Bcl-2 Negative Patients Research Articles

A Simplified IPI Including BCL2 Identifies IPI 3 Patients with Poor Prognosis - a GLA/ Dshnhl and Lysa Collaboration

Introduction: The original IPI remains valid to identify prognostic groups of DLBCL patients (pts) treated with R-CHOP (Ruppert et al. Blood 2020) and has been used to define eligibility for more than 20 studies randomizing pts between R-CHOP and R-CHOP + X, none of which except the Polarix study met its endpoint. One reason why some studies failed was the inclusion of IPI 2/3 pts equalizing the difference between standard and experimental arm although significant survival differences in pts with higher IPI were detected. Following the WHO-HAEM5 classification, determination of BCL2 and MYC rearrangements is mandatory to diagnose DLBCL, NOS and HGBL- MYC/ BCL2. As surrogate markers, we included immunohistochemistry (IHC) for MYC and BCL2 into the IPI model hoping for a wider spread of survival curves and better selection of pts suitable for future clinical trials. Methods: We analyzed 2765 pts treated on DSHNHL (German high-grade lymphoma study group) phase 2/3 studies, calculated the IPI, and added results for BCL2 and MYC expressionto individual IPI factors using published thresholds (50%; 70%) (Ziepert et al. Haematologica 2020). In univariate analyses, log-rank tests were used. Proportional hazard models for each of the biologic markers were separately adjusted for IPI factors. Using a variable selection algorithm the final model was built. Hazard ratios (HR) with p-values are presented. Results: 2765 DLBCL pts by reference pathology treated on 12 DSHNHL phase 2/ 3 studies were eligible. Median age at diagnosis was 57 years (range: 18-80 years), 43% of pts were female. Median observation time was 5 years. All pts had been treated with R-CHOP or variations. At diagnosis, 610 pts (22%) scored an IPI of 0, IPI was 1, 2, 3, 4, or 5 in 895 (32%), 546 (20%), 436 (16%), 212 (8%) and 64 (2%) pts with 3-year PFS- and OS-rates of 95%,87%,77%,66%,66%,47% and 98%,92%,83%,71%,73% and 54%, respectively. Multivariate analyses confirmed the significance of IPI factors (age, stage, performance status, and LDH) except the number of extranodal sites (EN) which did not significantly contribute to the model in this cohort (HR EFS,PFS,OS=1.0). Accordingly, EN>1 was deleted also resulting in better distinction of IPI groups (3-year PFS- and OS-rates: 95%,87%,74%,63%,51% and 98%,92%,80%,70%,57%%). In the final model, adjusted for all IPI factors except EN>1, BCL2 and MYC expression remained significant (HR PFS=2.3 and 2.0, p<0.001 and p=0.003). These results were validated and confirmed in 1176 DLBCL patients from 5 French (LYSA) studies. EN>1 but also MYC expression did not significantly contribute the final model in the validation set. Thus, MYC expression was deleted from the final model. Using the IPI deleting EN>1 but including BCL2 expression we observed 3-year PFS-rates of 74% vs. 83% in BCL2-positive vs. BCL2-negative patients with IPI 2 (p=0.136). In pts with IPI 3 the 3-year PFS rates were 53% vs. 80% (p=0.011) (see figure 1); in pts with IPI 4 the respective 3-year PFS rates were 49%% vs. 71%. This latter difference was not significant due to small pt numbers (p=0.381). Conclusions: Deletingthe number of extranodal sites and adding BCL2expression tothe original IPI factors as the only readily available biologic marker allows building an IPI model which effectively separates IPI 3 pts in 2 groups with significantly different prognosis. Restricting eligibility for future clinical trials to DLBCL pts with IPI 3 (and IPI 4) who are BCL2-pos. may help to avoid overtreatment of pts well treated with R-CHOP and at the same time better select pts representing a group with truly unmet medical need who are most likely to benefit from experimental therapies.

Evaluation of Hexokinase 2 and Serum LDH in B-cell Non-Hodgkin’s Lymphoma Patients

