Immunohistochemical Staining For Bcl-2 Research Articles

A novel triad for the diagnosis of endometriosis, the short anogenital distance combines with high endometrial BCL2 and low endometrial FASL.

To investigate the anogenital distance from the upper verge of the anus to the posterior fourchette (AGDAF), FASL, and BCL2 combination as a reliable and non-invasive tool for the diagnosis of endometriosis. This study included 100 women with endometriosis and 50 women without endometriosis as the control group. All cases underwent history taking, body mass index (BMI) measurement, AGD measurement, and FASL and BCL2 immunohistochemical staining of the eutopic endometrial tissue. This study included 150 women divided into endometriosis and control groups. Endometriosis cases significantly had shorter AGDAF, 22.9 ± 2.6 mm, compared with the control group, 27.3 ± 3.5 mm (P < 0.001). Lower FASL and higher BCL2 expression were associated with endometriosis (P < 0.001). The combined measurement of AGDAF (cut-off point 24.55 mm) with FASL and BCL2 was associated with endometriosis (P < 0.001). The combined diagnostic sensitivity, specificity, positive predictive value, and negative predictive value of AGDAF, FASL, and BCL2 were 83%, 78%, 87.3%, and 69.6%, respectively. The area under the curve was greater for AGDAF, FASL, and BCL2 in combination than for individual measurements. Combining short AGDAF with high BCL2 and low FASL is a highly sensitive, non-invasive diagnostic tool for endometriosis.

Improving fine needle aspiration to predict the tumor biological aggressiveness in pancreatic neuroendocrine tumors using Ki-67 proliferation index, phosphorylated histone H3 (PHH3), and BCL-2

IntroductionSurgery is the only known cure for sporadic pancreatic neuroendocrine tumors (PNETs). Therefore, the prediction of the PNETs biological aggressiveness evaluated on endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) has a significant impact on clinical management. The proliferation rate of Ki-67 in PNETs can help to predict the biological aggressiveness of the tumor. In addition, there is a relatively new proliferation marker called phosphorylated histone H3 (PHH3) that can identify and quantify dividing cells in tissue samples, which is a marker highly specific to mitotic figures. Other markers such as BCL-2 also contribute to tumorigenesis and may be involved in the differentiation of neuroendocrine cells. Materials and methodsA retrospective observational study was performed on patients undergoing surveillance for PNETs from January 2010 to May 2021. Data collection included the patients' age, sex, tumor location, tumor size in the surgical specimen, and tumor grade in FNA. The 2019 World Health Organization (WHO) classification guideline was followed to diagnose PNETs, including grade and stage. Immunohistochemical stainings for Ki-67, PHH3 and BCL-2 in PNETs were performed. ResultsAfter excluding cell blocks containing fewer than 100 tumor cells, 44 patients with EUS-FNA and surgical resection specimens were included in this study. There were 19 cases of G1 PNETs, 20 cases of G2 PNETs, and 5 cases of G3 PNETs. The grade assigned based on the Ki-67 index was higher and more sensitive than that based on the mitotic count using H&E slides in some cases of G2 and G3 PNETs. However, there was no significant difference between the mitotic count using PHH3-positive tumor cells and the Ki-67 index to grade PNETs.All grade 1 tumors (19 cases) on surgical resection specimens were correctly graded on FNA (100 % concordance rate). Within the 20 G2 PNETs, 15 cases of grade 2 on surgical resection specimens were graded correctly on FNA based on the Ki-67 index only. Five cases of grade 2 PNETs on surgical resection specimens were graded as grade 1 on FNA when using only the Ki-67 index. Three of five grade 3 tumors on surgical resection specimens were graded as grade 2 on FNA based on the Ki-67 index only. Using only FNA Ki-67 to predict PNET tumor grade, the concordance (accuracy) rate was 81.8 % in total. However, all these eight cases (5 cases of G2 PNETs and 3 cases of G3 PNETs) were graded correctly by using the Ki-67 index plus mitotic rate (using PHH3 IHC stains).Four of 18 (22.2 %) patients with PNETs were positive for BCL-2 stain. In these 4 cases positive for BCL-2 stains, 3 cases were G2 PNETs and one case was G3 PNETs. ConclusionGrade and the proliferative rate in EUS-FNA can be used to predict the tumor grade in surgical resection specimens. However, when using only FNA Ki-67 to predict PNET tumor grade, about 18 % of cases were downgraded by one level. To solve the problem, immunohistochemical staining for BCL-2 and especially PHH3 would be helpful. Our results demonstrated that the mitotic count using PHH3 IHC stains not only improved the accuracy and precision of PNET grading in the surgical resection specimens, but also could reliably be used in routine scoring of mitotic figures of FNA specimens.

