EGFR mutation testing on cytological and histological samples in non-small cell lung cancer: a Polish, single institution study and systematic review of European incidence.

The targeted treatment of advanced non-small-cell lung cancer (NSCLC) depends on confirmation of activating somatic EGFR mutation. The aim of the study was to evaluate the incidence of EGFR mutations in NSCLC detected in cytological and histological material and present literature review on European EGFR mutation incidence. 273 patients with confirmed NSCLC were entered into the study: 189 histological, paraffin-embedded materials, 12 fresh and 72 fixed cytological specimens. DNA was extracted from both types of material and the EGFR mutation in exons 18-21 was analyzed by direct sequencing. In addition the EGFR gene copy number in cases with sufficient histological material (110 patients) was evaluated by fluorescent in situ hybridization (FISH) technique. The percentage of EGFR somatic mutations was 10.62%. FISH positive results (amplification or high polysomy of EGFR gene) were identified in 33 patients (30.0%). The strongest clinicopathological correlation with the EGFR mutation was found for histological type (adenocarcinoma; p < 0.01), gender (females; p < 0.01) and FISH positive result (p < 0.05). This is the first, single institution study that estimates the EGFR mutation incidence in the Polish population. Cytological material recovered from fixed preparations and stained with hematoxylin and eosin showed DNA quality comparable to fresh tumor cells and histological samples.

Endobronchial metastasis from hepatocellular carcinoma - a case description with literature review.

Endobronchial metastases from hepatocellular carcinoma are very rare. Up to date, no more than 7 cases were reported. The authors present a case of 20-year old female with metastatic hepatocellular carcinoma to superior lobar bronchus. Examination of cytological and small biopsy specimens obtained from bronchoscopy revealed characteristic microscopic features and immunohistochemical profile of hepatocellular carcinoma.

Double, synchronous pituitary adenomas causing acromegaly and Cushing's disease. A case report and review of literature.

Double pituitary adenomas are very rare and present up to 1 % of pituitary adenomas in unselected autopsy series and up to 2 % in large surgical series. We report a case of a 47-year-old man presented slight clinical features of acromegaly with 2 years duration. Endocrine evaluation confirmed active acromegaly and revealed adrenocorticotropin hormone-dependent hypercortisolemia. Preoperative magnetic resonance imaging of the pituitary demonstrated clearly separated double microadenomas with different intensity. The patient underwent transsphenoidal surgery and both tumors were completely removed and were fixed separately. The histological and ultrastructural examination confirmed coincidence of the double, clearly separated pituitary adenomas in one gland. Postoperative function of the hypothalamo-hypophyseal axis was normalized. We conclude from this case and a literature review that double endocrinologically active pituitary adenomas leading to acromegaly and Cushing's disease may occur. Additionally, a review of the literature regarding multiple pituitary adenomas has also been performed.

Reproducibility of immunohistochemical scoring for epidermal growth factor receptor expression in non-small cell lung cancer: round robin test.

CONTEXT: The addition of cetuximab to first-line chemotherapy substantially prolonged survival in patients with advanced non-small cell lung cancer whose tumors expressed high levels of epidermal growth factor receptor (EGFR; immunohistochemistry score of ≥200 on a scale of 0-300). OBJECTIVE: To evaluate the interobserver reproducibility of this EGFR immunohistochemistry scoring system, based on both the tumor cell membrane staining intensity (graded 0-3+) and the percentage of cells staining at each intensity. DESIGN: In parts 1 (initial feasibility study) and 2 of this 2-part round robin test, sections of different non-small cell lung cancer tissue microarrays were stained in a central reference laboratory. Following reference evaluation, EGFR expression in 30 selected tumor cores was characterized in serial sections by lung cancer pathology specialists. The reproducibility of scoring by different raters was assessed. Analysis of between-rater agreement was based on the allocation of EGFR immunohistochemistry scores into low- (<200) and high- (≥200) EGFR expression groups. RESULTS: After discussion with raters of the issues impacting reproducibility identified in part 1 and following adjustment of processes, part 2 of the round robin test showed a high interobserver agreement in EGFR immunohistochemistry scoring, with an overall concordance rate of 90.9% and a mean κ coefficient of 0.812. Specimens with a reference EGFR immunohistochemistry score of lower than 200 and of 200 or higher showed mean concordance rates of 94.7% and 85.6%, respectively. CONCLUSIONS: After appropriate training, assessing EGFR expression by this immunohistochemistry-based method allowed a highly reproducible allocation of non-small cell lung cancers into clinically relevant high- or low-EGFR expression groups.

