Annamaria Guazzi

Comparison of the results of immunocytochemical assays for biologic variables on preoperative fine‐needle aspirates and on surgical specimens of primary breast carcinomas

Fine‐needle aspiration biopsy (FNAB) is a well‐documented procedure for the diagnosis and biologic characterization of breast carcinoma. In order to compare the immunocytochemical expression of biologic parameters on cytology and on histology, estrogen receptor (ER) and progesterone receptor (PgR) status, p53 protein expression, and Ki67 growth fraction were evaluated on presurgical fine‐needle aspirates (FNAs) from breast carcinoma patients and on the corresponding surgical samples prior to any systemic therapy.

Accuracy of Fine Needle Aspiration Cytology in the Pathological Typing of Non-small Cell Lung Cancer

Background: Histological typing of non-small cell lung cancer (NSCLC) has an increasing clinical relevance due to the emerging differences in medical treatment between squamous and nonsquamous tumors. However, most NSCLCs are diagnosed in an advanced stage, and the diagnosis is often obtained exclusively by cytology either exfoliative or following fine needle aspiration. We investigated the accuracy of fine needle aspiration cytology (FNAC) in NSCLC typing as compared with histology. Methods: Over the period 2000–2009, 1182 transbronchial needle aspirate or transthoracic needle aspirate samples were obtained from patients with suspicious thoracic lesions. In 474 patients, a cytological diagnosis of primary NSCLC was obtained, and 186 (39%) of them (108 transbronchial needle aspirates and 78 transthoracic needle aspirates) received a parallel or subsequent histologic diagnosis on endoscopic biopsy (112) or surgery (74). Results: At cytology, 158 (85%) NSCLC cases were typed (89 adenocarcinoma and 69 squamous cell carcinoma), while 28 (15%) were classified as NSCLC not otherwise specified. At histology, 183 (98%) cases were typed (109 adenocarcinoma, 69 squamous cell carcinoma, 3 adenosquamous carcinoma, and 2 large cell carcinoma), and only 3 (2%) were classified as NSCLC not otherwise specified. Cytological and histological typing was concordant in 137 of 156 (88%) cases (K = 0.755; p < 0.001). The positive predictive value of FNAC in typing NSCLC was 92% for adenocarcinoma and 82% for squamous cell carcinoma. Conclusion: FNAC in expert hands is fairly accurate for typing NSCLC and can be regarded as an acceptable procedure for diagnostic and medical treatment planning purposes in most NSCLC cases, especially when more invasive approaches are unfeasible. In poorly differentiated and doubtful cases, the use of ancillary techniques, such as immunocytochemistry, may be required to improve the diagnostic yield.

Comparison Between Epidermal Growth Factor Receptor (EGFR) Gene Expression in Primary Non-small Cell Lung Cancer (NSCLC) and in Fine-Needle Aspirates from Distant Metastatic Sites

Purpose: Epidermal growth factor receptor (EGFR) gene copy number obtained by fluorescence in situ hybridization (FISH) has been recently found to predict treatment outcome in non-small cell lung cancer (NSCLC) patients receiving EGFR tyrosine kinase inhibitors. However, it is still unknown whether EGFR status differs in metastases compared with primary NSCLC. In all studies FISH have been performed on histologic material. The possibility to perform FISH analysis on cytologic material obtained by fine-needle aspiration from superficial and visceral metastases would allow us to know the real EGFR status avoiding invasive diagnostic procedures. Methods: EGFR gene copy number was analyzed by FISH on fine-needle aspirates obtained from 31 patients with metastatic NSCLC and the results were compared with those obtained on corresponding paraffin histologic sections from the primary tumor. Results: The feasibility of EGFR FISH on cytology was 90% (28 of 31 patients). EGFR FISH was positive in 61% (17 of 28 patients) of the metastases and in 36% (10 of 28 patients) of the primary tumors. Nine of the 28 cases (32%) were EGFR positive on both primary tumor and metastatic site and 10 (36%) were negative on both primary tumor and metastasis. Nine of the 28 cases (32%) showed discordance of primary tumor versus metastasis (McNemar test; p = 0.041). Conclusions: EGFR FISH can be reliably assessed on fine-needle aspirates obtained from NSCLC metastases. We found that EGFR gene copy number is discordant between primary NSCLC and the corresponding distant metastatic sites in a significant proportion of cases. These findings should be considered in future studies designed to elucidate the predictive role of EGFR FISH in NSCLC.