Construction of Fe-doped ZIF-8/DOX nanocomposites for ferroptosis strategy in the treatment of breast cancer.

Breast cancer has become one of the top five commonest causes of cancer death. The use of ferroptosis to induce the generation of reactive oxygen species (ROS) in cancer cells presents a promising and potential strategy for cancer treatment. Herein, a series of facile bimetallic nanoparticles (x% Fe-doped ZIF-8) were synthesized and tested, and doxorubicin (DOX), a classic drug for breast cancer therapy, was encapsulated. After comparing the ratios of Fe2+/(Fe2+ + Zn2+), 7% Fe-doped ZIF-8 (7FZ) was found to be the most suitable particle for medical application. The drug loading efficiency of DOX@7FZ was 58.01 ± 0.02%. The pH-sensitive DOX@7FZ was degraded and DOX was released in lysosomes once internalized. Both the intracellular content of iron and ROS increased significantly. Meanwhile, the cell viability declined to 13.98% in 24 h at a concentration of 60 μg mL-1 and the IC50 was 42.68 μg mL-1. Moreover, the expression of Bcl-2 and GPX-4 proteins decreased in a time-dependent manner, indicating that DOX@7FZ was able to enhance the ROS level in cancer cells via a synergistic effect between apoptosis and ferroptosis. The mechanism of action of DOX@7FZ was further verified using hematoxylin and eosin staining and immunohistochemical staining of Bcl-2 and GPX-4. These remarkable characteristics of DOX@7FZ may inspire further advancements in the treatment of breast cancer.

Identification of MYC/BCL2 Double-expressers Lymphomas in Diffuse Large B Cell Lymphoma Patients and Association with their Prognostic Parameters and Survival Rates: A Single Centre Experience

Introduction: Diffuse large B cell lymphoma (DLBCL) is a heterogenous disease some of which show co-expression of c-MYC and BCL2 and they are known as double expressers (DE) lymphoma. DE lymphoma has been shown to be associated with aggressive clinical course and poor outcome in comparison to those without co-expression. The association of DE lymphomas with their prognostic parameters and the overall survival rate of DE compared to non-DE lymphoma patients were investigated. Method: 66 formalin-fixed paraffin-embedded DLBCL cases were subjected to c-MYC and BCL2 immunohistochemical staining. Results: Out of 66 cases, 25(37.9%) cases showed c-MYC and BCL2 co-expression. Majority of the patients were in age category of ≥ 60 years (68.0%). Male and female distribution were almost equal. DE lymphoma was significantly associated with ABC subtype in comparison to GCB subtype (p= 0.026). Median survival time for DE was 29 months and non-DE was 34 months. Log rank analysis showed there was no significant difference in the overall survival rate of DE and non-DE patients (p=0.401). Conclusion: This study showed significant association between DE lymphomas and ABC subtype. Identification of DE lymphoma with ABC subtype may guide the clinicians in identifying patients with poorer outcome hence a more aggressive treatment may be offered to this group.

Deleterious effect of Toluene on Rat cerebrum cortex and The exert effect of resveratrol: (Histological, Immunohistochemical and Ultrastructural study)

Abstract: Background: Toluene is a chemical hydrocarbon that affects mainly the central nervous system. Resveratrol has a large spectrum of beneficial health effect. Aim: The study aimed to assess the protective effect of Resveratrol (RES) on cerebrum injury in rat caused by toluene (TN). Materials & Methods: Thirty adult male Wistar albino rats were used in this study, each weighting 200-250 g, aging 4-6 months. They were separated into five equal groups, Control group: The animals had only regular diet and tap water, Corn oil group: was given 0.1 ml/10 g/day corn oil, intraperitoneally, TN-treated group: The animals received Toluene intraperitoneally with a dosage of 500 mg/kg/day (n ¼ 7) (0.1 g/ml, in corn oil) (0.5ml/kg/day) for 14 days, RES group: The animals received resveratrol 10 mg/kg/day intraperitoneally for 14 days, and TN+RES treated group: The animals received toluene alongside supplemented with resveratrol (as previous doses) for 14 days. All rats were anesthetized, scarified and the brain was dissected out. Parts of brain tissues were processed for tissue homogenates for measuring MDA, SOD, Gpx. And other parts of the brain tissues were subjected to histopathological examination (light and electron microscopic examination). Immunohistochemical staining for BCL2, BAX, GFAP and synaptophysin were performed with morphometric and statistical analysis. Results: Toluene revealed increment in MDA together with a decrement of SOD and Gpx activity. Moreover, Toluene exhibited obvious histopathological deterioration with over expression of Bax protein and glial fibrillary acidic protein (GFAP) and down expression of Bcl2and synaptophysin. RES restored the biochemical profile of oxidative damage and improved the histopathological and immunohistochemical picture. Conclusion: In conclusion, our results speckled a bright on the probable effective role of RES on cerebrum injury induced by toluene and provide available suggestion on the potential clinical effectiveness of RES supplementation in case of cerebrum injury.

