Correlation between morphology and human telomerase gene amplification in bronchial brushing cells for the diagnosis of lung cancer

The aim of this study was to investigate the frequency of amplification of the human telomerase gene (TERC), as measured by fluorescence in situ hybridization (FISH), in routine liquid‐based cytological preparations from bronchial brushing specimens, and to assess the associations between TERC amplification, cytological diagnosis, and cytological morphology, in order to obtain further insight into these associations. Bronchial brushings from 102 patients with lung carcinoma (52 squamous‐cell carcinomas, 22 adenocarcinomas, 28 small cell lung carcinomas) and 40 patients with nonmalignant disease were used. Amplification of TERC was performed using a commercially available two‐color FISH probe, and slides were prepared for the SurePath liquid‐based Pap test (LPT) using the same samples. Amplification of TERC was significantly associated with histological diagnoses (P < 0.05). Patients with lung cancer, and especially those with nonsmall cell lung cancer, had significantly higher percentages of cells with amplification of TERC than did patients with nonmalignant disease (P < 0.05). Comparing the FISH and LPT results, there was no significant difference in diagnostic sensitivity between the two methods (P > 0.05). However the difference in diagnostic sensitivity of the two methods for squamous‐cell carcinoma was significant (P < 0.01). FISH can be performed on bronchial brushing specimens to detect amplification of TERC. This test may be an adjunct to cytology screening, especially in squamous‐cell carcinoma, and may provide an indication of the potential of individual lesions to progress. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.     

Respiratory cytology in the era of molecular diagnostics: a review

Carcinoma of the lungs remains one of the primary causes of cancer mortality in the United States and represents a significant diagnostic challenge. Current diagnostic protocols depend substantially on cytology as an initial diagnostic modality. Pulmonary cytology can be diagnostically challenging with false positive and false negative diagnoses being relatively frequent. False positive diagnoses remain a significant problem for the cytologist with benign conditions including reactive atypia of type II pneumocytes, reactive bronchial respiratory epithelium, basal cell hyperplasia, and reactive metaplastic squamous cells being potentially misinterpreted as carcinoma. False negative diagnoses also occur usually attributable to sampling. Traditionally, cytopathologists were expected to recognize carcinoma when present and subdivide it into small cell or nonsmall cell varieties. With the advent of targeted therapy, expectations now include separation of adenocarcinoma from squamous cell carcinoma. Additionally, molecular testing for EGFR mutations and ALK rearrangements is now required as an accompaniment to morphologic diagnosis. This review summarizes the morphologic appearances of the common and diagnostically important carcinomas of the lung and discusses diagnostic pitfalls responsible for false positive and false negative diagnoses. Molecular testing for selection of targeted therapy is also reviewed.     

The diagnostic value of endobronchial ultrasound‐guided needle biopsy in lung cancer and mediastinal adenopathy

Endobronchial ultrasonography (EBUS) has emerged as a new diagnostic tool that allows the bronchoscopist to see beyond the airway, including pulmonary and mediastinal lesion. The real time EBUS‐guided transbronchial needle aspiration (TBNA) has advanced the diagnostic yield in primary lung pathology and mediastinal lymph node staging of lung carcinoma. Sixty‐four patients (36 males, 28 females, ages ranging from 16 to 86 years) with peribronchial lung lesions and mediastinal and/or hilar lymph node lesions underwent EBUS‐TBNA. All patients had intraoperative cytological assessment by smears on aspiration samples or touch preparation on needle core biopsies. The cytological final diagnoses were categorized as negative, suspicious/positive, and non‐diagnostic. Forty‐nine samples were obtained from lymph node lesions and 15 samples were obtained from lung lesions. In cytology specimens, 32 patients had suspicious/positive diagnoses and 32 patients had negative diagnosis. In follow‐up histology specimens, 35 patients had malignant diagnoses, including 18 adenocarcinomas, 8 small cell carcinomas, 6 squamous cell carcinomas, 1 metastatic hepatocellular carcinoma, 1 metastatic melanoma, and 1 lymphoma. Twenty‐nine patients had negative diagnoses. Sensitivity and specificity were 88.9% and 96.4%, respectively. Positive and negative predictive values were 97.0% and 87.1%, respectively. Diagnostic accuracy was 92.2%. EBUS‐TBNA is an efficient and effective technique for diagnosis of intrapulmonary and mediastinal/hilar lymph nodes. It becomes significantly invaluable on clinical management for staging in those patients with lung cancer of other metastatic malignancies. This technique enables us to obtain tissue samples for quick diagnoses beyond central airway with minimal complications. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.     

