am J Clin Pathol 2002;118:930-6.
3 Dorvault CC, Weilbaecher KN, Yee H, et al. Microphthalmia transcription
factor: a sensitive and specific marker for malignant melanoma
in cytologic specimens. Cancer 2001;93:337-43.
4 Murali R, Thompson JF, Uren RF, et al. Fine-needle biopsy of metastatic
melanoma: clinical use and new applications. Lancet Oncol
2010;11:391-400