The value of epithelial membrane antigen expression in separating benign mesothelial proliferation from malignant mesothelioma: a comparative study

Differentiating reactive mesothelial (RM) proliferation from malignant mesothelioma (MM) can be cytologically challenging. There have been discordant studies reporting the value of epithelial membrane antigen (EMA) in differentiating RM from MM. In this study, we investigated the expression of two different clones of EMA in RM and MM. Twenty cases of pleural effusion smears of RM and 20 cases of MM with their corresponding cell blocks were retrieved from the hospital computer system. Diagnosis of MM was confirmed by surgical decortication or pneumonectomy with immunostaining studies and/or electron microscopy. Cases of RM were confirmed by clinical history and histology. Cell blocks were formalin-fixed, paraffin-embedded, and immunostained for EMA clone Mc5 and EMA clone E29. The positive rates for clone Mc5 were 14/20 (70%) for MM and 12/20 (60%) for RM and EMA clone E29 were 15/20 (75%) for MM and 0/20 (0%) for RM. The sensitivity and specificity for EMA clone Mc5 were 70 and 40%, respectively. For EMA clone E29, the sensitivity and specificity were 75 and 100%, respectively. In conclusion, both RM and MM immunostained for EMA clone Mc5, indicating that it is not a reliable immunocytochemical marker for differentiating RM from MM. EMA clone E29 was negative in all cases of RM and positive in 75% of MM and therefore is a reliable immunocytochemical marker for differentiating RM from MM.     

Scoring system for differential diagnosis of malignant mesothelioma and reactive mesothelial cells on cytology specimens

Cytology is the only useful tool in the detection of malignant mesothelioma (MM) at an early stage. No other methods, such as immunocytochemistry or electron microscopy, are available to distinguish MM from reactive mesothelial cells (RMC). Some objective analysis of cytology specimens is necessary. On the basis of our case review and cytological features described in previous articles, we developed a scoring system for malignant mesothelioma (SSMM) of effusion cytology to distinguish MM cells from RMC. Mesothelioma cells in effusions from 22 patients (20 pleural and 2 peritoneal mesotheliomas) were compared with RMC from 20 patients without obvious tumor cells and 50 effusions containing metastatic carcinoma cells. The SSMM is based on characteristic features of mesothelial and malignant cells. The total achievable score is 10 points: one point each is given for variety of cell size, cyanophilic cytoplasm with villosity/windows/bleb, sheet‐like arrangement, mirror‐ball‐like cell clusters, nuclear atypia, and cannibalism, respectively. Further two points each are ascribed for acidophilic large nucleoli and multinucleated cells with more than eight nuclei. The total score for each of the 22 mesotheliomas was more than 5 points. On the other hand, all RMC and the 50 metastatic carcinoma cases scored less than 3 points, aside from two cases that were treated with OK432. No single characteristic feature was observed to be consistent within the 22 mesotheliomas analyzed. Ancillary use of immunocytochemistry, such as podoplanin (D2‐40) and calretinin, supported the diagnostic accuracy of the SSMM. SSMM is useful for the differential diagnosis of MM. Diagn. Cytopathol. 2009. © 2009 Wiley‐Liss, Inc.     

Cytopathologic differential diagnosis of malignant mesothelioma, adenocarcinoma and reactive mesothelial cells: A logistic regression analysis

Distinguishing malignant mesothelioma, adenocarcinoma and reactive mesothelial proliferation in both cytologic and surgical pathologic specimens is often a diagnostic challenge. Conventional cytomorphologic assessment is an important step in the differential diagnosis of these entities.The pleural effusion cytologies from 40 cases of malignant mesothelioma, 40 cases of adenocarcinoma and 30 cases of reactive mesothelial proliferation diagnosed between 1997 and 2007 were reviewed. Twenty-seven cytologic features which are regarded as useful in the differential diagnosis of mesothelioma, adenocarcinoma and benign mesothelial proliferation were assessed. These cytologic features were subjected to a stepwise logistic regression analysis. Three features were selected to distinguish malignant mesothelioma from adenocarcinoma: giant atypical mesothelial cell (P = 0.0001), nuclear pleomorphism (P = 0.0001) and acinar structures (P = 0.0001), the latter two being characteristics of adenocarcinoma. The variables selected to differentiate malignant mesothelioma from reactive mesothelial cells were: cell ball formation (P = 0.0001), cell in cell engulfment (P = 0.0001) and monolayer cell groups (P = 0.0001), the latter being a feature of benign mesothelial proliferation. When these selected variables were subjected to a stepwise logistic regression analysis, the logistic model correctly predicted 90% of cases of benign mesothelial proliferation versus 97.5% of malignant mesothelioma and 92.5% of malignant mesothelioma versus 92.5% of adenocarcinoma.Conventional cytomorphologic assessment is the first step to establish an accurate diagnosis in pleural effusions. Several cytologic features have predictive value to separate malignant mesothelioma from adenocarcinoma and reactive mesothelial proliferation.     