Background and Objective: Tumor cells produce energy by glycolysis and Lactate Dehydrogenase and Hexokinase are the key part of glycolysis, The present study aimed to evaluate the expression of Hexokinase 2 and serum LDH in B-cell Non-Hodgkin’s Lymphoma patients Method: A total of 84 B-cell Non-Hodgkin’s Lymphoma Patients were enrolled in the study. HK-2 Marker was studied by Immunohistochemistry and LDH analysis was done by Cobas 6000 analyser method and correlated with Clinico-Pathological Parameters and Diagnostic panel. RESULT:73% of B-cell NHL patients had Abnormal LDH. It was significantly correlated with Multiple Disease site (P=0.002). In correlation with Diagnostic Panel (MIb1, CD10, CD2, MUM1, BCL6, BCL2) a higher trend of Abnormal LDH was seen in BCL-6 positive and BCL-2 negative Patients. 64% of patients had HK-2 expression. In correlation with Clinical parameters, it was significantly correlated with >50 age group, while in correlation with pathological parameters A significant correlation observed with single disease site as compared to multiple disease site (P=0.004), In correlation with Diagnostic panel A higher trend of HK-2 Expression was observed in CD10 Negative and BCL-2 Negative Patients. Further Univariate survival analysis of LDH and HK-2 showed similar Overall survival in Abnormal and normal LDH level patients and HK-2 Positive and negative Patients respectively. Further Intercorrelation of both markers, showed almost similar expression of HK-2 between patients with normal LDH level and abnormal LDH level. CONCLUSION: Abnormal LDH level was closely associated with patients having disease at multiple sites, suggesting its activity to be associated with the advancement of the disease, HK-2 positivity was related with older age group, strengthens the findings of increased altered metabolic activity with older age, They are also associated with BCL-2 negativity that indicates that anti-apoptotic activity of BCL-2 might be responsible for that Though there was no significant correlation of LDH,, HK-2 with overall survival, majority of the patients who were dead had abnormal LDH and half of them showed positive HK-2 expression Briefly our data indicates that LDH and HK-2 can be useful parameter in defining clinical and prognostic aspects in NHL patient but needs further evaluation with larger groups of patients Keywords: NHL, Altered metabolism, Hexokinase, LDH

Prognostic influences of B-cell lymphoma 2-positive expression on late recurrence in breast cancer.

B-cell lymphoma 2 (BCL2) has an antiapoptotic role, however, has resulted in it being a powerful favorable prognostic factor in breast cancer. Several studies revealed BCL2 is strongly associated with a lower rate of early recurrence after initial treatment in breast cancer patients, but study of a prolonged effect after 5 years is lacking. We investigated BCL2 as a prognostic factor in breast cancer in comparison to early and late recurrence. We retrieved data from 2,198 patients with primary breast cancer who underwent surgical treatment and adjuvant treatment at the breast cancer center between 2005 and 2015. Each molecular subtype was classified, and Ki-67 and BCL2 were also assessed by immunohistochemistry. BCL2 and the association between molecular subtypes were assessed in early and late recurrences, respectively. Five-year postrecurrence survival and BCL2 were also assessed. The BCL2-positive group was associated with favorable clinicopathologic characteristics. The time to recurrence was significantly longer in the BCL2-positive group (P = 0.035). Late recurrence after 5 years was higher in the BCL2-positive group (P = 0.029). In multivariate survival analysis, tumor size and BCL2-positive expression were the only independent prognostic factors for late recurrence (P = 0.004). In the patients with recurrence, 5-year postrecurrence survival was significantly higher in the BCL2-positive group (P < 0.001). Our result showed that prognosis was better in BCL2-positive patients compared to BCL2-negative patients at late recurrence. We suggested that BCL2 expression could be used as a marker to help determine additional adjuvant therapy or extended hormone therapy in hormone-dependent breast cancer.

Is Bcl-2 a predictive marker of neoadjuvant chemotherapy response in patients with urothelial bladder cancer undergoing radical cystectomy?

Background: Response to neoadjuvant cisplatin treatment in bladder cancer has been linked to expression of Bcl-2 protein by cancer cells. The objective of this study was to test Bcl-2 as a predictive marker of neoadjuvant cisplatin chemotherapy response in a patient cohort from randomized cystectomy trials.Methods: Tumor samples were taken from 247 patients with T2–T4 bladder cancer enrolled in two randomized trials comparing cystectomy with or without neoadjuvant chemotherapy. Tissue microarrays from pre-intervention transurethral resection specimens were assessed for Bcl-2 protein status by immunohistochemistry. Extension of staining above 10% was regarded as positive. Downstaging and survival ratios in relation to Bcl-2 immunoreactivity and neoadjuvant chemotherapy utilization were calculated using the log rank test and multivariate Cox proportional hazards regression analyses.Results: Bcl-2 expression was positive in 38% and negative in 62% of the 236 evaluable patients. Bcl-2 negative patients receiving neoadjuvant chemotherapy had a significant increase in survival (p = 0.009), while Bcl-2 positive patients showed no difference (p = 0.4). However, the interaction variable between neoadjuvant chemotherapy and biomarker status was not significant (p = 0.38). When the prognostic value was assessed in the no-chemotherapy group, 5-year overall survival times were significantly better among Bcl-2 positive patients than among Bcl-2 negative patients (42 months vs 33 months, p = 0.04), but again Bcl-2 status did not remain independent when other factors were adjusted. Also, in a multivariate analysis with all patients, Bcl-2 was not significant.Conclusions: Bcl-2 status is not an independent predictor of neoadjuvant cisplatin chemotherapy response and is not prognostic in muscle-invasive bladder cancer.

