Microsatellite Alterations and Cell-Free DNA Analysis: Could They Increase the Cytology Sensitivity in the Diagnosis of Malignant Pleural Effusion? (original) (raw)
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Microsatellite DNA analysis does not distinguish malignant from benign pleural effusions
Oncology Reports, 2007
Distinguishing malignant from benign pleural effusions using routine cytology is a common diagnostic problem. Recently, genetic alterations, including microsatellite instability (MSI) and loss of heterozygosity (LOH), have been described in malignant pleural effusions and proposed as methods improving diagnostics. The purpose of this study was to evaluate a panel of molecular markers for the detection of genetic alterations of cells in pleural effusions and to determine their diagnostic value as an additional test to cytologic examination. Pleural fluid and peripheral blood from 48 patients (36 male and 12 female, median age 71 years) were analyzed. Twenty-six patients had malignant pleural effusion, including 23 lung cancer and three metastatic non-pulmonary carcinoma. The control group consisted of 22 patients with benign pleural effusions. Only 14 malignancy-associated pleural effusions were cytology-positive for malignant cells (54%), whereas all benign pleural effusions were negative. DNA was extracted from all the samples and analysed for MSI and/or LOH using the following microsatellite markers: D3S1234, D9S171, D12S363, D17S250, D5S346 and TP53Alu, located at five chromosomal regions: 3p, 9p, 12q, 17q, 5q. Microsatellite analysis of the pleural fluid pellet exhibited genetic alterations in two neoplastic pleural fluid cases and in one inflammatory case. Two out of 26 (7.6%) patients with malignant pleural effusion showed genetic alterations. One exhibited MSI in three different micro-satellite markers (D17S250, D9S171, D3S134) and the other showed LOH in marker D3S134. One out of 22 (4.5%) patients with benign pleural effusion showed LOH in marker D3S134. In conclusion, genetic alterations at the level of microsatellite DNA, were detected only in very few cases of malignant pleural effusions, and in one case of benign pleural effusion. Thus, our data suggest that microsatellite DNA analysis does not facilitate the diagnosis of malignant pleural effusion.
Thorax, 1994
Background -Aneuploidy appears to be a highly specific marker for cancer, and measurement of cellular DNA content by flow cytometry is rapid and reliable. This study was undertaken to determine if the addition of DNA analysis improved the sensitivity of cytological diagnosis of malignancy in pleural fluid. Methods -Pleural effusions from 92 patients were studied by cytological examination and flow cytometry. Results -In 41 patients the final diagnosis was malignancy, there were 40 cases of benign effusions including 22 with pleural tuberculosis, and in the remaining 11 patients with biopsy proven cancer the presence of malignant cells was not found by cytological and histological means in the pleural fluid. Aneuploidy and cytological malignancy were found in 14 samples. There were seven cases with abnormal flow cytometry and negative cytological results. In 12 patients the cytological test results were positive but DNA analysis was normal. Thirty six samples of fluid were both diploid and cytologically negative. Of the 22 tuberculous effusions seven contained aneuploid cells. The sensitivity of DNA and cytological analysis was 51-2% and 63-4%, respectively. The specificity of DNA analysis was 74-5%. Conclusions -DNA analysis of cells in malignant pleural effusions is both less sensitive and specific than the cytological diagnosis. Flow cytometric analysis is not recommended for routine use in the diagnosis of pleural effusions. (Thorax 1994;49:692-694) Malignant disease causes, or is associated with, up to 77% of all pleural effusions.' Of particular prognostic and therapeutic significance is the presence or absence of malignant cells, which may be found either on cytological examination of pleural fluid or by histological examination of pleural tissue. In general, cytological examination of pleural fluid establishes the diagnosis more frequently than pleural biopsy, with positive rates of 25% to 87%.23 This variability depends mainly on the type of malignancy and on the skill of the cytologist. Moreover, in many patients with proven malignancy pleural effusions are caused by neoplastic involvement of the mediastinal lymph nodes rather than by direct invasion of the parietal pleura.4 Additional non-invasive diagnostic techniques would therefore be useful to identify tumour cells in pleural fluid.
Journal of Clinical Pathology, 2000
Aim-To analyse the practicability and potential assistance of static DNA cytometry performed by means of the remote quantitation server Euroquant and the internet in routine diagnostic analysis of pleural eVusions, and to outline the role of DNA cytometry on pleural eVusions in distinguishing between benign and malignant (and herein primary versus metastatic) eVusions. Materials and methods-Cytological smears of 294 pleural eVusions were stained with the Feulgen method. The DNA content of a minimum of 300 randomly chosen analysis nuclei and 30 reference nuclei (lymphocytes) was measured by internet connection to the remote quantitation server Euroquant. Cytometric features were derived from the histograms, and the time needed for case evaluation, the reliability of staining and measurement procedures, and the contribution to the final diagnosis were evaluated. Results-Only 120 of 294 pleural eVusions could be measured. The total measurement time for each specimen was 60 minutes. The guidelines of the consensus report on DNA measurements were fulfilled. Seventy eight malignant (18 mesotheliomas, 60 metastatic tumours) and 42 benign eVusions were measured. Seven of 78 malignant eVusions were euploid and none of 42 benign eVusions were aneuploid. The sensitivity and specificity were 91% and 100%, respectively, for distinguishing benign from malignant eVusions, and 95% and 100%, respectively, for discriminating between benign and malignant eVusions caused by metastatic malignant tumours.