Accuracy of morphological diagnosis of lung cancer in a department of respiratory medicine.

In view of the importance of determination of cell type in patients with bronchial carcinoma who are to be given chemotherapy a series of 140 histologically proved cases was analysed to assess the accuracy of pretreatment diagnosis by cytology and bronchial biopsy. Final histological diagnosis was made after thoracotomy (131 cases) or necropsy (9 cases). Before treatment, malignancy was diagnosed in 71% of cases with 91% accuracy in cell typing. Diagnostic yield was greatest in squamous carcinoma (81%) and least with adenocarcinoma (50%). Large-celled and small-celled anaplastic carcinomas gave yields of 61% and 68% respectively. Accuracy of cell typing was high (90-97%) except for large-celled anaplastic carcinoma (71%). All diagnostic methods were not employed in every case. In patients supplying samples the diagnosis was made by sputum cytology in 57%, by rigid bronchoscopy cytology in 57%, by fibreoptic bronchoscopy cytology in 71% and biopsy in 53%. Causes of diagnostic failures and possible means of increasing accuracy of pretreatment diagnosis were discussed.     

Diagnostic accuracy of cytology and biopsy in primary bronchial carcinoma.

The accuracy of diagnosis in 656 patients with the four common histopathological types of primary lung cancer has been assessed by comparing the cell type diagnosis made on cytological and histological investigation with that determined by examination of the surgically resected or necroscopy specimen. The accuracy of diagnosis achieved by cytological examination of sputum and bronchial aspirate, and by bronchial biopsy histology was over 85%. The least accurate diagnostic procedure was percutaneous needle biopsy (62%). Squamous and small cell tumours were accurately diagnosed by all four investigations but errors were made in the diagnosis of large cell and adenocarcinomas. Nearly half the number of patients (43%) with large cell carcinoma were later reclassified as having squamous carcinoma and of the patients with adenocarcinoma 32% had been predicted to be squamous and 18% large cell carcinoma. We consider such quality control of pretreatment diagnosis mandatory in management of individual patients and before enrollment in clinical trials.     

Adenosquamous carcinoma of the lung diagnosed by cytology?: A diagnostic dilemma

Adenosquamous cell carcinomas of the lung are rare tumours and are associated with a poor prognosis compared to other non-small cell carcinomas. We report a case of a solitary lung carcinoma evaluated by bronchial brush and lavage cytology, bronchial biopsy and pleural fluid cytology. Cytological assessment of the pleural fluid demonstrated non-small cell carcinoma and immunohistochemical staining confirmed a metastatic lung adenocarcinoma. The bronchial brush and lavage specimens, however, demonstrated the cytomorphological features of squamous cell carcinoma, which was confirmed by the bronchial biopsy. The finding of a mixed squamous and glandular component predicts a poor prognosis for this patient. The identification of a squamous component with the non-small cell carcinoma is important as this excludes the patient from anti-VEGF monoclonal antibody treatment due to the increased risk of haemorrhage.     

Non-small cell lung carcinomas with neuroendocrine features. A light microscopic, immunohistochemical and ultrastructural study of 11 cases

Eleven resected primary lung carcinomas classified as large cell carcinomas or squamous cell carcinomas, but showing some microscopic resemblances to bronchial carcinoid and small cell carcinoma, were studied. All cases were neurone-specific enolase and protein gene product 9.5 positive, indicating neuroendocrine differentiation. Staining for bombesin, C-terminal peptide of human pro-bombesin and chromogranin was positive in some cases. Electron microscopy showed dense-core granules in six of seven cases investigated, the remaining case showing small granules of uncertain nature. All but one patient died within 15 months after operation. These data indicate that neuroendocrine differentiation in non-small cell carcinomas of the lung may in some cases be suspected on routine histology. The follow-up data suggest that the identification of these cases might have implications for prognosis and therapy, and consequently for diagnostic lung tumour classification.     

