In the existing study a comparative analysis of keratin typing and DNA contented was carried out in human lung tumors from transthoracic fine needle aspiration biopsies (TFNAB) (18 patients) or from surgically resect tumor tissues (14 patients).

In the existing study a comparative analysis of keratin typing and DNA contented was carried out in human lung tumors from transthoracic fine needle aspiration biopsies (TFNAB) (18 patients) or from surgically resect tumor tissues (14 patients). According to the cytologic and histologic features, 2 of the 32 tumors were diagnosed as benign tumors, 11 as squamous enclosed space carcinomas, 12 as adenocarcinomas, and 7 as undifferentiated large solitary abode; squalid carcinomas. Two cases in the adenocarcinoma and common in the undifferentiated large enclosed space carcinoma groups were pulmonary metastasis or secondary primary tumors. Malignant small rooms of tumors which reacted positively with KK860 anticytokeratin polypeptides No. 10 and 11 (and hence contain keratinizing cells) displayed diploid DNA make contented in a flow cytometric assay regardless of their cytologic or histologic appearance. In contrast, all tumors which lacked similar positive cells (most of which were defined as adenocarcinomas and undifferentiated tumors) were hyperdiploid. The choke correlation between high DNA satisfaction and both malignancy and the absence of advanced squamous differentiation (keratinization) glance ats that such combined analysis may provide of recent origin tools for the cytologic diagnosis and prognosis of lung cancers.

The diagnosis and classification of human tumors comprise crucial degrees for an accurate prognosis and selection of optimal therapeutic procedures

For lung tumors, a universal diagnostic approach is the cytologic examination of transthoracic fine needle aspiration biopsy (TFNAB) specimens.[1-5] While this examination frequently provides valuable information on the nature of the tumor, it is frequently handicapped by limited morphology of distinct cellular manner of makings and the inherent absence of distinctive histotypic-morphologic features.[6,7]

A more precise differential diagnosis of TFNAB specimens may thus be obtained through specific immunocytologic examination of like tumors using a variety of markers in the same state [i]or[/i] condition as cytokeratin polypeptides as described in our previous study[8] The issues of that study indicated that a significant proportion of lung tumors originally diagnosed according to cytology as squamous cell carcinomas, are in fact, adenocarcinomas or mixed adeno-squamous tumors. Thus, cytokeratin typing, using various monoclonal antibodies (MoAb), provides more precise information onward the origin of tumors than popularly attainable by cytologic examination only

In an attempt to obtain additional information forward TFNAB samples of lung tumors, which may bear forward their biological properties, we have carried without a comparative analysis of keratin immunocytologic typing and DNA appease analysis in the same specimens. This approach was based forward the rationale that the clinical and biological behavior of tumors (including step of malignancy and aggressiveness) may be related to the state of proliferation of the tumor lonely dwellings Flow cytometric analysis of propidium-iodide stained nuclei provides quantitative information in succession cell ploidy and the percentage of solitary abode; squalids in the different phases of the mitotic cycle[910] The reported comes suggest that in contrast to benign tumors and normal tissues, malignant lonely dwellings especially adenocarcinomas and undifferentiated tumors, contain significantly higher evens of DNA.[11-14]

MATERIALS AND METHODS

The diagnosis of lung cancer (total 32 patients) was made either from TFNAB material (18 patients), or from surgically resect tumor tissue (14 patients). According to the cytologic and histologic features, 2 of the tumors were diagnosed as benign tumors, 11 as squamous small room carcinomas, 12 as adenocarcinomas, and 7 as undifferentiated large confined apartment carcinomas. Two cases from the adenocarcinoma clump and one from the undifferentiated large solitary abode; squalid carcinoma group were pulmonary metastasis or inferior primary tumors. Of the 32 patients, 29 were men and 3 women with an average age of 626 years (Table 1)

Sample Collection and Preparation

Surgical specimens of the tumor tissue were snap-frozen in liquid nitrogen-cooled isopentane and kept at -70 [degrees] C Single small cavity suspensions of tumor tissues were obtained by way of mincing the specimen with scissors and forcing it between the walls of wire mesh.[15] Cells were maintained in RPMI 1640 medium (Biolab, Jerusalem, Israel). To obtain a pure representation of the tumor, relatively large parts of tissue were processed.

[TABULAR DATA OMITTED]

The TFNAB was done using a 5-inch, 22-gauge needle Again to obtain a veracious representation of the tumor enclosed spaces the needle was moved inside the tumor between the walls of the depth several times before taking it not at home enabling multiple sampling. The majority of solitary abode; squalids recovered from the needle were used for Papanicolaou staining[16] and light microscopic evaluation while the caesura were suspended in RPMI 1640 medium and used for DNA analysis or keratin typing.

DNA Analysis

Propidium-iodide staining of nuclei was carried not at home according to the procedure bring to maturityed by Vindelov et al,[17] and the fluorescence of about 10000 nuclei was measured for specimen by flow cytometer (FACS 440 Becton Dickinson). The data were analyzed according to the system described by Dean.[18] As normal diploid rule specimens, human lymphocytes obtained from normal donors were used. The DNA index (DI) for each specimen was calculated from the ratio of the mean value of [Gsub0]/[Gsub1] peak in the patient sample and in the govern diploid cells. Diploid tumor populations were defined as having a single [Gsub0]/[Gsub1] peak at the same range as the [Gsub0]/[Gsub1] peak for the sway cells, DI = 1.0 [+ or -] 01 (01 is twice the standard deviation of the mean [Gsub0]/[Gsub1] peak of the direct diploid lymphocytes). Tumors were considered hyperdiploid if there was a [Gsub0]/[Gsub1] peak with DI value higher than 11

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