Abstract C098: Racial disparity in the prevalence of BCL-2 expression and associations with breast cancer survival in the Breast Cancer Care in Chicago study

Abstract Background: BCL-2 is an antiapoptotic protein that regulates apoptosis and has been associated with poor prognosis for many cancer sites. Despite a function that encourages tumorigenesis, increased BCL-2 expression has been associated with improved clinical outcomes in ER+ breast cancers, with mixed results observed for ER/PR- cancers. BCL-2 has been hypothesized to be an indicator of intact and fully functioning hormone receptor pathways and has also been hypothesized to facilitate cell death after treatment through mitochondrial priming. BCL-2 expression correlates strongly with hormone receptor expression; therefore, its association with better prognosis may be due to the strong effect ER/PR status has on survival, or due to its own biologic mechanism. To date, no studies have been conducted on racial differences in BCL-2 expression. We characterized racial/ethnic differences in BCL-2 expression and the association of BCL-2 expression with breast cancer specific survival among participants in the Breast Cancer Care in Chicago Study (BCCC). Methods: The BCCC was a cross-sectional study of 989 recently diagnosed non-Latina (nL) White, nL Black and Latina breast cancer patients diagnosed with a first primary breast cancer aged 30-79 in Chicago from 2005-2008. Tumor tissue was available for BCL-2 immunohistochemical staining for 264 patients. BCL-2 staining scores of 0 were classified as negative and all positive scores (1D, 1H, 2D, 2H, and 3) were classified as positive. Cox proportional hazards models were conducted to estimate the association of BCL-2 expression with breast cancer-specific death. Results: Roughly three-fourths of tumors (77%) stained positive for BCL. The prevalence of positive BCL staining varied by race/ethnicity (p=0.007), being highest for nL Whites (87%) and lowest for nL Blacks (68%); variability in BCL staining appeared to be limited to ER/PR-negative tumors (N=53), for which prevalence of positive BCL staining appeared to be much higher for nL whites than for minority patients (44% vs. 16%, p=0.054). BCL-2 expression was associated with a strong protective effect on BC survival in age and race-adjusted models (HR=0.40, 95% CI: 0.19, 0.84) and BCL-2 expression remained strongly positively associated with protection from breast cancer death with additional adjustment for ER/PR status (HR=0.33, 95% CI: 0.12, 0.89). Within subgroups defined by ER/PR status, BCL2 expression was qualitatively associated with better survival for both ER/PR positive and ER/PR negative subtypes, although sample size was insufficient to conduct a formal stratified analysis. Conclusions: BCL-2 appears to be an independent predictor of improved prognosis for breast cancer even after controlling for ER/PR status. The apparently higher prevalence of BCL2 expression for nL White vs. nL Black patients may provide etiologic clues regarding disparities in BC survival. Citation Format: Jibril M. Alim, Kent Hoskins, Abeer M. Mahmoud, Virgilia Macias, Andre Kadjascy-Balla, Garth H. Rauscher. Racial disparity in the prevalence of BCL-2 expression and associations with breast cancer survival in the Breast Cancer Care in Chicago study [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C098.

