Material and Method
In our study, laryngectomy materials examined between January 2010 and
December 2016 were investigated retrospectively. 57 cases who were
diagnosed with LSCC and have not previously received chemotherapy and
radiotherapy were included in the study. The clinical information of the
patients was obtained from the automation system of our hospital.
Pathology reports, slides and paraffin blocks of the patients were
extracted from our archive. 4-micron sections were taken from the blocks
where the tumor was the densest and were put into the Roche Ventana
automatic immunohistochemistry staining device ( Ventana Roche,
ABD) after the tissues placed on charged slides in a 70-degree
drying oven for 15 minutes. The tissues in the device were treated with
ULTRA Cell Conditioning Solution, hydrogen peroxide, HSPA-2 antibodies
(Nova Castra, Leica, Newcastle, United Kingdom) after being subjected to
deparaffinization and dehydration processes respectively.
The immunohistochemical stains were evaluated by two pathologists.
Nuclear and/or cytoplasmic staining was considered positive for HSPA-2.
For HSPA-2, staining rate=0 (Grade-0), without staining 1-10% = 1
(Grade-1), 11-49% = 2 (Grade-2), ≥50% = 3 (Grade-3) were accepted.
HSPA-2 was evaluated as follows; staining intensity: 0; no staining
(Grade-0), 1+; weak cytoplasmic and nuclear staining (Grade-1), 2+;
moderate cytoplasmic and nuclear staining (Grade-2), 3+; strong
cytoplasmic and nuclear staining (Grade-3) (Figure 1). The
immunoreactivity score (histoscore) was calculated using the method of
multiplying the intensity and rate. The histoscore values for HSPA-2 0
were evaluated as follows; negative (Grade-0), 1-3; weak (Grade-1), 4-6;
moderate (Grade-2), 7-9 (Grade-3); strong (13). The connection between
HSPA-2 staining rate, intensity, histoscore and age, gender and
important prognostic parameters such as histological grade, the TNM
stage, the Primary Tumor, lymph nodule metastasis, the presence of
lymphovascular invasion, survival rate was examined.
Our study was approved by our local ethics committee (08-31/26.12.2019).