HER-2/neu amplification by fluorescence in situ hybridization in cytologic samples from distant metastatic sites of breast carcinoma

Amplification of the HER‐2/neu oncogene has been proposed as a target for antibody‐based therapies and as a predictor of chemoresponsiveness in advanced breast carcinoma. Few studies have concentrated on HER‐2/neu gene evaluation by fluorescence in situ hybridization (FISH) on distant metastatic sites and none have been performed on cytologic samples. The current study evaluated HER‐2/neu amplification by FISH on cytologic samples obtained from distant metastatic lesions of breast carcinoma to update HER‐2/neu characterization through a safe and easier procedure than biopsy.

Fine-needle aspiration technique for the concurrent immunocytochemical evaluation of multiple biologic parameters in primary breast carcinoma

SummaryFine-needle aspiration cytology has been already established as a reliable method for the diagnosis of breast cancer. Its application has been recently extended to immunocytochemical analysis of biological parameters. In the current study estrogen and progesterone receptors, Ki67 growth fraction, and p53 protein expression were immunocytochemically evaluated on the cellular material sampled by the same fine-needle aspirate used for the conventional cytologic diagnosis of malignancy. Fine-needle aspiration specimens from 100 patients with primary breast carcinoma were submitted to the immunocytochemical analysis. Twenty-eight percent were in premenopause; 23% had tumors with a diameter less than 2 cm, 59% from 2 to 5 cm, and 18% more than 5 cm; 60% had axillary nodal status negative, 34% positive, and 6% unknown. The concomitant immunocytochemical evaluation of all parameters was possible in 70% of the patients. A significant association was found between p53 overexpression and Ki67 values (p = 0.004), and between Ki67 values and progesterone receptor status (p = 0.003). No correlation was found between any parameter and clinical tumor size. Estrogen (p = 0.02) and progesterone (p = 0.04) receptor negativity and high Ki67 growth fraction (p = 0.005) were significantly associated with the clinical evidence of axillary node involvement. This study suggests that fine-needle aspiration cytology represents an effective practice for a simultaneous evaluation of multiple biologic indicators and could be useful as a preoperative procedure in patients who are candidates for neoadjuvant chemotherapy and/or endocrine therapy.

Estrogen and progesterone receptors in human meningiomas: Biochemical and immunocytochemical evaluation

BACKGROUND The observation that human meningiomas are rich in steroid hormone receptors has led to the hypothesis that their growth may be hormonally dependent. This study aims to correlate the biochemical expression of estrogen (ER) and progesterone receptors (PgR) with their nuclear immunoreactivity in a large series of meningiomas. METHODS The occurrence of ER and PgR in patients with primary untreated meningiomas was studied with a dextrancoated charcoal method (DCC) and the results were compared with those of an immunocytochemical assay (ICA). Progesterone and estrogen receptor determinations were performed on 103 and 99 meningiomas respectively using the DCC assay. Forty-six and 44 of these samples were immunocytochemically evaluated for the presence of PgR and ER respectively. RESULTS Of the 46 samples evaluated by both the methods, 89% were found PgR positive by DCC and 70% by ICA. The overall concordance between PgR-DCC and PgR-ICA was 80%. Whereas low concentrations of ER were found in 8/44 samples (18%) assayed by DCC, specific staining was never observed in any of the samples tested by ICA. CONCLUSIONS Our findings confirm that the majority of meningiomas are devoid of ER and that the biochemical evidence of PgR correlates well with the nuclear localization of progesterone receptors determined by immunocytochemistry.