Clinicopathological features of in situ follicular neoplasm and relations with follicular lymphoma in Japan

This study aims to investigate the clinicopathological features of in situ follicular neoplasm (ISFN) in Japan. ISFN is a rare condition formerly considered as an early precursor of follicular lymphoma (FL). This is a first original report of ISFN from Asian country. We reviewed 19 biopsy samples of ISFN. ISFNs were categorized into two groups: (1) ISFN, consisting of ISFN with strong positivity for BCL-2 immunohistochemical staining (IHC), and obvious translocation of BCL-2; and (2) ISFN-like FL, featuring cases without obvious translocation but having morphological features and characteristic IHC findings of ISFN. As control, we adopted obvious FL. For some cases showing coexisting ISFN and FL lesions in the same lymph node, we could conduct further clonality analysis for each lesion. Nine of the 19 cases of ISFN coexisted with FL or had a history of overt B- or T-cell lymphoma including FL. Statistical comparison among ISFN-like FL and FL showed no significant differences in pathological features. Molecular analysis suggested that ISFN lesion and FL lesion in the same lymph node each have a different clonality. ISFN coexists or associates with other overt lymphomas frequently.

Expression of Pro-Apoptotic Bax and Anti-Apoptotic Bcl-2 Proteins in Hydatidiform Moles and Placentas With Hydropic Changes

Abstract- Morphologic examination still forms the main diagnostic tool in the differential diagnosis of molar placentas. However the criteria are subjective and show considerable inter-observer variability among pathologists. The aim of the present study was to investigate the role of Bcl-2 and Bax immunostaining in the differential diagnosis of molar placentas. Bax and Bcl-2 immunohistochemical staining were performed in 19 molars (8 partial and 11 complete hydatidiform mole) and 10 non-molar (hydropic abortion) formalin-fixed, paraffin-embedded tissue samples. Ploidy analysis using flow cytometry had confirmed diploidy in hydropic abortions and complete hydatidiform moles and triploidy in partial hydatidiform moles. Bcl-2 expression was observed only in syncytiotrophoblasts, No immunoreactivity was detected in Cytotrophoblasts, and stromal cells, the total score averages of Bcl-2 immunoexpression in partial hydatidiform moles and hydropic abortions were significantly higher than in complete hydatidiform moles, whereas no significant difference was observed between partial hydatidiform moles and hydropic abortions. Bax immunoreactivity was observed in cytotrophoblasts, stromal cells and occasionally in syncytiotrophoblasts. No statistically significant difference in Bax immunoexpression total score was observed among various groups. Based on the results of this study, Bcl-2 immunostaining offers a potential adjunctive diagnostic tool to distinguish complete hydatidiform mole from partial hydatidiform mole and hydropic abortion, but not partial hydatidiform mole from hydropic abortion, Bax immunostaining cannot be helpful in this regard.

Comparative Histological Study between the Effect of Calcitonin versus Hormonal Replacement Therapy on Cartilage of Knee Joint of Ovariectomized Albino Rat

Background: Osteoarthritis (OA) is the most common age-related joint degenerative disease, which is associated with post menopausal females. Aim of Work: To compare the effect of hormonal replacement therapy (HRT) (Oestrogen and Progesterone) versus calcitonin administration on articular cartilage of the knee joints in murine model of osteoarthritis induced by ovariectomy. Materials and Methods: Thirty six adult female rats were divided into five groups. Group I: control; Group II: sham operated; Group III: ovariectomized rats: Group IV: ovariectomized then received HRT )1ml/kg(0.2ml)/day IM ((for 8 weeks); GroupV: ovariectomized then received calcitonin )2 IU/kg/day (0.04ml) SC( (for 8 weeks). The rats of each group were sacrificed after 8 weeks. Articular cartilage of knee joints were processed and stained with Haematoxylin and Eosin, Alcian blue, PAS, immunohistochemical staining of Bcl-2 and statistical analysis were applied. Results: Ovariectomy resulted in degeneration, apoptosis and deterioration of normal structure of articular cartilage, with decreased PAS reaction, decreased Alcian blue staining, and weak immunoreactivity for Bcl-2 when compared with the control and sham operated groups. Calcitonin and HRT supplementation after ovariectomy mostly prevented these changes. Conclusion: It could be concluded that OVX causes severe changes in articular cartilage and these changes are more evident with time. HRT had a significant effect in prevention of cartilage degradation in OVX model of OA. Calcitonin administration greatly improves the histological architecture of the articular cartilage. So calcitonin had a significant effect in prevention of cartilage degradation in OVX model of OA. It can be considered as a potential therapeutic agent in OA.