Pathology and Oncology Research, 2021
Pathogenic molecular features gained specific significance in therapeutic decisions in lung carcinoma in the past decade. Initial and follow up genetic testing requres appropriate amounts and quality of tumor derived DNA, but tumor sampling, especially for disease monitoring is generally limited. Further to the peripheral blood (PB), samples from pleural fluid, accumulating in diverse lung processes might serve as an alternative source for cell-free DNA (cfDNA) for genetic profiling. In our study, cfDNA isolated from the pleural effusion and from the PB, and genomic DNA (gDNA) obtained from tissue/cellular samples were analyzed and compared from altogether 65 patients with pulmonary disease, including 36 lung adenocarcinomas. The quantity of effusion cfDNA yield appeared to be significantly higher compared to that from simultaneously collected PB plasma (23.2 vs. 4.8 ng/μl, p < 0.05). Gene mutations could be safely demonstrated from the effusion cfDNA fraction obtained from adeno...
Can We Improve the Cytologic Examination of Malignant Pleural Effusions Using Molecular Analysis?
The Annals of Thoracic Surgery, 2005
Background. Currently, 40% of patients remain undiagnosed after routine cytologic examination for malignant pleural effusions. Deoxyribonucleic acid (DNA) methylation is a robust strategy for detecting cancer early in tissue. We hypothesized that DNA methylation would be more sensitive in diagnosing patients with malignant pleural effusions than cytology.
BMC Cancer, 2012
Background: The diagnosis of malignant pleural effusions (MPE) is often clinically challenging, especially if the cytology is negative for malignancy. DNA integrity index has been reported to be a marker of malignancy. The aim of this study was to evaluate the utility of pleural fluid DNA integrity index in the diagnosis of MPE. Methods: We studied 75 pleural fluid and matched serum samples from consecutive subjects. Pleural fluid and serum ALU DNA repeats [115bp, 247bp and 247bp/115bp ratio (DNA integrity index)] were assessed by real-time quantitative PCR. Pleural fluid and serum mesothelin levels were quantified using ELISA. Results: Based on clinico-pathological evaluation, 52 subjects had MPE (including 16 mesotheliomas) and 23 had benign effusions. Pleural fluid DNA integrity index was higher in MPE compared with benign effusions (1.2 vs. 0.8; p<0.001). Cytology had a sensitivity of 55% in diagnosing MPE. If cytology and pleural fluid DNA integrity index were considered together, they exhibited 81% sensitivity and 87% specificity in distinguishing benign and malignant effusions. In cytology-negative pleural effusions (35 MPE and 28 benign effusions), elevated pleural fluid DNA integrity index had an 81% positive predictive value in detecting MPEs. In the detection of mesothelioma, at a specificity of 90%, pleural fluid DNA integrity index had similar sensitivity to pleural fluid and serum mesothelin (75% each respectively). Conclusion: Pleural fluid DNA integrity index is a promising diagnostic biomarker for identification of MPEs, including mesothelioma. This biomarker may be particularly useful in cases of MPE where pleural aspirate cytology is negative, and could guide the decision to undertake more invasive definitive testing. A prospective validation study is being undertaken to validate our findings and test the clinical utility of this biomarker for altering clinical practice.
Cancer Biology & Therapy, 2005
Fluids of body cavities result in a series of pathophysiological events associated with non-malignant and malignant conditions that lead to the formation of exudative effusion. Diagnosis of effusion from the patients is frequently troublesome for the cytologist because of the differentiation and biological behavior of different cells type in effusion. In the present study, chromosomal aneuploidy status in effusion cells derived from 32 patients including 14 patients with non-malignant diseases and 18 patients with malignant diseases [including malign mesothelioma (n = 6), adeno carcinoma (n = 10), small cell carcinoma (n = 2)] was analyzed by using fluorescence in situ hybridization (FISH) with centromere specific probes for chromosomes 9 and 11. There was significant difference in the incidence of chromosomal 9 and 11 aneuploidies when compared with controls (P = 0.000). However, aneuploidies of chromosomes 9 and 11 in effusion cells from patients with malignant disease had significantly higher than in effusion cells from patients with non-malignant (P = 0.000), suggesting that chromosomes 9 and 11 are frequently involved in the status of disease. The present study indicates that there is a association between chromosomes aneuploidies and pleural effusion cell status. Chromosome aneuploidies in non-malignant group may be an indicator of premalignancy.