Membranous HEG1 expression is a useful marker in the differential diagnosis of epithelioid and biphasic malignant mesothelioma versus carcinomas

Sialylated HEG1 has been reported as a highly specific and sensitive mesothelioma marker but a comprehensive evaluation of its expression in carcinomas in different organs, various sarcomas and reactive mesothelial proliferations has not been reported. The aim of this study was to evaluate the clinical applicability of HEG1 as a marker in the diagnosis of mesothelioma. HEG1 immunoreactivity was evaluated in whole sections of 122 mesotheliomas, 75 pulmonary carcinomas, 55 other carcinomas, 16 mesenchymal tumors, and 24 reactive mesothelial proliferations and in tissue microarrays containing 70 epithelioid (EM), 36 biphasic (BM), and 2 sarcomatoid mesotheliomas (SM). In whole sections and tissue microarrays, respectively, membranous HEG1 was expressed in 93.0% and 85.5% of EM, 81.3% and 69.4% of BM, 0% and 0% of SM. HEG1 was not expressed in pulmonary adenocarcinomas. HEG1 was expressed as cytoplasmic immunoreactivity in pulmonary squamous cell carcinomas (21.7%). Membranous HEG1 staining was seen in ovarian carcinomas (66.7%), thyroid carcinomas (100%), reactive conditions (16.7%), and mesenchymal tumors (18.8%). The sensitivity of membranous HEG1 expression to distinguish EM/BM from all carcinomas was 88.8%. The specificity for the differential diagnosis between EM/BM and all carcinomas and pulmonary carcinomas was 92.3% and 98.7%, respectively.     

Assessment of cell cycle state may facilitate the histopathological diagnosis of malignant mesothelioma

AIMS: The differentiation of benign pleural conditions from malignant mesothelioma may be difficult, especially with a small biopsy. We have tested the hypothesis that assessment of the cell cycle status is of value in the histopathological diagnosis of such biopsies, by comparing 33 malignant mesotheliomas with 36 cases of reactive mesothelial hyperplasia and reactive pleural fibrosis. METHODS AND RESULTS: Biopsies were investigated for proliferative status by immunostaining for a novel antibody, MCM2, all of which showed nuclear expression of MCM2 at higher frequency than Ki67 (P < 0.0001). Counts in areas of maximum tumour staining showed significantly higher labelling indices (LIMax) in epithelioid and sarcomatoid mesotheliomas compared with reactive mesothelial hyperplasia and reactive pleural fibrosis (P < 0.0001 for both). Average counts (LIAve) revealed a significant increase in epithelioid mesothelioma compared with reactive mesothelial hyperplasia (P < 0.0001). CONCLUSION: We consider MCM2 to be a useful adjunct in the differential diagnosis of malignant mesothelioma.     

Telomerase activity and proliferation index in aggressive mature B-cell lymphoma: comparison to germinal center phenotypic markers