The characteristics of the sentinel lymph node metastasis in predicting the axillary lymph node status in patients with breast carcinoma.

INTRODUCTION: Lymph node metastases are the most significant prognostic factors in patients with breast carcinoma. A positive sentinel lymph node (SLN) biopsy is followed by an axillary lymph node (ALN) dissection. In sentinel lymph node negative cases the risk of positive non-sentinel ALN is very low though not absent. The aim of this study was to determine predictive factors for non-sentinel lymph node metastases on the basis of sentinel lymph node metastasis characteristics as well as features of the primary tumour. MATERIAL AND METHODS: 128 patients who had a positive SLN biopsy for breast carcinoma in 2005-2007 were identified. The breast carcinoma metastases in each SLN were assessed according to their location within the node (subcapsular, mixed subcapsular and parenchymal, parenchymal, multifocal or extensive) and metastatic infiltration of perinodal tissue was also reported. These data were correlated with the ALN involvement and characteristics of the primary tumour. RESULTS: The strong predictors of the ALN metastasis included the SLN metastasis diameter (7.6 vs. 4.4 mm) and size classified according to WHO classification (ITC 0 vs. 100%, micrometastasis 23.5 vs. 76.5%, macrometastasis 51.9 vs. 48.1%). The SLN metastases with a diameter of above 3 mm were associated with approximately twice more frequent ALN metastases. In an extensive location of SLN metastasis the highest percentage of ALN metastases was found (65 vs. 35%). The weak predictors of ALN metastases were: primary tumor diameter (> 2 cm), immunohistochemical HER2 positive status, infiltration of sentinel perinodal tissue by metastasis, histological primary tumour grade. CONCLUSIONS: Some additional details, which can be easily evaluated in a routine SLN examination in breast carcinoma, have a predictive value of the ALN metastatic status and should be included in the histopathological report.

HER2 status in breast cancer determined by IHC and FISH: comparison of the results.

HER2 (human epidermal growth factor receptor 2) status became an important prognostic and predictive factor in breast carcinoma clinical management. There are two main techniques of evaluation of HER2 status: immunohistochemistry (IHC) for the protein expression and fluorescence in situ hybridization (FISH) for amplification of HER2 gene. The aim of the study was to compare the results obtained by IHC and FISH methods in determination of HER2 status in breast cancer. Three hundred and sixty breast cancer specimens were examined. Patients were operated in the Oncology Centre in Warsaw. IHC and FISH were performed in every case. IHC was performed with DAKO HercepTest and FISH with Oncor-QBiogene reagents. IHC results were classed into 4 groups, accordingly to the four-tier DAKO criteria system (0, 1+, 2+, 3+). FISH results were divided into three main categories: NA--no amplification, LA--low amplification and HA--high amplification. The number of copies of chromosome 17 was also assessed. Over 90% of cases described by IHC as 3+ exhibited amplification of HER2/neu gene. Remaining cases were positive with IHC, but presented no gene amplification. This might be due to the subjective assessment of the membrane staining. Another possibility is that overexpression of the protein was caused by mRNA stability or disorders in receptor degradation. The majority of cases classed by IHC as 2+ were also negative by FISH (80%). One fifth of IHC 2+ tumours were found to exhibit gene amplification. Remaining cases showed no amplification of HER2/neu gene, combined with aneuploidy of chromosome 17. All cases described by IHC as 0/1+ were also HER2-negative by FISH. IHC is well-established method of assessing HER2 status in breast cancer. Nonetheless, a group of cases described as 2+ should be additionally examined using FISH. The results obtained by the latter method are more reliable. In order to improve accuracy and gain the highest quality of HER2 status evaluation, in 2+ cases both methods should be applied.