HnRNP A2/B1和P53蛋白对肺癌诊断价值的探讨

目的：采用流式细胞术检测肺癌患者支气管肺泡灌洗液（BALF）脱落细胞中HnRNP A2/B1和外周血单个核细胞中P53蛋白水平,探讨联检HnRNP A2/B1和P53蛋白在肺癌诊断中应用价值。方法：收集30例肺癌患者的支气管肺泡灌洗液（BALF）的脱落细胞和其外周血,用流式细胞术检测脱落细胞表达HnRNP A2/B1和外周血中的P53蛋白水平;同时检测30例肺部良性疾病患者为对照组。结果：肺癌患者的BALF脱落细胞表达HnRNP A2/B1的含量明显高于对照组（P〈0.01）;非小细胞肺癌患者中HnRNP A2/B1明显高于小细胞肺癌（P〈0.05）。NSCK临床TNM分期Ⅰ-Ⅱ期的患者与Ⅲ-Ⅳ期相比,HnRNP A2/B1无显著性差异（P〉0.05）。肺癌患者外周血中P53蛋白明显异于对照组（P〈0.01）;小细胞肺癌患者的P53蛋白明显高于非小细胞肺癌（P〈0.01）。结论：FCM检测肺癌患者BALF脱落细胞中HnRNP A2/B1和外周血P53,有助于肺癌的早期诊断。     

Follow-up of abnormal gynecologic cytology: a college of American pathologists Q-probes study of 16132 cases from 306 laboratories

OBJECTIVES: To measure the percentage of women with abnormal gynecologic cytology who have follow-up within 1 year and to identify patient and laboratory characteristics associated with higher percentages of follow-up. DESIGN AND SETTING: Retrospective identification of patients with abnormal cervicovaginal cytology and identification of the initial clinical follow-up activity during the 12 months following the cytologic diagnosis. MAIN OUTCOME MEASURE: Percentage of women receiving follow-up. RESULTS: Three hundred six laboratories reported follow-up information on 16 132 patients with gynecologic cytology diagnoses of carcinoma, high-grade squamous intraepithelial lesion, low-grade squamous intraepithelial lesion, or glandular intraepithelial lesion. The following percentages of women received follow-up within 1 year: 85.6% of patients with cytologic diagnoses of carcinoma, 87.2% with diagnoses of high-grade squamous intraepithelial lesion, 82.7% with diagnoses of low-grade squamous intraepithelial lesion, and 84.9% with diagnoses of glandular intraepithelial lesion. Within 6 months, 82.2% of patients with cytologic diagnoses of carcinoma, 82.4% with diagnoses of high-grade squamous intraepithelial lesion, 71.9% with diagnoses of low-grade squamous intraepithelial lesion, and 74.7% with diagnoses of glandular intra-epithelial lesion received follow-up. Overall, 90. 8% of patients who received follow-up within the 1-year time frame of this study had their follow-up completed within 6 months. Specific follow-up activities and their frequencies are listed for each diagnostic category. Patients 30 years old or younger and pregnant patients had lower follow-up percentages. CONCLUSIONS: With less than 83% of patients with high-grade squamous intraepithelial lesion or carcinoma cytology findings having available documentation of follow-up within 6 months, and less than 88% within 1 year, there is room for improvement in this area of health care. Monitoring and critical analysis of the follow-up process is a starting point for improvement.     

OV-TL 12/30 (keratin 7 antibody) is a marker of glandular differentiation in lung cancer

The immunoreactivity of OV-TL 12/30, a monoclonal antibody to keratin 7 was investigated on paraffin-embedded human lung cancer tissues of 61 patients. A modified AEC-immunoperoxidase method with pepsin pre-digestion was used. In normal lung tissue keratin 7 was found in bronchial and bronchiolar epithelium, pneumocytes and compound glands. Squamous metaplasia of the bronchial tree was negative. All 24 squamous cell carcinomas were negative irrespective of grade of differentiation. All differentiation grades of 20 adenocarcinomas including bronchioalveolar carcinomas were positive. Since six large cell anaplastic carcinomas did not react with keratin 7 antibody these tumours are considered to be of squamous cell rather than adenocarcinomatous origin. Small cell anaplastic carcinomas were negative in 10 of 11 cases. Our study demonstrates that this keratin 7 antibody is useful in differentiating between squamous cell carcinoma and adenocarcinoma of the lung and it may be particularly useful in making the correct diagnosis in small lung biopsy specimens.     

The relevance of ultrastructural examination in the classification of primary lung tumours

Biopsies from 50 primary lung tumours were classified according to the World Health Organisation's Histological Typing of Lung Tumours. They were also subjected to electron microscopic examination. Comparison of the diagnoses made by these separate methods showed that many poorly differentiated squamous cell carcinomas had been incorrectly classified. Agreement was good in the diagnosis of adenocarcinoma whilst ultrastructural examination of small anaplastic carcinomas disclosed a neuroendocrine tumour with a combination of squamous and glandular elements. Large cell anaplastic carcinoma proved to be a 'waste-basket' containing tumours which displayed ultrastructural characteristics of poorly differentiated squamous cell carcinoma, poorly differentiated adenocarcinoma or neuroendocrine carcinoma. Electron microscopy was also valuable in characterization of other pulmonary tumours whose identity could not be resolved at the light microscopic level. Ultrastructural examination may provide a better understanding of the histogenesis and derivation of lung tumours, as well as their behaviour and therapeutic response.     