Cell proliferation may be evaluated by various methods, including Ki-67 immunohistochemistry and measures of telomerase activity. Both methods would theoretically show comparable increases in a given case. To evaluate the relationship between these 2 markers of proliferation in aggressive mature B-cell lymphomas, 48 cases were studied. The study group included 5 cases of mantle cell lymphoma (MCL); 6 cases of Burkitt's/Burkitt's-like lymphoma (BL); 9 cases of follicular lymphoma, grade 3 (FLC); and 28 cases of diffuse large B-cell lymphoma (DLC). Telomerase activity was measured as total product generated (TPG) units, and TPG results for the aforementioned cases were compared to the TPG results for 10 cases of reactive follicular hyperplasia. An overlap in TPG scores between reactive cases and lymphoma cases was found. Significant differences in both log TPG (P = 0.0443) and Ki-67 (P = 0.0006) were seen in the different lymphoma types. A positive correlation between Ki-67 percentage and TPG score was identified in FLC (r = 0.9281; P = 0.0003), but a poor correlation between these 2 indicators was seen in the other lymphoma types. Cluster analysis identified distinct patterns for MCL, FLC, and BL, but heterogeneous patterns for DLC. Because increases in both Ki-67 proliferation and telomerase activity are reported in normal germinal centers (GCs), these tests were also evaluated for usefulness as markers of a GC cell phenotype. Among the FLC and DLC cases, features of a GC phenotype significantly correlated with increased Ki-67 percentage (P = 0.0152), but not with increased log TPG. An elevated log TPG correlated with CD10 expression, and elevated Ki-67 percentage correlated with both CD10 and BCL-6 expression. TPG level and Ki-67 percentage did not correlate with the presence of t(14;18) or BCL-2 protein expression. Although the proliferation patterns were fairly distinctive for MCL, FLC, and BL, these studies show that markers of cell proliferation do not by themselves,identify distinct subtypes of large cell lymphomas. With the exception of FLC, the tumors exhibited poor correlation between telomerase activity and Ki-67 proliferation index. These tests did show some correlation with expression of GC cell phenotypic markers, however.     

乳腺良恶性病变组织端粒长度和端粒酶活性检测

目的　比较乳腺良恶性病变端粒长度改变及其在肿瘤发生发展中的意义 ;探讨端粒酶活性与临床病理参数的关系及其在乳腺癌诊断中的价值。方法　Southern印迹杂交检测TRF长度 ,端粒重复扩增分析 (TRAP)方法检测端粒酶活性。结果　乳腺癌组织平均TRF为 (5 2± 2 8)kb ,与正常组织比较明显缩短 (P <0 0 0 1) ,从正常乳腺组织到乳腺良性病变、乳腺原位癌及浸润性癌平均TRF呈递减趋势。 5 8例乳腺癌中 4 9例端粒酶阳性 (84 7% ) ,端粒酶活性与临床病理参数无相关性 ;癌旁组织端粒酶为阴性 ,而 7例乳腺增生症和 6例乳腺纤维腺瘤中分别有 1例端粒酶阳性 ,与乳腺癌比其差异有显著性 (P <0 0 0 1)。结论　端粒长度在肿瘤发生发展过程中渐进性缩短 ,并最终触发端粒酶的激活 ;端粒酶活性检测有望成为乳腺癌诊断的可靠标记物     

肿瘤标志物联合检测对良、恶性腹水鉴别诊断价值

目的探讨联合测定血清和腹水中3种肿瘤标志物CEA、CA19-9和CA50水平及其腹水/血清比值(F/S)对良、恶性腹水鉴别诊断的价值。方法明确诊断的腹水患者180例,其中男性76例,女性104例;年龄25～78岁,平均年龄54.99岁。61例良性和119例恶性腹水患者同时采血和抽取腹水,采用电化学发光法测定CEA、CA19-9和CA50。结果腹水肿瘤标志物的水平高于血清正常上限值(CEA>5μg/L、CA19-9>37kU/L、CA50>20mg/L)时,良性腹水的F/S比值都<1.2,而恶性腹水的绝大多数F/S比值>1.2。腹水CEA、CA19-9和CA50的上述上限值结合F/S比值>1.2,诊断灵敏度分别为63.9%、59.6%和74.6%,皆显著高于单纯的腹水浓度和F/S比值的灵敏度(P<0.01)。3项联合诊断灵敏度达93.8%,特异度100%。结论与单纯血清或腹水中浓度测定相比,同时测定肿瘤标志物CEA、CA19-9和CA50的腹水浓度和F/S比值能显著提高诊断的灵敏度和特异度,对良、恶性腹水的鉴别诊断具有较大的价值。     

Sensitivity and specificity of immunohistochemical antibodies used to distinguish between benign and malignant pleural disease: a systematic review of published reports