Increased glomerular Bax/Bcl2 ratio is positively correlated with glomerular sclerosis in lupus nephritis

IntroductionThe role of intrinsic pathway of apoptosis in pathogenesis of lupus nephritis (LN) is still not clear. We investigated the relation between the expression of two major proteins of the intrinsic pathway of apoptosis; bcl2 as an antiapoptotic protein and bax as a proapoptotic one; in renal tissue of LN. MethodsThe study included fifty paraffin embedded renal tissue obtained from renal biopsy specimens of LN patients (8 cases class II, 10 cases class III, 21 cases class IV and 11 cases class V) and five paraffin embedded apparently normal renal tissue obtained from nephrectomy specimens due to renal neoplasms as a control group. Immunohistochemical staining for bcl2 and bax antibodies was done. Ki67 immunohistochemical staining was done for class III and IV to assess the degree of proliferation. The number of intraglomerular bcl2, bax and ki67 positive cells per glomerular cross section was evaluated for each case. The results were analysed in different LN classes and correlated to different glomerular lesions. Results: The expression of bax and bcl2 proteins was higher in LN glomeruli compared to normal. The expression of bcl2 was significantly higher in class IV and was correlated to the degree of endocapillary hypercellularity. The bax to bcl2 ratio was significantly correlated to the percentage and degree of glomerular sclerosis. ConclusionThe intrinsic pathway of apoptosis interfere in the pathogenesis of lupus glomerulonephritis. The balance between bax and bcl2 proteins might have a role in regulating the progression of glomeruli from proliferative to sclerotic state.

Evaluation of the Prognostic Importance of c-Myc and Bcl-2 Expressions and the Presence of Epstein-Barr Virus in Classical Hodgkin Lymphoma.

Although classical Hodgkin lymphoma (cHL) has a relatively good prognosis, it also entails different treatment responses and involves patients who have different clinical courses. Our aim was to investigate c-Myc, Bcl-2 and EBV biomarkers in cHL and their relationship with the IPS score. c-Myc and Bcl-2 immunohistochemical staining with EBER in situ hybridization (EBER-ISH) was applied to the paraffin sections of 94 cases diagnosed as cHL. These cases were classified into two groups as low and high clinical symptoms according to the International Prognostic Scores (IPS). Positive results were obtained in 83 (88.3%) cases with c-Myc and 39 (43.5%) cases with Bcl-2 while EBER-ISH was found positive in 42 (44.7%) cases. No difference was found between the groups of low/high IP scores with respect to the positive or negative results of EBER-ISH, Bcl-2 and c-Myc. When Bcl-2 and c-Myc positive cases were grouped together and compared to the IP scores of the remaining cHL cases, again no difference was seen. Extranodal involvement and bone marrow involvement was observed in 25 (26.5%) and 9 (9.5%) cases, respectively. Similarly, no statistically significant differences was found between these groups according to their positivity with EBER-ISH, Bcl-2 and c-Myc. We could not find any relationship between Bcl-2, c-Myc and EBER-ISH positivity and the low/high IPS groups in cHL. New studies with larger series are needed in which more precise cut-off values are used and clinically and biologically heterogeneous groups of cHL patients are determined more clearly.

The Relationship Between Apoptotic Activity and Prognostic Factors in Neuroblastomas.

Prognostic parameters in determining risk groups for treatment in neuroblastoma are cellular differentiation, mitosis karyorrhexis index (MKI), N-myc amplification and age. However, additional prognosticators are still needed because patients can show unpredictable biological behavior. We aimed to study the prognostic significance of apoptotic activity in neuroblastomas. Thirty-five primary neuroblastoma were evaluated. The data including stage, treatment protocol, metastatic disease, survival, N-myc status, age and prognostic categorization were obtained from the clinical records. The differentiation and MKI were evaluated in hematoxylin and eosin stained slides. Apoptotic activity was assessed by both bcl-2 immunohistochemical staining and the transferase-mediated d-UTP-biotin nick end labeling (TUNEL) method. Data were correlated with established prognostic factors and clinical outcome. Twenty-five (71.4%) cases were located in the adrenal. Sixteen cases showed low and 19 high MKI. Thirty-three (%94) were immunopositive for bcl-2. TUNEL staining was negative in 2 cases. Of the remaining positive 33 cases, 14 had an apoptotic index of ≤2%, 11 of 2-4% and 8 of ≥4%. Cases located in the adrenal showed higher scores of bcl-2 positivity than extra-adrenal tumors. There was no statistical significance for both bcl-2 staining and apoptotic index in correlation with cellular differentiation, MKI, N-myc amplification, age and other clinical parameters. Statistical significance was observed between bcl-2 scoring and tumor localization. According to our results, apoptotic activity is unlikely to be a prognostic parameter in neuroblastomas. Some studies showing significant correlations between clinical outcome and both bcl-2 immunoscoring and apoptotic index, as assessed by the TUNEL method, differences in case numbers and methodology can explain these conflicting results. Larger series and different methodologies are needed to evaluate the prognostic value of apoptotic activity in neuroblastomas.