A high-grade malignancy bronchial mucoepidermoid carcinoma with features of giant cell carcinoma

A high-grade mucoepidermoid carcinoma associated with giant cell carcinoma of the bronchus was found in a 41-year-old man. Light and electron microscopic examinations showed glandular and squamous elements. The histology of the giant cell carcinoma was similar to those of previously reported giant cell carcinomas. It was assumed that the giant cell carcinoma in the present case is the result of transformation of mucoepidermoid carcinoma originating in the bronchial gland.     

34betaE12 Cytokeratin Immunodetection in the Differential Diagnosis of Small Cell Tumors of Lung

The group of small cell tumors of the lung includes fine following: (1) small cell carcinoma (SCC) of neuroendocrine (NE) origin, (2) poorly differentiated squamous carcinoma, (3) the rare basaloid (basal cell) carcinomas, and (4) malignant lymphomas, primitive neuroectodermal tumors (PNETs), and rhabdomyosarcomas. The differential diagnosis among these entities carries a heavy therapeutic impact but may be difficult in small biopsy specimens or in cytologic material, especially if necrosis or artifactual alterations are present. The use of additional techniques such as immunostaining for NE markers is not always helpful, since immunoreactive chromogranin A is detectable in only a small percentage of small cell carcinomas. It has recently been reported that in the aerodigestive tract 34betaE12 cytokeratin (CK) immunostaining selectively labels non-NE carcinomas, including squamous cell carcinoma, adenocarcinoma, and the rare basaloid carcinoma. We evaluated the role of such CK immunodetection in the differential diagnosis of small cell lung tumors in cytologic and biopsy specimens. Eighty-one lung tumors diagnosed by means of endoscopic bronchial biopsy, fine needle aspirate, or bronchial washing were collected. They included 43 small cell NE carcinomas and 38 cases used as controls (comprehensive of 2 large cell neuroendocrine carcinomas, 4 carcinoid tumors, 30 cases of non-NE lung carcinomas, 2 cases of bronchial infiltration by non-Hodgkin lymphomas). 34betaE12 CK immunoreactivity was found in 29/30 cases of non-NE carcinomas, but in only 3/43 SCCs. The latter showed positivity in only a few scattered cells. The 2 cases of bronchial infiltration by malignant lymphoma as well as the 4 cases of carcinoid tumors and the 2 cases of large cell neuroendocrine carcinomas were negative. These findings were confirmed in the surgical specimens of operatedon cases. We conclude that, in lung carcinoma biopsies showing a small cell pattern, presence of 34betaE12 CK immunoreactivity favors a non-NE carcinoma, whereas its absence supports the diagnosis of SCC. Int J Surg Pathol 8(4):317-322, 2000     

A HIGH-GRADE MALIGNANCY BRONCHIAL MUCOEPIDERMOID CARCINOMA WITH FEATURES OF GIANT CELL CARCINOMA

A high-grade mucoepidermoid carcinoma associated with giant cell carcinoma of the bronchus was found in a 41-year-old man. Light and electron microscopic examinations showed glandular and squamous elements. The histology of the giant cell carcinoma was similar to those of previously reported giant cell carcinomas. It was assumed that the giant cell carcinoma in the present case is the result of transformation of mucoepidermoid carcinoma originating in the bronchial gland.     

Rinse fluid and imprint smear cytology of bronchial biopsies in diagnosis of lung tumors

The usefulness of rinse fluid and imprint smear cytology of the bronchial biopsy has been studied in diagnosis of lung cancer. However, scarce data is available regarding rinse fluid cytology of biopsy. The aim of this study was to evaluate these cytologic techniques for their diagnostic accuracy. Bronchial biopsy was taken in 52 patients clinically/radiologically suspected to have lung carcinoma. Imprint smears of the biopsy were prepared, following which it was put in balanced saline solution to collect rinse fluid of biopsy before transferring it to formalin for fixation. Cytological diagnosis from imprint and rinse fluid smears was compared with histopathological diagnosis. Malignancy was detected in 45 cases of 52 patients on histopathology. Positive result was given by rinse fluid cytology in 34 (65.4%) cases while diagnostic accuracy was 78.8%. The imprint smears were positive for malignancy in 44 (84.6%) cases with diagnostic accuracy of 98.08%. There were no false-positive results, but one case was incorrectly typed by both the techniques. Imprint smear cytology has a better diagnostic accuracy and efficacy over rinse fluid while the two cytologic techniques can be used in combination routinely with biopsy to provide an early and reliable diagnosis in lung cancer.     

