Bcl-2 Antigens Research Articles

Synovial sarcoma of the neck: A malignant mimic of a common benign cyst

Synovial sarcomas are rare malignant neoplasms arising from mesenchymal stem cells, comprising 8–10% of soft tissue sarcomas. One in three cases occur in those under 20 years old. The lower extremity and trunk are the most common sites, with less than 10% occurring within the head and neck region. They have a variety of clinical and radiological appearances, with occasional cystic degeneration. Here we present a case of a 9-year-old boy who presented with a presumed benign branchial cleft cyst seen on ultrasound and CT. A unilocular cyst was found adjacent to the lateral horn of the thyroid cartilage. When excised, the cyst was 4 cm with a variably thick wall of fleshy tissue containing straw-coloured fluid. Histologically, the cyst wall was composed of mitotically active spindle cells in fascicles separated by oedematous and collagenous stroma with occasional gland formation. Immunohistochemistry supported a diagnosis of synovial sarcoma, with positive epithelial membrane antigen (EMA), Bcl-2 and cytokeratin antigens, and was confirmed by fluorescence in situ hybridisation with a SS18 rearrangement. This case highlights the important consideration of malignant entities in common benign presentations.

Віддалені результати гемітиреоїдектомії та їх прогнозування у хворих на однобічний вузловий зоб на фоні автоімунного тиреоїдиту

Мета роботи: визначення об’єму хірургічного втручання при вузловому зобі на фоні автоімунного тиреоїдиту з урахуванням активності процесів апоптозу та проліферації, індексу проліферативної активності.Матеріали і методи. На пункційному матеріалі ЩЗ, отриманому від 80 хворих із гістологічно верифікованим діагнозом вузловий зоб на фоні автоімунного тиреоїдиту до хірургічного лікування та через 1,5 – 3 роки після нього, проведені імуногістохімічні дослідження з використанням моноклональних антитіл проти Ki-67, Fas, FasL, Bcl-2 та P53 антигенів. За віддаленими результатами виділили дві групи хворих. До першої групи, яка становить 53 пацієнти, включені випадки, в яких не було розладів функції тиреоїдного залишку і змін об’єму залишеної частки залози порівняно з показниками до операції. Другу групу склали 27 хворих із клінічними і лабораторними ознаками післяопераційного гіпотиреозу середнього і тяжкого ступенів тяжкості, збільшенням об’єму залишеної частки залози на фоні прогресуючої лімфоїдної інфільтрації. Пацієнти обох груп у післяопераційному період отримували патогенетично обґрунтовану медикаментозну терапію автоімунного тиреоїдиту.Результати досліджень та їх обговорення. Проведені дослідження дозволяють розмежувати вибір об’єму операції – гемітиреоїдектомія чи тиреоїдектомія залежно від функціональної спроможності ЩЗ та показників апоптозу і проліферації у неураженій вузлом частці залози. Вважаємо, що тиреоїдектомія показана при поєднанні таких показників досліджуваних тестів: об’єм неураженої вузлом частки ЩЗ більший за 10 см3, рівень ТТГ більше за 3,55 мОД/л, вільного Т4 менше за 12,91 пмоль/л, рівень АТПО більше за 80,25 ОД/мл та АТТГ більше за 89,34 ОД/мл з показниками лімфопроліферативної активності та апоптозу, зокрема рівнів ІПА Ki-67 вищими за 10,55 %, ІІРК FasL – за 51 %, ІІРК Bcl-2 – за 90 %, та ІІРК p53 нижче 50 % і ІІРК Fas нижче 43 %.

Прогностичні маркери проліферації та апоптозу в пацієнтів на вузлові форми зоба на фоні автоіму-нного тиреоїдиту