Interim 18F-FDG PET/CT and BCL2 for predicting the prognosis of patients with diffuse large B-cell lymphoma in the rituximab era.

In this study, we investigated the prognostic role of interim fluorine-18-fluorodeoxyglucose PET/computed tomography (I-PET/CT) and BCL2 in diffuse large B-cell lymphoma (DLBCL) patients treated with rituximab-containing chemotherapy. A total of 106 patients with newly diagnosed DLBCL underwent PET/CT scans at baseline and a subsequent I-PET/CT after the fourth cycle of chemotherapy. I-PET/CT was analyzed using the Deauville five-point score. The prognostic significance of I-PET/CT and BCL2 was evaluated by predicting progression-free survival (PFS) and overall survival (OS). With a median follow-up of 25 months, the 2-year PFS and OS were 78 and 95% in the I-PET/CT-negative group and 33 and 57% in the I-PET/CT-positive group (P<0.001). Patients with BCL2 positivity showed a shorter PFS than the BCL2-negative patients (P=0.002), but not in OS (P=0.068). In multivariate analysis, I-PET/CT and BCL2 were independent variables in assessing the outcome of PFS. I-PET/CT was also an independent factor for OS. Irrespective of the results of BCL2, patients with I-PET/CT positivity had a significant inferior outcome than the I-PET/CT-negative patients. In the I-PET/CT-negative group, patients with BCL2 positivity had a significantly shorter PFS than the BCL2-negative patients (P=0.005). I-PET/CT and BCL2 were two significant independent indicators of outcome for patients with DLBCL in the rituximab era. I-PET/CT was much better than BCL2 for patient stratification. BCL2 showed its supplementary role for further patient stratification when I-PET/CT was negative.

Role of Bcl-2 Immunohistochemical Expression As An Independent Biological Prognostic Marker At Diagnosis of Classical Hodgkin's Lymphoma

Abstract 4859 BACKGROUND:Most patients with classical Hodgkin's Lymphoma (CHL) are cured with primary treatment. However, a proportion of them fail to first line treatment needing to be rescued with subsequent lines of chemotherapy and/or autologous or allogeneic stem cell transplantation (auto-SCT and allo-SCT, respectively). The identification of clinical and biological characteristics of these patients at diagnosis is still a challenge and most prognostic systems fail to identify a proportion of patients with worse prognosis. In this context, different groups are currently analyzing several biological markers as determinants of clinical outcome. It has been reported that Bcl-2 immunohistochemical expression in Hodgkinxs Reed Sternberg cells (HRSC) might confer a worse prognosis. OBJETIVE:To analyze clinical outcomes following 1st line chemotherapy according to Bcl-2 expression at diagnosis of CHL. PATIENTS AND METHODS:CHL patients, older than 16 years old, receiving at least 1 line of treatment, were retrospectively studied for Bcl-2 expression in diagnostic samples. For this purpose, tissue sections were immunostained and semiquantitatively assessed for this marker. Cumulative incidence (CI) of treatment failure, treatment failure free survival (TfFS) and overall survival (OS) were defined as primary outcomes. Treatment failure was considered when a different treatment regimen was set up due to relapse after CR or failing to achieve CR following 1st line. RESULTS:103 patients (55 Bcl-2 positive patients and 48 Bcl-2 negative patients) were analyzed. Main patient and clinical features are shown in Table 1. Both cohorts were well balanced for the main prognostic factors. At a median follow up of 36m (2-221), 34m (2-140) for Bcl-2 negative patients and 38m (4.5-221) for Bcl-2 positive patients, CI of 3 years treatment failure was 19% and 50% for the negative and positive cohorts, respectively (p=0.02). 3 years TfFS after diagnosis was 75% for Bcl-2 negative patients vs 47% for Bcl-2 positive patients (p=0.1). Within the cohort of Bcl-2 negative patients, 9/48 (19%) underwent auto-SCT as part of rescue treatment while 18/55 (33%) of Bcl-2 positive patients received an SCT (13 auto-SCT and 5 allo-SCT) and 3 of them, a second SCT (allo) for the treatment of post-auto-SCT relapse. 3 years OS was 84.5% for negative and 86% for positive patients (p=NS). [Display omitted] CONCLUSIONS:According to these preliminary results, Bcl-2 expression in HRSC at diagnosis may constitute an independent biological prognostic marker in CHL patients, seemingly associated with worse outcome and need of second line chemotherapy. More studies and a longer follow up is needed in order to confirm that aggressive treatment strategies such as SCT may overcome the negative impact in survival of Bcl-2 expression in this population. Disclosures:No relevant conflicts of interest to declare.