Biological variables in non-small cell lung cancer: comparison between immunocytochemical determination on fine needle aspirates from surgical specimens and immunohistochemical determination on tissue sections

A number of biological and predictive markers of non-small cell lung cancer (NSCLC) have been sought, but these have so far been mainly evaluated on surgically resected specimens. Given that fine needle aspiration biopsy (FNAB) is being increasingly used in the diagnosis of NSCLC, its application could be extended to the immunocytochemical detection of biological parameters at the time of diagnosis before surgery. In order to assess the reliability of estimating biological markers on fine needle aspirates (FNAs) from NSCLC, the aim of this study was to compare Ki67 growth fraction, p53 and bcl-2 protein expression as revealed by the immuncytochemical assessment of FNAs obtained from surgical samples with the immunohistochemical results obtained from the corresponding histological sections. FNAs were performed on surgical specimens obtained from 29 NSCLC patients. Ki67, p53 and bcl-2 were cytologically and histologically evaluable in respectively 25, 27 and 19 cases. Concordance between FNAs and corresponding paraffin sections was 84% for Ki67, 93% for p53 and 95% for bcl-2. All of the specimens whose biological parameters were studied by immunocytohistochemistry also underwent flow cytometric DNA analysis of FNAs taken from fresh surgical specimens. Of the 29 cases, 22 were aneuploid and seven diploid. The S-phase fraction (SPF) was evaluable in 62% of cases. Comparison of SPF results on FNAs with Ki67 values evaluated on the corresponding histologic and cytologic specimens, revealed a significant correlation only with histology. Good reproducibility was also found in relation to the immunocytochemical results obtained on FNAs from different areas of the same tumour, showing that tumour heterogeneity does not affect the method. The concordance between the immunocytochemical and immunohistochemical results suggests that FNAB may be a reliable procedure for the biological characterization of NSCLC. Given its limited invasiveness, FNAB could be used in vivo for the preoperative assessment of biological parameters in patients with operable or metastatic NSCLC.

Comparison between ki-67 index and S-phase fraction on fine-needle aspiration samples from breast carcinoma†

Fine‐needle aspiration (FNA) biopsy has been used increasingly in the diagnosis and biologic characterization of breast carcinomas in patients who receive preoperative chemotherapy. Because proliferative activity of breast carcinoma has been shown to be of prognostic significance, the authors compared immunocytochemical Ki‐67 growth fraction and flow cytometric S‐phase fraction (SPF), both evaluated on FNA samples.

Advantages of multidetector-row CT with multiplanar reformation in guiding percutaneous lung biopsies