Мета роботи: вивчити процеси апоптозу та проліферації в пункційному матеріалі щитоподібної залози (ЩЗ) при вузловому зобі на фоні автоімунного тиреоїдиту ВЗАІТ з використанням імуногістохімічного методу дослідження, а також визначити індекс проліферативної активності.Матеріали і методи. На пункційному матеріалі ЩЗ, отриманому від 75 хворих із гістологічно верифікованим діагнозом ВЗАІТ, проведено імуногістохімічне дослідження шляхом використання моноклональних антитіл проти Fas, FasL, Bcl-2, P53 та Ki67 антигенів.Результати досліджень та їх обговорення. Результати показали ступінь проліферативної активності тканини ЩЗ при ВЗАІТ. Виявлено високу проліферативну активність лімфоїдної тканини, помірну проліферативну активність тиреоцитів у ділянці лімфоїдної інфільтрації і низьку – поза нею. Виражена експресія Fas та FasL на тиреоцитах, у ділянках лімфоїдної інфільтрації побічно свідчить про те, що при ВЗАІТ відбуваються імунологічно зумовлені процеси апоптозу тиреоцитів. Збільшення кількості імунореактивних клітин, що експресують Ki67 у ділянці лімфоїдної інфільтрації і деструкції тиреоцитів, свідчить про регенерацію фолікулярного епітелію як компенсаторно-пристосувальну реакцію органа. Виражена експресія bsl-2 у лімфоцитах запобігає вхід клітин в апоптоз і подовжує час виживання клітин. Спостерігалася висока експресія протеїну р53 в ядрах тиреоцитів та фолікулярних просвітах, яку можна пояснити мутаціями гена р53, що дозволяє клітинам знайти толерантність до апоптичної дії ефекторів імунної системи.

Simultaneous Detection of Tumor Cell Apoptosis Regulators Bcl-2 and Bax through a Dual-Signal-Marked Electrochemical Immunosensor.

B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) are often used to monitor the apoptosis of tumor cells and evaluate cancer drug effect. In this work, a novel sandwich-type dual-signal-marked electrochemical biosensor was fabricated for simultaneous detection of Bcl-2 and Bax proteins. Reduced graphene oxide (RGO) layers were used as substrate to immobilize Bcl-2 and Bax antibodies for further capturing target antigens. CdSeTe@CdS quantum dots (QDs) and Ag nanoclusters (NCs) with antibody modification and mesoporous silica amplification were used as signal probes, which were proportional to the amount of Bcl-2 and Bax antigens. Mesoporous SiO2 can provide a larger surface area, more effectively charged by ethylene imine polymer or poly(diallyldimethylammonium chloride) to adsorb more probes. The Bcl-2 and Bax proteins were determined indirectly by the detection of oxidation peak currents of Cd and Ag using anodic stripping voltammetry, showing a good linear relationship in the protein concentration range from 1 ng/mL to 250 ng/mL. The detection limit of trace protein level was ∼0.5 fmol. The biosensor was further introduced to investigate Bcl-2 and Bax expressions from nilotinib-treated chronic myeloid leukemia K562 cells. With the increase of drug dosage and incubation time, the up-regulation for Bax and down-regulation for Bcl-2 were observed, which indicated that the apoptosis level of K562 cells could be regulated by Bcl-2 family. The ratio of Bax/Bcl-2 was further calculated for evaluation of its drug effect and apoptosis level. The limited cell amount for detection reached less than 1 × 10(3) cells, much lower than traditional methods. Furthermore, completely independent detection step and stable acid solutions containing Ag(+) and Cd(2+) for long-time storage contribute to reducing the error from the sample differences and avoiding the potential errors from the photodegradation of fluorescent probes, enzymolysis of DNA, or inactivation of enzyme during an excess experimental period.

Immunohistochemical Profile and Prognostic Significance in Chinese Primary Central Nervous System Lymphoma: Analysis of 132 Cases