Evaluation of the Pathogenesis of Tumor Development from Endometriosis by Estrogen Receptor, P53 and Bcl-2 Immunohistochemical Staining

Objective:Endometriosis, one of the most common estrogen dependent gynecological disorders, can present as both benign and malignant disease. The prevalence of tumoral transformation is 0.7-1.6% and the most common tumors are clear cell and endometrioid carcinomas. Unfortunately, the pathogenesis of transformation is unknown. For this purpose, we examined molecular alterations in ovarian endometriosis and endometriosis-associated tumors.Methods:Using the data bank of Alzahra hospital pathology department and paraffin blocks from appropriate cases were identified. Sections were cut and stained for 3 markers: estrogen receptor, P53 and bcl2. Correlations between findings were investigated.Results:Nineteen cases of endometriosis-associated tumor and 19 cases of endometriosis were identified. Staining for bcl2 was documented in 14 of 19 (73.7%) of endometriosis-associated tumor cases and also 7 of 19 (36.8%) endometriosis cases (P=0.02). Only 3 of the 19 (15.8%) endometriosis-associated tumors exhibited positive staining for estrogen receptors, compared with 14 of 19 (73.7%) endometriosis cases (P<0.001). Positive staining for P53 was noted in 5 of 19 (31.6%) endometriosis-associated ovarian tumor samples but was absent in endometriosis samples (0%), (P =0.008).Conclusions:Endometriosis-associated tumors appear to be associated with overexpression of bcl2 and P53 and reduced expression of Estrogen receptor. These finding may help to diagnose tumoral transformation with a background of endometriosis.

Partial lack of BCL2 in follicular lymphoma: An unusual immunohistochemical staining pattern explained by ongoing BCL2 mutation

Follicular lymphomas are characterized by overexpression of BCL2 which, in the large majority of cases, is due to a t(14;18) translocation which juxtaposes the BCL2 locus to the immunoglobulin heavy chain locus (IGH). Here, we report partial absence of BCL2 immunohistochemical staining in a case of FL, due to a mutation in the part of BCL2 that encodes the epitope for the most frequently used antibody against BCL2. This finding shows that mutations in BCL2 occur in an ongoing process in follicular which can give rise to unusual immunohistochemical staining patterns.

Immunohistochemical biomarkers and FDG uptake on PET/CT in head and neck squamous cell carcinoma

ABSTRACTBackground. There is an exciting complementarity between the spatial resolution provided by molecular imaging of a single, often unspecific, biomarker on one hand and the more detailed biological profile achievable from a diagnostic biopsy using a panel of immunohistochemical (IHC) markers on the other. A number of previous studies have shown a relationship between glucose transport protein expression and 18F-Fludeoxyglucose (FDG) PET uptake. Here, FDG uptake is analyzed in relation to expression of a selected panel of IHC cancer biomarkers in head and neck squamous cell carcinomas (HNSCC).Material and methods. IHC staining for Bcl-2, β-tubulin-1 and 2, p53, EGFR, Ki-67, glutathione-S-transferase-π and p16 was performed on formalin-fixed paraffin embedded diagnostic biopsies from 102 HNSCC cases treated at Rigshospitalet during 2005–2009. The proportion of positive cells was used for analyses, except p16, which was scored according to EORTC guidelines. In all cases, maximal FDG standardized uptake value (SUV) metrics were extracted for the primary tumor, TSUVmax. Univariate linear regression and multiple linear regression of TSUVmax versus IHC markers were performed.Results. In univariate analyses, TSUVmax showed negative associations with Bcl-2 (p = 0.002) and p16 (p = 0.005) indices and positive association with β-tubulin-1 index (p = 0.003). On multivariate analysis, TSUVmax remained associated with β-tubulin-1 (p = 0.009), Bcl-2 (p = 0.03) and p16 (p = 0.03). All correlations had r-squared < 0.3.Conclusion. Statistically significant correlations were observed between the expression of IHC biomarkers and maximum FDG uptake in the primary tumor.