AIMS: A systematic review of published reports that have evaluated the ability of immunohistochemistry and argyrophil nucleolar organizing region (AgNOR) staining to distinguish between benign and malignant pleural disease. METHODS: Nineteen relevant papers published during the period 1979-2005 were identified. Individual results of immunohistochemistry for five diagnostic antibodies were extracted to calculate diagnostic sensitivity and specificity. results from five of these studies that had evaluated proliferation markers or AgNOR staining techniques were also summarized. RESULTS: Most antibodies demonstrated poor to moderate diagnostic ability. Desmin and epithelial membrane antigen (EMA) were the most useful, with sensitivity and specificity both above 74%. The combination of EMA and AgNOR was reported as having 95% diagnostic sensitivity. A high MCM2 labelling index also differentiated between benign and malignant pleural disease. CONCLUSIONS: Immunohistochemistry is of limited value, but newer diagnostic methods may be useful additions in this area of pathology. The diagnostic importance of histological features seen on plain tissue sections is emphasized as vital for correctly differentiating between benign pleural disease and malignant pleural mesothelioma.     

Characterization of endoglin and Ki-67 expression in endothelial cells from benign and malignant lesions of the uterine cervix

Activation of endothelial cells is often associated with the cellular proliferation in vitro . CD105 is a more specific marker of activated endothelial cells from tumor vessels and Ki-67 is used to assess the proliferation status of both tumor and endothelial cells. The aim of the present study was to evaluate the status of endothelial cells using CD105 and Ki-67 immunohistochemistry in benign and malignant lesions of the uterine cervix. Double stain for CD105/Ki-67 in benign and malignant lesions of the uterine cervix showed that these two markers had divergent expression on endothelial cells from associated tumor blood vessels dependent on lesion type and proliferation status of tumor cells. Absence of CD105/Ki-67 coexpression in endothelial cells was correlated with histopathology of the uterine cervix lesions and tumor proliferative status. The present findings suggest that CD105 expression is an early event, specific for premalignant lesions of the uterine cervix, while endothelial proliferation assessed on Ki-67 combined with the lack of CD105 expression is often associated with invasive cervical carcinoma.     

P57和Ki-67在葡萄胎鉴别诊断中的作用

目的:探讨P57和Ki-67蛋白在完全性和部分性葡萄胎鉴别诊断中的作用.方法:分别收集正常胎盘绒毛、部分性葡萄胎和完全性葡萄胎石蜡标本各12例,应用免疫组织化学方法检测P57和Ki-67蛋白在这些病变中的分布及表达水平.结果:P57蛋白在正常绒毛及部分性葡萄胎组织中主要分布于绒毛的细胞滋养叶细胞及间质细胞,两组间阳性率...     

亚砷酸对CNE1细胞株增殖和端粒酶hTRT的影响

目的　观察亚砷酸对人鼻咽癌CNE1细胞株增殖的抑制作用及端粒酶逆转录酶 (hTRT)的影响。方法　人鼻咽癌细胞株CNE1用不同浓度亚砷酸处理 ,MTT法观察不同浓度亚砷酸作用不同时间后对CNE1细胞生长的影响 ,流式细胞仪检测不同条件作用后CNE1细胞增殖指数和凋亡指数及周期分布改变 ,免疫组化法检测增殖细胞核抗原 (PCNA)及端粒酶hTRT表达情况。结果　亚砷酸抑制人鼻咽癌细胞株CNE1细胞的增殖 ,其作用随着亚砷酸浓度增加和作用时间延长而增加 ;一定浓度亚砷酸作用于CNE1细胞后 ,其S期细胞比例及增殖指数随着时间的延长而逐渐减少 ,但凋亡指数逐渐增多。随着亚砷酸浓度增加 ,PCNA及hTRT阳性细胞平均灰度值逐渐下降 (P <0 0 5 )。结论　亚砷酸可抑制人鼻咽癌细胞株CNE1的生长 ,除了诱导凋亡以外 ,调控DNA合成及细胞增殖的PCNA蛋白合成降低 ,抑制端粒酶的活性可能是亚砷酸的部分抗肿瘤作用机制。     

Comparison of three cytologic preparation methods and immunocytochemistries to distinguish adenocarcinoma cells from reactive mesothelial cells in serous effusion

We assessed whether a panel of seven antibodies is useful in the differentiation of adenocarcinoma cells (ACCs) from reactive mesothelial cells (RMCs) in effusion samples and to determine optimal specimen preparation conditions for immunocytochemical analysis of effusion samples. Immunocytochemistry (ICC) was performed on three types of effusion preparations from the same effusion specimens: ethanol-fixed smears, ethanol-fixed cell-blocks, and formalin-fixed cell-blocks. Commercially available antibodies MOC-31, Ber-EP4, CA19-9, CEA, EMA, CA125, and HBME-1 were tested on RMCs from four samples of various etiology and 15 samples of adenocarcinoma from various primary sites. Ethanol-fixed smears showed strong immunoreactivity to all antibodies tested. The immunoreactivity of ethanol-fixed and formalin-fixed cell-blocks was significantly lower with all antibodies except CA19-9. Smear preparations are more sensitive than cell-blocks for immunocytochemical study. A panel of antibodies MOC-31, Ber-EP4, CA19-9, and CEA appears to be suitable to distinguish between ACCs and RMCs.     