Primary pulmonary adenoid cystic carcinoma: report of a case diagnosed by fine-needle aspiration cytology

Adenoid cystic carcinoma of the lower respiratory tract is an uncommon tumor that can arise in the mainstem bronchus and often presents as an endobronchial mass lesion causing bronchial obstruction with post obstructive atelectasis and pneumonia. Exfoliative cytology is seldom useful in the diagnosis of primary bronchial adenoid cystic carcinoma, because these neoplasms usually have a submucosal location with often intact mucosa. Since most endobronchial adenoid cystic carcinomas are endoscopically visible, bronchoscope-guided fine-needle aspiration constitutes an excellent approach to establish a pathologic diagnosis. The fine-needle aspiration cytology of primary pulmonary adenoid cystic carcinoma has been rarely described. We report a case of primary adenoid cystic carcinoma of the lung having characteristic cytologic features and correlate with computed tomography, bronchoscopic, and histological findings. Bronchoscope-guided aspiration cytology provided a conclusive diagnosis of adenoid cystic carcinoma, which was further corroborated by histology in the pneumonectomy specimen. Diagn. Cytopathol. 2004;30:51-56.     

P63 in pulmonary epithelium,pulmonary squamous neoplasms,and other pulmonary tumors

p63 is a p53-homologous nuclear protein that appears to play a crucial role in regulation of stem cell commitment in squamous and other epithelia. In this study, p63 expression was examined in benign lung and in neoplasms of pulmonary origin. Eighty sections from routinely fixed and processed archival bronchoscopic biopsy or lobectomy specimens were pretreated with citric acid (pH 6.0) for antigen retrieval, then incubated overnight with anti-p63 monoclonal antibody 4A4. Slides were stained using a streptavidin-biotin kit and diaminobenzidine as chromagen, and were counterstained with hematoxylin. In normal lung, p63 intensely stained nuclei of bronchial reserve cells but did not stain ciliated cells, alveolar epithelial cells, or nonepithelial cells. The lower strata of squamous metaplastic bronchial epithelium stained positively. All squamous-cell carcinomas stained positively (n = 30). In some well-differentiated carcinomas, staining was found at the periphery of tumor nests but was negative in central zones showing squamous maturation. Poorly differentiated carcinomas showed very high proportions (80% to 100%) of p63-positive nuclei. All small-cell carcinomas were p63 negative (n = 9). Staining of bronchioloalveolar carcinomas (n = 7) and adenocarcinomas (n = 23) was variable: some tumors showed no detectable staining, others showed heterogeneously positive staining. Adenosquamous carcinomas (n = 5) displayed a unique basalar staining pattern. Carcinoid tumors were almost entirely negative (n = 5). We conclude that p63 is expressed in benign bronchial stem cells, in neoplastic cells with either squamous differentiation or squamous differentiating potential, and in a subpopulation of adenocarcinomas. p63 immunostaining may also aid in some histopathologic distinctions, such as in small biopsies where the differential diagnosis is poorly differentiated squamous carcinoma versus small-cell carcinoma. A stem cell biology-based classification system for squamous carcinomas is proposed.     

Cytological typing of primary lung cancer: study of 100 cases with autopsy confirmation.

In order to assess the accuracy of bronchial aspiration cytology in typing lung cancer, tissue sections from 100 autopsy cases of lung cancer were compared with the cytology features observed in the same patients prior to death. There was 100% accuracy in the cytology of small-cell carcinoma; 90% in squamous-cell carcinoma; 70% in adenocarcinoma; and 50% in undifferentiated large-cell carcinoma. The observed discrepancies probably reflect intrinsic tumor properties rather than problems attributable to either the bronchial aspiration method or cytology interpretation, especially in cases involving advanced lung carcinoma. Because the highest accuracy rate was in detecting small-cell carcinoma, it is recommended that only the distinction between small-cell and non-small-cell forms be made on cytologic grounds and that further categorizations only be rendered in cases with unquestionable cytomorphological features.