Primary central nervous system lymphoma (PCNSL) is aggressive and confined to the central nervous system, including the brain parenchyma, leptomeninges, spinal cord, eyes or cranial nervous. Morphologically, approximately 95% of these tumors are DLBCL according to the new World Health Organization (WHO) classification. However, PCNSL has treatment outcome distinct from those of systemic DLBCL, as well as dismal prognosis than systemic DLBCL. Our goal was to determine the immunohistochemical profile and prognostic significance for 132 Chinese PCNSL cases. The expression of CD20, CD10, BCL-6, MUM1, CD138, BCL-2, and Ki67 antigens were observed by immunohistochemical method. All cases expressed CD20. CD10, BCL-6, and MUM1 were positive in 15.2% (20/132), 86.4% (114/132), 90.2% (119/132). CD138 was negative in 100% (39/39). BCL-2 was positive in 89.3% (108/121). The Ki67 antigen, a proliferative index, ranging from 1% to 100% (median 85.3%) and 76.5% (101/132) PCNSLs showed Ki67 ≥ 90%. Among 132 cases, 25 (18.9%) were classified as germinal center B-cell-like (GCB); 107 (81.1%) were classified as activated B-cell-like (ABC). The Ki67 index in 25 GCB was similar to that in 107 ABC (p=0.663>0.05). No significant correlation was found between Ki67 index and BCL-2 (p=0.225>0.05). Significant positive correlation was found between Ki67 index and BCL-6 expression (p=.000<0.05). Among 132 cases, 43 had complete data of treatment that received chemotherapy regimens based on HD-MTX. GCB and ABC had similar OS (p=0.969) and PFS (p=0.070). These findings support that PCNSL predominantly express an ABC immunophenotype and express high Ki67 index, and suggest that the proliferative activity of GCB was similar to ABC and the expression of BCL-6 but not BCL-2 was positively correlated with the malignant degree of tumors.Table 1Clinical characteristics.CharacteristicsPatients, n (%)Age (years); n=132≥60 y, n=53; <60 y, n=79Median (range)57 (21-85)Gender; n=132Male69 (52.3)Female63 (47.7)ECOG; n=430-18 (18.6)2-435 (81.4)LDH; n=43Normal25 (58.1)Elevated18 (41.9)Numbers of lesions; n=132148 (36.4)>284 (63.6)Involvement of deep structures; n=132Absence43 (32.6)Presence89 (67.4)Table 2Hans classification.CD10BCL-6MUM1ImmunoprofilePCNSL, n (%)+++GCB16 (12.1)++-GCB3 (2.3)+-+GCB0 (0)+--GCB1 (0.7)-+-GCB5 (3.8)-++Non-GCB90 (68.2)--+Non-GCB12 (9.1)---Non-GCB5 (3.8)Table 3Chang classification.CD10BCL-6MUM1ImmunoprofilePCNSL, n (%)++-GCB (Pattern A)3 (2.4)+--GCB (Pattern A)1 (0.8)-+-GCB (Pattern A)5 (3.9)+++activated GCB (Pattern B)16 (12.6)+-+activated GCB (Pattern B)0 (0)-++activated GCB (Pattern B)90 (70.9)--+activated non-GCB (Pattern C)12 (9.4) [Display omitted] [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Diagnostic value of CD10 and Bcl2 expression in distinguishing cutaneous basal cell carcinoma from squamous cell carcinoma and seborrheic keratosis

The distinction between cutaneous basal cell carcinoma (BCC), squamous cell carcinoma (SCC) and seborrheic keratosis (SK), which are common entities in clinical practice, can be difficult clinically and histologically. CD10 and Bcl2 antigens are important factors in tumor growth, survival and spread. The aim of the present study is to define the frequency of CD10 and Bcl2 expression in such cutaneous tumors and its relation to the clinicopathological characteristics as well as their possible diagnostic utility. CD10 and Bcl2 immunohistochemistry was performed on 30 BCC, 20 SCC and 15 SK. 93.3% of SK cases and 53.3% of BCC cases showed significant expression of CD10 in tumor cells when compared either with each other or with SCC cases (100% negative). Stromal CD10 expression was positive in 50% of BCC cases and 75% of SCC cases. Stromal CD10 expression was significantly higher in high risk BCC and BCC with infiltrating deep margins; furthermore, it showed a significant positive correlation with grade of SCC. A significant inverse correlation between CD10 expression in stromal and tumor cells of BCC was present. Bcl2 was significantly expressed in 93.3% of SK cases and 80% of BCC cases when compared with SCC cases (100% negative). It was found that for distinguishing BCC from SK, only CD10 expression in tumor cells provided a high diagnostic value with positive likelihood ratio (PLR) was 7.00. In addition, CD10 and Bcl2 expression in tumor cells could give convincing diagnostic value to distinguish SCC from SK (PLR=15.00 for each marker). Moreover, for differentiating BCC from SCC, only Bcl2 in the tumor cells could provide a high diagnostic value (PLR=5.5). In conclusion, CD10 and Bcl2 can help in differentiating cutaneous BCC from SK and SCC. The overexpression of CD10 in the stromal cells of SCC and some variants of BCC suggests the invasive properties of such tumors.