PurposeThis study aimed to assess the usefulness of multiplanar reformations (MPR) during multidetector-row computed tomography (MDCT)-guided percutaneous needle biopsy of lung lesions difficult to access with the guidance of the native axial images alone owing to overlying bony structures, large vessels or pleural fissures.Materials and methodsMDCT-guided transthoracic needle biopsy (TNB) was performed on 84 patients (55 men and 29 women; mean age 65 years) with suspected lung neoplasm by using a spiral MDCT scanner with the simultaneous acquisition of six slices per rotation. We determined the site of entry of the 22-gauge Chiba needle on native axial images and coronal or sagittal MPR images. We took care to ensure the shortest needle path without overlying large vessels, main bronchi, pleural fissures or bony structures; access to the lung parenchyma as perpendicular as possible to the pleural plane; and sampling of highly attenuating areas of noncalcified tissue within the lesion.ResultsDiagnostic samples were obtained in 96% of cases. In 73 patients, lesions appeared as a solid noncalcified nodule <;2 cm; 11 lesions were mass-like. In 22, the biopsy required MPR guidance owing to overlying ribs (18), fissures (2) or hilar-mediastinal location (2).ConclusionsMDCT MPR images allowed sampling of pulmonary lesions until now considered unreachable with axial MDCT guidance because of overlying bony structures (ribs, sternum and scapulae) or critical location (hilar-mediastinal, proximity to the heart or large vessels). Compared with the conventional procedure, the use of MPR images does not increase the rate of pneumothorax or the procedure time.RiassuntoObiettivoValutare l’utilità delle immagini multiplanar reformations (MPR), durante agobiopsia percutanea TC spirale multidetettore (TCMD) guidata, nelle lesioni polmonari difficilmente raggiungibili con la sola guida delle immagini assiali, in quanto coperte da strutture scheletriche, grossi vasi o scissure pleuriche.Materiali e metodiEseguita agobiopsia transtoracica (TNB, transthoracic needle biopsy) TCMD-guidata in 84 pazienti (55 maschi e 29 femmine, età media 65 anni) con lesione polmonare sospetta per neoplasia. Impiegato scanner TCMD a scansione elicoidale, con acquisizione simultanea di sei strati per ogni rotazione completa. Sulla base di immagini assiali native ed MPR sagittali o coronali è stato introdotto un ago “Chiba Point” centimetrato da 22 gauge (G) scegliendo: il tragitto più breve in cui non si sovrappongano grossi vasi, bronchi principali e lobari, scissure pleuriche e strutture scheletriche; ingresso nel parenchima polmonare il più perpendicolare possibile al piano pleurico; le zone di tessuto non calcifico più dense della lesione da sottoporre a biopsia.RisultatiNel 96% dei casi si è ottenuto un prelievo diagnostico. In 73 pazienti le lesioni sottoposte ad agobiopsia avevano caratteristiche di nodulo solido non calcifico di dimensioni inferiori ai 2 cm; 11 lesioni erano masse neoplastiche. In ventidue noduli è stato necessaria la guida bioptica con immagini MPR in quanto lesioni “coperte” da coste (18 noduli), da scissure (2 noduli) o a localizzazione ilo-mediastinica (2 noduli).ConclusioniLe immagini MPR ottenute con TCMD hanno permesso di raggiungere lesioni polmonari considerate un tempo inaccessibili con la sola guida delle immagini TCMD assiali. L’impiego della immagini MPR, inoltre, non incrementa, rispetto alla procedura convenzionale né i casi di pneumotorace (PNX) né il tempo di esecuzione del prelievo.

HER-2/neu evaluation by fluorescence in situ hybridization on destained cytologic smears from primary and metastatic breast cancer

OBJECTIVE To evaluate HER-2/neu amplification by fluorescence in situ hybridization (FISH) (HER-2/neu by FISH) on archival cytologic smears stained with May-Grünwald-Giemsa (MGG) stain. STUDY DESIGN Cytologic specimens from 69 breast cancer lesions (48 primary and 21 metastatic), stained with MGG stain for routine diagnostic cytology, were destained and subjected to HER-2/neu by FISH. Fifteen of the 69 samples were also evaluated by FISH on paired fresh smears. RESULTS HER-2/neu by FISH was successfully assayed in 25 of the 48 primary tumors and in 15 of the 21 metastatic lesions, corresponding to an overall feasibility of 58%. These cases had been archived between 1 month and 10 years prior to FISH analysis. Eight of the 25 primary and 5 of the 15 metastatic tumors were amplified. In 15 of the 40 evaluable cases, HER-2/neu was also assessed on the corresponding fresh smears: 8 tumors were amplified and 7 unamplified on both destained MGG and fresh smears. CONCLUSION HER-2/neu can be detected by FISH on routinely MGG-stained cytologic slides. This approach allows HER-2/neu evaluation whenever histologic sections or fresh cytologic material are not available. In these cases, HER-2/neu assessment on destained cytologic smears plays a role in the selection of targeted therapy.