Prognostic value of Bcl-2 expression in squamous cell carcinoma of the larynx: a systematic review.

The aim of this systematic review was to determine the prognostic value of Bcl-2 immunostaining in patients affected by laryngeal squamous cell carcinoma. An appropriate search was conducted on PubMed to retrieve articles dealing with this topic. A double cross-check was performed on citations and full-text articles by 2 investigators independently to review all manuscripts and perform a comprehensive quality assessment. Of 115 abstracts identified, 15 articles were included. These studies reported on 1,150 patients with histologically confirmed diagnosis of laryngeal squamous cell carcinoma. Only a few studies showed a statistical correlation between Bcl-2 immunohistochemical expression and at least 1 of the clinical and histopathological parameters considered by the authors. Moreover, these findings were also discordant between them. Overall the studies analyzed suggested that Bcl-2 expression was statistically connected with N stage (2/14), grading (2/14), disease-free survival (3/14) and overall survival (5/14). Interestingly, all of the 3 studies investigating the relation between Bcl-2 and radioresistance showed significant results in terms of recurrence-free survival and overall survival. Our review strongly suggests that the immunohistochemical staining of Bcl-2 does not correlate with tumoral aggressiveness and prognosis of patients affected by laryngeal squamous cell carcinoma and treated with primary surgery. However, an interesting connection of this protein could be demonstrated with tumoral radioresistance. Further, high-quality prospective studies should be carried out to confirm this hypothesis.

Immunohistochemical expression of Bcl-2 and Ki-67 in oral lichen planus and leukoplakia with different degrees of dysplasia.

Oral lichen planus (OLP) is a chronic inflammatory disease of unknown cause. Malignant transformation in OLP lesions may be favored by changes in the expression of proteins that regulate cell proliferation and apoptosis. This study aimed to investigate these issues by immunohistochemical staining for Bcl-2 and Ki-67 and by correlating histopathological findings in samples from lesions of OLP and leukoplakia with epithelial dysplasia. Data for patients with OLP or leukoplakia with moderate or severe epithelial dysplasia recorded during 2006-2011 were retrospectively reviewed. The study samples represented 37 subjects with OLP (n=14), leukoplakia with moderate (n=8) or severe (n=6) epithelial dysplasia, and normal buccal mucosa (controls, n=9). New sections were subjected to histological examination and immunohistochemistry for Bcl-2 and Ki-67 in the basal layer, suprabasal layer, and inflammatory infiltrate, respectively. All basal layer sections stained either negative or positive in <10% of cells for Bcl-2 in OLP (92.9% and 7.1%, respectively) and control (77.8% and 22.2%, respectively) samples. In leukoplakia, 85.7% of sections indicated positivity in <10% of cells, and 14.3% indicated positivity in 10-26% of cells. Most OLP (42.9%) and leukoplakia (64.3%) sections stained positive for Ki-67 in >50% of cells. All suprabasal sections stained either negative or positive in <10% of cells for Bcl-2 in OLP (92.9% and 7.1%, respectively), leukoplakia (42.9% and 57.1%, respectively), and control (88.9% and 11.1%, respectively) samples. Suprabasal staining for Ki-67 was negative or positive in <10% of cells in OLP (14.3% and 85.7%, respectively), leukoplakia (7.1% and 92.9%, respectively), and controls (88.9% and 11.1%, respectively). Staining for Bcl-2 in inflammatory infiltrate in OLP was positive in 92.9% of sections. Expression of Bcl-2 may play a dual role in tumor development and progression. Increased cell proliferation in the epithelium may present a predisposition to cancer in OLP. The expression of Ki-67 can be considered as an adjunct marker for proliferative activity in lesions with malignant potential. The prognostic value of these immunomarkers in the evaluation of precancerous oral lesions requires further investigation.

Cyclooxygenase-2 and B-cell lymphoma-2 expression in cystitis glandularis and primary vesicle adenocarcinoma

