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BC and GC samples and corresponding lymph node Fleiss-Cohen weighted K statistics was used to
metastases. The study was approved by review board assess the concordance rate between HER2 status of
of University of Messina. the primary carcinomas and metastatic synchronous
Methods lesions. K values between 0 and 0.2 were regarded as
This cohort contained 127 surgical BC and GC no agreement, between 0.21 and 0.4 as fair agreement,
specimens, together with the corresponding regional between 0.41 and 0.6 as moderate agreement,
synchronous metastatic lymph nodes. In brief, 65 primary between 0.61 and 0.8 as substantial agreement, and
BC and 62 primary GC (male:female = 39:88; age ranged between 0.81 and 1 as almost perfect agreement. The
between 44 and 95 years with mean age of 69.32 years) statistical association between HER2 status and the
were retrospectively collected from the archive of the other histopathological parameters was assessed using
Department of Human Pathology at the University Chi-squared test. P < 0.05 was considered statistically
of Messina. No patients had received neo-adjuvant signifi cant. All statistical analyses were performed
chemotherapy or other therapies before surgery. using the SPSS package version 6.1.3 (SPSS, Chicago,
IL, USA).
The primary GC was classifi ed for localization and
histology type according to WHO 2010, Lauren’s Results
classifi cation and HER2 status of the tumor were Thirty GC cases (48.40%) were localized in the lower
available for all cases. Similarly, histology, grade, third of the stomach, 22 (35.48%) in the middle
hormone receptor status, Ki-67, and HER2 status were third and 10 (16.12%) in the upper-third (four of
recorded for all BC cases. Patient identifi cation was which were localized at gastro-esophageal junction).
not disclosed in this publication, and all patients had Thirty-fi ve GC cases (56.45%) were diagnosed
provided written consent to their medical information histopathologically according to the WHO criteria as
being used for research purposes, according with the adenocarcinoma (tubular, papillary, tubulo-papillary,
Helsinki declaration.
and mucinous), 20 cases (32.25%) as poorly cohesive
For each case, 3 m thick tissue sections from carcinoma, and 7 cases (11.30%) were mixed both.
two different formalin-fi xed paraffi n-embedded According to Lauren’s classifi cation, 35 cases (56.45%)
representative tissue blocks of the primary tumor and were classifi ed as intestinal type, 20 cases (32.25%)
metastatic lymph nodes (at least four for each case) as diffuse and 7 cases (11.30%) as mixed. Thirty-two
were prepared and immunohistochemical stained for of these 62 primary GC (51.61%) were recorded
HER2 expression. In brief, the immunohistochemistry as low-grade tumors, while 30 cases were high
was carried out by using a DAKO HercepTest grade (48.39%). HER2 immunohistochemical staining
™
kit (Dako, Glostrup, Denmark) with an automated showed that 11 primary GCs (17.74%) were scored for
procedure (DAKO Autostainer Link 48) according 3+ HER2 expression, while 4 cases were 2+ (6.42%),
to manufacturer’s instructions. Antigen retrieval was 5 cases 1+ (8.10%), and 42 cases (67.74%) were not
performed by 3 cycles in 0.01 mol/L citrate buffer expressed HER2 at all. FISH analysis revealed no
pH 6.0 in a microwave oven at 750 W. For HER2 score amplifi cation in all of these cases with HER2 scores
was used to semiquantitatively assess HER2 expression of 2+ or more. Taken together, in primary GC, HER2
level, that is, for the primary GC, 0, absent staining; 1+, was overexpressed in 11 cases (17.74%) but there was
faint and discontinuous membranous staining in < 10% no HER2 amplifi cation in 51 cases (82.26%). The
of neoplastic elements; 2+, light to moderate lateral, overall concordance rate of HER2 status in primary
baso-lateral or complete membranous staining in > GC between corresponding synchronous metastases
10% of neoplastic elements; 3+, strong, intense lateral, was 90.32%, whereas a change in HER2 status
baso-lateral or complete staining in > 10% of neoplastic was observed in 6 (9.68%) [ Table 1], e.g. 4 cases
elements and for BC, 3+ score was defi ned when strong with HER2 amplifi cation in the primary GC but no
membranous staining was noted in at least 30% cells, amplifi cation in the metastasized tumors [negative
2+ when weak to moderate complete membranous conversion; Figure 1a and b], two of these discordant
staining was evidenced in 10-30% of tumors cells, cases did not show HER2 amplifi cations in the
1+ when a faint or weak and incomplete membrane primitive tumor but amplifi ed in the lymph node
staining was observed and 0 when no staining was metastases [positive conversion; Figure 1c and d and
observed or when staining was present in < 10% of Table 2].
neoplastic cells.
In the primary BC, the most frequent histology type was
Furthermore, fl uorescence in situ hybridization (FISH) ductal invasive carcinomas with the following grading:
was performed using a HER2 FISH PharmDx 4 G1 (6.25%), 28 G2 (43%), and 33 G3 (50.75%).
™
kit (Dako) in those cases with HER2 immunostaining HER2 overexpression occurred in 14 (21.53%) of
score for 2+ or more. HER2 amplifi cation was recorded primary BC, 4 (6.15%) of which exhibited a score 2+,
when HER2 to CEP17 signal ratio was > 2.0. 2 (3.09%) a score 1+, while 45 (69.23%) cases didn’t

22 Journal of Cancer Metastasis and Treatment ¦ Volume 1 ¦ Issue 1 ¦ April 15, 2015 ¦