The Immunoexpression of Cd34, Bcl-2, and Ki-67 Antigens in Sinonasal Inverted Papillomas

Numerous molecular markers of sinonasal inverted papillomas (IP) were investigated in the past; however, significance of angiogenesis and inhibition of apoptosis were not well documented. This study was designed to determine expression of angiogenic marker CD34 antigen, antiapoptotic marker Bcl-2 oncoprotein, and proliferative marker Ki-67 antigen in the group of patients with IP. We matched up these findings to the group of patients with sinonasal carcinoma (SNC) and used chronic rhinosinusitis (CRS) patients as control group. In addition, we compared expression of the markers among IP patients who displayed distinctly different patterns of clinical behavior. Tissue samples were obtained from 46 surgically treated patients; 18 of them had a diagnosis of IP, 9 had documented SNC, and the remaining 19 patients had CRS. All specimens were stained using immunohistochemistry techniques for CD34 (mean vessel density [MVD]), Bcl-2, and Ki-67 antigens. Morphometry was evaluated by computer image analysis system. We noted statistically significant differences in expression of CD34 antigen, Bcl-2 protein, and Ki-67 antigen (for all groups, ANOVA p < 0.001) among the investigated groups. The mean value of CD34 antigen was significantly higher in the IP group than in the CRS group, but it was below the levels of the SNC group. Compared with the cases not complicated by recurrence, the patients with recurrent IP exhibited higher MVD levels, while levels of bcl-2 and Ki67 protein expression did not differ in a significant way between recurrent and nonrecurrent cases. The significant positive correlations were observed between Bcl-2 protein and Ki-67 antigen in IP and SNC groups and between Bcl-2 protein and CD34 antigen in the CRS group. Our findings underscore importance of angiogenesis in the development and prognosis of IP and support further investigation of this aspect of IP tumor growth.

Bovine herpesvirus type 5 infection regulates Bax/BCL-2 ratio

Bovine herpesvirus 5 (BoHV-5) is an α-herpesvirus that causes neurological disease in young cattle and is also occasionally involved in reproductive disorders. Although there have been many studies of the apoptotic pathways induced by viruses belonging to the family Herpesviridae, there is little information about the intrinsic programmed cell death pathway in host-BoHV-5 interactions. We found that BoHV-5 is able to replicate in both mesenchymal and epithelial cell lines, provoking cytopathology that is characterized by cellular swelling and cell fusion. Viral antigens were detected in infected cells by immunofluorescence assay at 48 to 96 h post-infection (p.i.). At 48 to 72 h p.i., anti-apoptotic BCL-2 antigens were found at higher levels than Bax antigens; the latter is considered a pro-apoptotic protein. Infected cells had increased BCL-2 phenotype cells from 48 to 96 h p.i., based on flow cytometric analysis. At 48 to 96 h p.i., Bax mRNA was not expressed in any of the infected cell monolayers. In contrast, BCL-2 mRNA was found at high levels at all p.i. in both types of cells. BoHV-5 replication apparently modulates BCL-2 expression and gene transcription, enhancing production of virus progeny.

Analysis of the immunohistochemical subtypes and prognosis of primary diffuse large B cell lymphoma of the central nervous system

To analyze the subtypes of primary diffuse large B cell lymphoma of the central nervous system (CNS DLBCL) and to explore the relationship between the subtype classification and prognosis. Immunohistochemical staining was used to determine the expression of CD20, CD3, CD10, Bcl-6, Mum-1, CD5, Bcl-2, Ki-67, FOXP-1, GCET-1, BLIMP-1 and LMO-2 antigens on paraffin-embedded sections of 47 cases. Hans, Choi and Tally subtypes were classified, and univariate and multivariate analyses were used to elucidate the relationship between the subtypes and prognosis. In the 47 cases, the expression of Bcl-2 in the tumor cells was 46.8%, CD10 4.3%, Bcl-6 70.2%, Mum-1 53.2%, GCET-1 36.2%, BLIMP-1 4.3%, FOXP-1 63.8% and LMO-2 19.2%. The positive rate of Ki-67 was 30% to 95%, with a median of 80%, of which 12 cases (25.5%) was > or = 90%. The Hans subtype classification showed 16 cases (34.0%) were of GCB type and 31 cases (66.0%) of non-GCB type. The Choi subtype classification showed 16 cases (34.0%) were of GCB type and 31 cases (66.0%) of ABC type. The Tally subtype classification showed 6 cases (12.8%) were of GCB type and 41 cases (87.2%) of non-GCB type. The results of this study show that there is no significant correlation between the three subtypes and prognosis. The prognosis is correated with post-operative radiotherapy, chemotherapy and MTX therapy.