BackgroundAlthough cystitis glandularis (CG) is a common benign urinary bladder epithelial abnormality, it remains unclear whether CG is a premalignant lesion. Cyclooxygenase-2 (COX-2) and B-cell lymphoma-2 (Bcl-2) overexpression has recently been reported as a potential tumor initiator or promoter. We evaluated and compared COX-2 and Bcl-2 expression in CG, chronic cystitis (CC), and primary vesicle adenocarcinoma (ADC) tissues.MethodsWe conducted a retrospective study to investigate COX-2 and Bcl-2 levels in CG and ADC. We obtained tissue samples from 75 patients (including 11 cases of CC, 30 typical cases of CG (CGTP), 30 cases of intestinal CG (CGIT), and 4 cases of ADC) between 1989 and 2009 from the Surgical Pathology Archives of the No. 2 People’s Hospital of Zhenjiang, affiliated with Jiangsu University. COX-2 and Bcl-2 immunohistochemical staining was performed on all tissues. Nine normal bladder epithelial specimens were evaluated as control samples. Correlations between COX-2 and Bcl-2 expression in CG were also analyzed.ResultsCOX-2 and Bcl-2 expression was higher in the ADC group compared to other groups (p < 0.05). COX-2 and Bcl-2 levels were higher in the CGIT group compared to the CGTP group (p = 0.000 for both). The CGIT and CGTP groups both showed higher COX-2 expression compared to the CC group (p = 0.000 for both). There was no difference in Bcl-2 expression between the CGTP and CC groups (p = 0.452). Additionally, the difference in COX-2 and Bcl-2 expression between the control and CC groups was also insignificant (p = 0.668 and p = 0.097, respectively). Finally, we found that COX-2 and Bcl-2 levels were positively related (r = 0.648, p = 0.000).ConclusionCOX-2 and Bcl-2 overexpression in the CG group suggests that CG, particularly the intestinal type, may be a premalignant lesion that converts into a tumor in the presence of carcinogens.

Prognostic Value of Mitotic Index and Bcl2 Expression in Male Breast Cancer

The incidence of male breast cancer (MBC) is rising. Current treatment regimens for MBC are extrapolated from female breast cancer (FBC), based on the assumption that FBC prognostic features and therapeutic targets can be extrapolated to MBC. However, there is yet little evidence that prognostic features that have been developed and established in FBC are applicable to MBC as well. In a recent study on FBC, a combination of mitotic index and Bcl2 expression proved to be of strong prognostic value. Previous papers on Bcl2 expression in MBC were equivocal, and the prognostic value of Bcl2 combined with mitotic index has not been studied in MBC. The aim of the present study was therefore to investigate the prognostic value of Bcl2 in combination with mitotic index in MBC. Immunohistochemical staining for Bcl2 was performed on tissue microarrays of a total of 151 male breast cancer cases. Mitotic index was scored. The prognostic value of Bcl2 expression and Bcl2/mitotic index combinations was evaluated studying their correlations with clinicopathologic features and their prediction of survival. The vast majority of MBC (94%) showed Bcl2 expression, more frequently than previously described for FBC. Bcl2 expression had no significant associations with clinicopathologic features such as tumor size, mitotic count and grade. In univariate survival analysis, Bcl2 had no prognostic value, and showed no additional prognostic value to tumor size and histological grade in Cox regression. In addition, the Bcl2/mitotic index combination as opposed to FBC did not predict survival in MBC. In conclusion, Bcl2 expression is common in MBC, but is not associated with major clinicopathologic features and, in contrast to FBC, does not seem to have prognostic value, also when combined with mitotic index.

Histological and immunohistochemical study on the protective effects of cetrorelix against chemotherapy-induced ovarian damage

Introduction Chemotherapy exerts several adverse effects on the ovaries and can lead to ovarian atrophy and infertility. Aim of the study This study was carried out to determine the possible protective effect of cetrorelix (a gonadotropine-releasing hormone antagonist) against the ovarian damage caused by a chemotherapeutic drug (cyclophosphamide). Materials and methods Thirty adult female albino rats were used in this study. They were classified into three main groups. Group I served as the control group. In group II, rats were injected intraperitoneally with cyclophosphamide. In group III, rats were injected intraperitoneally with cetrorelix in combination with cyclophosphamide. After 20 days, the rats were sacrificed and ovaries were excised and processed. Sections were stained with H&E, Masson's trichrome and immunohistochemical stains for Bcl2 (antiapoptotic gene) and FasL (proapoptotic factor). Results Cyclophosphamide injection resulted in ovarian damage in the form of depletion of ovarian follicles, apoptosis of granulosa cells (lining the follicular cavity) and cortical fibrosis. Bcl2 immunostaining was significantly decreased and FasL immunostaining significantly increased in granulosa cells in the rats of group II. Pretreatment with cyclophosphamide (group III) reversed these changes to a huge extent. Conclusion Cetrorelix reduced chemotherapy-induced ovarian damage by suppressing apoptosis.