超声造影增强模式在乳腺良恶性肿块鉴别诊断中的价值

目的探讨超声造影增强模式对乳腺良恶性肿块鉴别诊断的价值。方法选择40例经病理组织检查证实的乳腺肿瘤患者,均为女性,年龄25～54岁,平均年龄35岁。其中良性肿块25例,恶性肿块15例。使用Siemens Sequoia 512彩色多普勒超声诊断仪,造影剂采用SonoVue,行超声造影增强方式检查。结果乳腺良恶性肿块超声增强方式不同,良性肿块以整体不同程度地均匀型增强改变为主(80%,20/25),恶性肿块主要表现为不均匀增强或周边增强(86.7%,13/15)。结论乳腺良恶性肿块超声造影增强模式不同,可为鉴别诊断提供帮助。     

p53 immunostaining in the distinction between benign and malignant mesothelial proliferations using formalin-fixed paraffin sections.

The distinction between reactive mesothelium and mesothelioma in pleural biopsy specimens is notoriously difficult, and conventional immunohistochemical markers have provided no relief. The object of this study was to examine the frequency of immunohistochemically detectable p53 overexpression in routinely processed, paraffin-embedded tissue from pleural mesotheliomas and from pleura showing reactive mesothelial hyperplasia, using a polyclonal antibody to formalin-resistant p53 epitopes, and to consider the diagnostic utility of this antibody in the distinction between mesothelioma and reactive mesothelium in pleural biopsy specimens. Immunostaining was enhanced by pepsin predigestion prior to the application of the primary antibody. Positivity occurred in 10/16 epithelial mesotheliomas, 9/19 biphasic mesotheliomas, 2/12 sarcomatous mesotheliomas but in none of 20 reactive pleura. Immunostaining was particularly intense in some of the biopsy specimens, which may be due to the rapidity with which these small pieces of tissue were fixed. In conclusion, this study suggests that p53 immunostaining can help to distinguish epithelial or biphasic mesothelioma from reactive mesothelial hyperplasia in formalin-fixed, paraffin-embedded pleural biopsy specimens.     

Expression of PC-cell-derived growth factor in benign and malignant human breast epithelium

PC-cell-derived growth factor (PCDGF, progranulin) is a novel autocrine growth factor that is overexpressed in human breast cancer cell lines. We have examined immunohistochemical PCDGF expression in 206 paraffin-embedded human breast lesions and investigated its association with clinicopathological variables. PCDGF staining was observed in breast carcinoma, whereas it was almost always negative in benign breast epithelium. PCDGF expression was more common in invasive ductal carcinoma (80% cases positive) than in invasive lobular carcinoma (53% positive). PCDGF staining was almost never observed in lobular carcinoma in situ. Ductal carcinoma in situ expressed PCDGF in 66% of the cases, and this expression correlated strongly with nuclear grade. Similar correlation was observed between PCDGF expression and histologic grade of invasive ductal carcinoma. Average Ki-67 index of PCDGF-negative/weakly positive invasive carcinomas (30.3) was significantly lower than that of strongly PCDGF-positive tumors (48.8, P=0.01). A larger percentage of tumors that expressed PCDGF with a staining intensity of 2+ or 3+ were p53 positive (44%) than were PCDGF-negative tumors (25%), P=0.02. PCDGF expression was independent of c-erbB-2 overexpression and of ER and PR status. Our study provides the first evidence of high incidence of PCDGF expression in human breast cancer in which it correlates with clinicopathological variables such as tumor grade, proliferation index, and p53 expression. These characteristics, as well as the virtual absence of expression in benign breast tissue, suggest an important role of PCDGF in breast cancer pathogenesis and make it a potential novel target for the treatment of breast cancer.