Immunohistochemical Detection of Factors Related to Cellular Proliferation and Apoptosis in Radicular and Dentigerous Cysts

Introduction This study proposed to investigate aspects of cell proliferation and death in the epithelium of radicular (RCs) and dentigerous (DCs) cysts. Methods Serial sections of 17 RCs and 9 DCs were prepared for immunohistochemical detection of caspase-3, Bcl-2, and Ki-67 antigens. Results Caspase-3 was detected mainly in the suprabasal and superficial epithelial cells of RCs and DCs, whereas Ki-67 was detected predominantly in the basal layer. Both markers had significant expression in hyperplastic epithelium related to an intense inflammation in the capsule. Immunoreactivity for Bcl-2 was restricted to the basal layer and was significantly higher in atrophic epithelium of DCs than that of RCs. Conclusions These results suggest that epithelial proliferation is balanced by apoptosis and that the presence of inflammation inhibits the Bcl-2 expression. DCs and RCs have different formation mechanisms but have similar biological behavior in the presence of intense inflammatory infiltrate.

Selected Immunohistochemical Prognostic Factors in Endometrial Cancer

The objectives of this study were to assess the immunohistochemical expression of p53, bcl-2, c-erbB-2, Ki-67, estrogen (ER) and progesterone (PR) receptors, matrix metalloproteinase-7 and -26 (MMP-7 and MMP-26) in endometrial cancer patients and to assess the relation between steroid receptor positivity and other markers. Experimental prospective study. Department of Obstetrics and Gynecology, Department of Genetics, Department of Pathology, Palacký University Medical School and University Hospital Olomouc. We studied 144 cases of primary untreated endometrial carcinoma in which the p53, bcl-2, c-erbB-2, Ki-67, ER, PR, MMP-7, and MMP-26 antigens were investigated with the use of immunohistochemical methods. We evaluated the correlations among immunohistochemical staining and the age, International Federation of Gynecology and Obstetrics stage, grading, depth of invasion, and metastatic spread to lymph nodes. Mean age was 65.7 years (range, 34-90 years). p53, bcl-2, c-erbB-2, Ki-67, ER, and PR were positive in 35 (24.3%), 100 (69.4%), 41 (28.4%), 65 (45.1%), 115 (79.8%), and 127 (88.1%) cases, respectively. Matrix metalloproteinases were evaluated in a group of 70 patients, wherein MMP-7 was positive in 33 patients (47.1%) and MMP-26 was positive in 40 patients (57.1%). The expression of MMP-7 decreased with higher patient age. p53 and Ki-67 overexpression was found to be related to poor differentiation. Immunostaining for bcl-2 correlated with the positivity of steroid receptors status, whereas immunostaining for c-erbB-2 correlated inversely with ER-positive group of cases. The overexpression of p53 and Ki-67 seems to indicate a more malignant phenotype, whereas bcl-2 expression in dependence of steroid receptor positivity could contribute to the identification of high-risk tumors.

Immunohistochemical analysis of the S100A1, S100B, CD44 and Bcl-2 antigens and the rate of cell proliferation assessed by Ki-67 antibody in benign and malignant melanocytic tumours

Malignant melanomas usually have an unfavourable prognosis and poor response to chemotherapy. Deregulation of cell proliferation, programmed cell death and intercellular interactions are among several important mechanisms that might lead to malignant transformation of melanocytes and melanoma progression. The S100A1, S100B, Bcl-2 and CD44 antigens have all been described as being involved in different processes of melanoma progression. The expression of these antigens, as well as the rate of cell proliferation, was analyzed retrospectively in melanocytic tumours from 126 patients (32 males and 94 females, age ranging from 11 to 91 years). The series included benign (45 intradermal, 27 compound and eight displastic naevi) and malignant (39 primary and 14 metastatic) melanocytic tumours. The proliferating rate assessed by Ki-67 staining was lower in naevi than in melanomas, with a correlation coefficient of r = 0814. There was no overlap for rate of proliferation between benign and malignant tumours. The expression of S100A1 was low in benign melanocytic tumours and increased in malignant melanomas (r = 0.61). In contrast, a higher percentage of S100B antigen-positive cells were observed in benign melanocytic lesions than in melanomas (Pearson correlation coefficient, 0.627). In addition, positive immunostaining for S100B antigen in malignant melanomas corresponded with the areas with increased proliferating rate. The expression of Bcl-2 was lower in melanomas than in benign melanocytic tumours (r = -0.53). Bcl-2-negative areas within melanomas had an increased proliferating rate. The expression of CD44 showed a large variation both in benign and malignant melanocytic tumours. CD44 antigen expression was higher in melanomas with known metastases than in those without metastases, but this difference was not statistically significant.

Vitamin A family compounds, estradiol, and docetaxel in proliferation, apoptosis and immunocytochemical profile of human ovary endometrioid cancer cell line CRL-11731.

Endometrioid carcinoma represents approximately 10% of cases of the malignant ovarian epithelial tumors. According to literature, the vitamin A (carotenoids and retinoids) plays an essential role in cell proliferation, differentiation and apoptosis in both normal and neoplastic ovarian tissues. Apart from that, the retinoids alter a cytotoxic effect of chemiotherapeutics, i.e. docetaxel, on ovarian cancer cell lines. Retinoids act on cancer cells throughout different mechanism than taxanes, so they may be the potential candidates for the new treatment strategies of ovarian cancer. The aim of the study was to determine the effects of vitamin A family compounds (retinol, beta-carotene, lycopene, all-trans -, 9-cis - and 13-cis retinoic acid) on the growth and proliferation of CRL-11731 endometrioid ovary cancer cell line and on docetaxel and estradiol activity in this culture. The assay was based on [3H] thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in CRL-11731 cells were also studied. Among vitamin A family compounds retinol and carotenoids, but not retinoids, inhibited the growth of cancer cells in dose dependent manner. Only the concentration of 100 muM of docetaxel inhibited incorporation [3H] thymidine into CRL-11731 cancer cells. Retinol (33.4%+/-8.5), carotenoids (beta-carotene 20 muM 4.7%+/-2.9, 50 muM 2.2%+/-0.9; lycopene 10 muM 7.6%+/-0.8, 20 muM 5.2%+/-2.5, 50 muM 2.9%+/-1.2), and 13-cis retinoic acid (19.7%+/-2.2) combined with docetaxel (100 muM) significantly decreased the percentage of proliferating cells (p<0.0001). The antiproliferative action of lycopene alone and in combination with docetaxel was also confirmed in immunohistochemical examination (decreased the percentage of PCNA and Ki67 positive cells). Also retinol (10 muM) and lycopene (20 and 50 muM) combined with estradiol (0.01 muM) statistically decreased the percentage of proliferating cells compared to the control (p<0.0001) and estradiol (p<0.01, p<0.0001) group (63.5%+/-14.8, 61.0%+/-20.6, 15.0%+/-5.5 respectively). In our experiments, the compounds tested induced an apoptotic effect. Docetaxel and estradiol increased the percentage of apoptotic cells (71% apoptotic cells after administration of 10 muM all-trans retinoic acid combined with 0.01 muM estradiol, p<0.0001). beta-carotene, lycopene and all-trans retinoic acid alone and in combination with docetaxel were found to influence the expression of bcl-2 and p53 antigen in the cells examined. The results of our study justified an important role of vitamin A in the pathophysiology of the ovarian endometrioid cancer.

Immunohistology of amicrobial pustulosis of the folds

Amicrobial pustulosis of the folds (APF), is a rare pustular eruption, predominantly involving the cutaneous folds, the external auditory canals and the scalp, occurring in patients who exhibit a wide spectrum of autoimmune abnormalities. There is a lack of data on the immunohistology of APF. We report a new case of APF associated with features of systemic lupus erythematosus (LE). Extensive immunohistological examinations of lesional and nonlesional skin were performed. The results of our immunohistological studies indicate that immunostaining with p53, Bcl-2, and CD8 antigens could be useful in differentiating APF from closely related inflammatory conditions such as psoriasis and cutaneous LE. However, studies on a larger sample size including controls are needed to substantiate our findings.

Diagnosis of primary esophageal synovial sarcoma by demonstration of t(X;18) translocation: a case report

Synovial sarcoma (SS) is an uncommon soft tissue tumor occurring mainly in the periarticular region of the extremities of young adults. In this report, we describe a very rare occurrence of primary SS of the esophagus in a 72-year-old woman. Histologically, the tumor demonstrated biphasic morphologic findings associated with poorly differentiated areas. Tumor cells expressed vimentin, epithelial (EMA, CK7, AE1/3), bcl-2 and neuroectodermal (CD56, CD57, CD99) antigens. Differential diagnose included esophageal sarcomatoid carcinoma. Cytogenetic analysis confirmed the diagnosis of SS by identifying t(X;18) translocation. The literature of this very uncommon entity of the esophagus is reviewed.

A uniform activated B-cell–like immunophenotype might explain the poor prognosis of primary central nervous system lymphomas: analysis of 83 cases

Most primary central nervous system lymphomas (PCNSLs) in immunocompetent patients are diffuse large B-cell lymphomas (DLBCLs), characterized by poor prognosis, compared with systemic forms. A germinal center B-cell-like (GCB) origin of PCNSL was hypothesized on the basis of BCL-6 expression and ongoing mutational activity. Our goal herein was to determine, for 83 PCNSLs, the percentages of GCB and activated B-cell-like (ABC) phenotypes and their prognostic significance. CD10, BCL-6, MUM1, BCL-2, and CD138 antigens were immunohistochemically labeled on paraffin-embedded sections; the first 4 were positive in 2.4%, 55.5%, 92.6%, and 55.5% of the tumors, respectively. None of the 56 tested samples expressed CD138. Among the 82 patients with complete information, 79 (96.3%) were classified as ABC; 42 (51.2%) expressed BCL-6+ MUM1+, suggesting an "activated GCB" origin; 33 (40.2%) were exclusively MUM1+, and the remaining 4 (4.9%) were negative for all markers tested. These findings provide new insights into interpreting the poor PCNSL prognostic, which may, in part, be due to biologic aggressiveness associated with its activated B-cell-like pattern. We postulate assigning PCNSL a histogenetic "time-slot," overlapping late GC and early post-GC, that could explain the predominant ABC phenotype observed.

Correlation of pre-treatment apoptotic marker expression with response to neo-adjuvant chemotherapy in locally advanced breast cancer

701 Background: Neoadjuvant chemotherapy is used as standard treatment for locally advanced breast cancer. The anticipated end point of cytotoxic drug therapy is the induction of cell death through apoptosis. Understanding the balance between pro- and anti-apoptotic factors within the cell may be relevant to both the prediction and monitoring of response to chemotherapy. Methods: A prospective study of 36 consecutive patients who presented with locally advanced breast cancer was performed. Serial MR scans of the breast were performed to assess the response to treatment and patients were divided into “responders” or “non-responders” using RECIST criteria. Immunohistochemical staining of formalin-fixed, paraffin-embedded pre-treatment core biopsy specimens was performed using antibodies against Bax, Bad, Bid, Bcl-2, Bcl-XL, and p53 antigens. Results: Using RECIST criteria, the group was divided in to 25 responders and 11 non-responders. The pro-apoptotic proteins Bid, Bax and Bad demonstrated loss of protein expression in 0%, 33% and 47% respectively of tumours. Anti-apoptotic proteins Bcl-XL and Bcl-2 demonstrated over expression in 8% and 83% of tumours respectively. P53 over expression was seen in 44% of tumours. Tumour grade and over-expression of Bcl-XL predicted response to chemotherapy (p=0.017 and 0.023 respectively). Multivariate hierarchical regression analysis (controlled for grade) showed that Bcl-XL and Bad were significant predictors of response to neo-adjuvant chemotherapy Conclusions: Although this is a pilot study involving a small number of patients, apoptotic markers may be useful in predicting response to chemotherapy for breast cancer patients No significant financial relationships to disclose.

Can transforming growth factor-beta1 and retinoids modify the activity of estradiol and antiestrogens in MCF-7 breast cancer cells?.

Retinoic acid and transforming growth factor-beta (TGF-beta) affect differentiation, proliferation and carcinogenesis of epithelial cells. The effect of both compounds on the proliferation of cells of the hormone sensitive human breast cancer cell line (ER+) MCF-7 was assessed in the presence of estradiol and tamoxifen. The assay was based on [3H]thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in MCF-7 cells were also determined. Exogenous TGF-beta1 added to the cell culture showed antiproliferative activity within the concentration range of 0.003-30 ng/ml. Irrespective of TGF-beta1 concentrations, a marked reduction in the stimulatory action of estradiol (10(-9) and 10(-8) M) was observed whereas in combination with tamoxifen (10(-7) and 10(-6) M) only 30 ng/ml TGF-beta1 caused a statistically significant reduction to approximately 30% of the proliferative cells. In further experiments we examined the effect of exposure of breast cancer cells to retinoids in combination with TGF-beta1. The incorporation of [3H]thymidine into MCF-7 cells was inhibited to 52 +/- 19% (control =100%) by 3 ng/ml TGF-beta1, and this dose was used throughout. It was found that addition of TGF-beta1 and isotretinoin to the culture did not decrease proliferation, while TGF-beta1 and tretinoin at low concentrations (3 x 10(-8) and 3 x 10(-7) M) reduced the percentage of proliferating cells by approximately 30% (67+/-8% and 67+/-5%, P<0.05 compared to values in the tretinoin group). Both retinoids also led to a statistically significant decrease in the stimulatory effect of 10(-9) M estradiol, attenuated by TGF-beta1. In addition, the retinoids in combination with TGF-beta1 and tamoxifen (10(-6) M) caused a further reduction in the percentage of proliferating cells. Immunocytochemical analysis showed that all the examined compounds gave a statistically significant reduction in the percentage of cells with a positive reaction to PCNA and Ki 67 antigen. TGF-beta1, isotretinoin and tretinoin added to the culture resulted in the lowest percentage of PCNA positive cells. However, the lowest fraction of Ki 67 positive cells was observed after addition of isotretinoin. The obtained results also confirm the fact that the well-known regulatory proteins Bcl-2 and p53 play an important role in the regulation of apoptosis in the MCF-7 cell line, with lowered Bcl-2 expression accompanying easier apoptotic induction. The majority of the examined compounds act via the p53 pathway although some bypass this important proapoptotic factor.

Can transforming growth factor-beta1 and retinoids modify the activity of estradiol and antiestrogens in MCF-7 breast cancer cells?.

Retinoic acid and transforming growth factor-beta (TGF-beta) affect differentiation, proliferation and carcinogenesis of epithelial cells. The effect of both compounds on the proliferation of cells of the hormone sensitive human breast cancer cell line (ER+) MCF-7 was assessed in the presence of estradiol and tamoxifen. The assay was based on [3H]thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in MCF-7 cells were also determined. Exogenous TGF-beta1 added to the cell culture showed antiproliferative activity within the concentration range of 0.003-30 ng/ml. Irrespective of TGF-beta1 concentrations, a marked reduction in the stimulatory action of estradiol (10(-9) and 10(-8) M) was observed whereas in combination with tamoxifen (10(-7) and 10(-6) M) only 30 ng/ml TGF-beta1 caused a statistically significant reduction to approximately 30% of the proliferative cells. In further experiments we examined the effect of exposure of breast cancer cells to retinoids in combination with TGF-beta1. The incorporation of [3H]thymidine into MCF-7 cells was inhibited to 52 +/- 19% (control =100%) by 3 ng/ml TGF-beta1, and this dose was used throughout. It was found that addition of TGF-beta1 and isotretinoin to the culture did not decrease proliferation, while TGF-beta1 and tretinoin at low concentrations (3 x 10(-8) and 3 x 10(-7) M) reduced the percentage of proliferating cells by approximately 30% (67+/-8% and 67+/-5%, P

Hormone receptors in non-malignant meningiomas correlate with apoptosis, cell proliferation and recurrence-free survival.

A retrospective immunohistochemical and statistical analysis of patients with non-malignant meningiomas was undertaken to determine the correlation of steroid hormone receptor status with apoptosis, tumour cell proliferation, clinicopathological characteristics and prediction of recurrence. Paraffin sections from 51 primary intracranial totally resected benign and atypical meningiomas were immunohistochemically evaluated for the expression of progesterone (PR), oestrogen (ER) and androgen (AR) receptors, apoptotic rate, Bcl-2, p53 and Ki67 antigens. In addition to the above parameters, the mitotic index and the patients' clinicopathological data were statistically correlated and entered in a recurrence-free survival analysis. A high level of apoptotic cell death was associated with loss of PR expression by logistic regression analysis (P = 0.016). An inverse correlation existed between the mitotic index and PR counts (P = 0.009), while high Ki67 values correlated with increased ARs (P = 0.041). Atypical meningiomas had a lower ER staining score (P = 0.036). Multivariate analysis indicated that the absence of PR and large tumour size were significant factors for shorter disease-free intervals. The results suggest that ER expression is lost or reduced in atypical meningiomas, whereas loss of PR expression is an indicator of increased apoptosis and early recurrence. PRs and ARs may also influence tumour cell proliferation.