Mechanism and clinical significance of anti-apoptotic gene bcl-2 expression in diffuse large B-cell lymphoma

To evaluate the molecular mechanism and prognostication of bcl-2 protein expression in different subgroups of diffuse large B-cell lymphoma(DLBCL) in Guangxi Zhuang Autonomous Region, China. Immunohistochemical stains for CD10, bcl-6, MUM-1, bcl-2 and NF-κB were performed in 214 cases of DLBCL. The Hans immunologic classification was applied to classify DLBCL into GCB and non-GCB subgroups. Using a dual-probe fluorescence in-situ hybridization (FISH) assay, IgH/bcl-2 gene translocation and bcl-2 amplification were analyzed. In 214 cases of DLBCL, 30.8% (66/214) of cases were GCB and 69.2% (148/214) were non-GCB. Twenty-seven point three percent (18/66) of GCB subgroups and 59.5% (88/148) of non-GCB subgroups had bcl-2 protein expression, with a significant difference (P < 0.01). IgH/bcl-2 translocation was positive in 3.7% (8/214) of cases, even majority of them (6/8) was found in GCB subgroup, while represented only 9.1% of GCB case. There was a significant difference (P = 0.02) in bcl-2 gene amplification between GCB (27/66, 40.9%) and non-GCB subgroup (86/148, 58.1%). Among non-GCB cases, the expression of bcl-2 was correlated with that of NF-κB expression and bcl-2 gene amplification (r = 0.216 and 0.219, respectively, P < 0.05). No similar correlation was observed in GCB cases. The overall survival time of bcl-2-positive patients (31.4 ± 3.8) months was shorter than that of bcl-2-negative patients (40.2 ± 4.2) months. In conjunction with immunophenotypes and clinical stages, the bcl-2 positive patients had a 1.89 times higher risk than that of the bcl-2 negative patients. Majority of the cases were prognostically unfavorable non-GCB subgroups among DLBCL, which were characterized by high frequency of bcl-2 gene amplification and low frequency of IgH/bcl-2 translocation. The anti-apoptotic gene bcl-2 was frequently expressed in non-GCB subgroups and closely related to the gene amplification and NF-κB activation. bcl-2 positive patients had more short overall survival times, would face significant higher risk of death, these results suggested that bcl-2 could be a prognostic marker independent to clinical staging and immunophenotyping.

Bcl-2 as a Predictive Factor for Biochemical Recurrence after Radical Prostatectomy: An Interim Analysis

The objective of this study was to determine Bcl-2 expression in localized prostate cancer and its potential role as a predictive factor for biochemical recurrence (BCR). This study included 171 Korean patients with newly diagnosed adenocarcinoma of the prostate who underwent radical prostatectomy (RP) without neoadjuvant therapy at a single center between February 2005 and May 2009. RP specimens obtained from these patients were analyzed for the expression of Bcl-2 using tissue microarray. The values of Bcl-2 and other clinicopathologic factors were evaluated. Statistical analysis was performed with contingency table analysis, chi-square tests, and a Cox proportional hazard model. Bcl-2 expression was immunohistologically-confirmed in 42 patients (24.6%). Bcl-2 expression was not associated with conventional clinicopathologic factors. Bcl-2 negative patients had a significantly longer mean BCR-free survival than Bcl-2-positive patients (p=0.036). Among several variables, a high Gleason score in the RP specimen (≥8), extraprostatic extension, seminal vesicle invasion (SVI), lymphovascular invasion (LVI), and Bcl-2 expression were significant predictors of BCR based on univariate analysis. Multivariate Cox proportional hazards analysis revealed that BCR was significantly associated with a high prostate specific antigen level (p=0.047), SVI (p<0.001), a positive surgical margin (p=0.004) and Bcl-2 expression (p=0.012). Bcl-2 expression in RP specimens is associated with a significantly worse outcome, suggesting a potential clinical role for Bcl-2. Post-operative Bcl-2 could be a significant predictor of outcome after RP.

Bcl‐2 but not FOXP1, is an adverse risk factor in immunochemotherapy‐treated non‐germinal center diffuse large B‐cell lymphomas

Non-germinal center (non-GC) phenotype, high level expression of the transcription factor forkhead box protein P1 (FOXP1) and anti-apoptotic protein Bcl-2 have been identified as unfavorable prognostic factors for diffuse large B-cell lymphoma (DLBCL) patients treated with chemotherapy. Our aim was to re-evaluate the prognostic impact of these biologic factors on the survival of the patients treated with immunochemotherapy. Expression of Bcl-2 and FOXP1, and cell of origin based on the Hans algorithm were determined immunohistochemically from samples of 117 de novo DLBCL patients treated with R-CHOP and R-CHOEP regimens, and correlated with clinical data. Consistent with our previous studies, no significant difference in 2-yr survival rates between the GC- and non-GC phenotypes was found. Both FOXP1 and Bcl-2 expression were associated with the non-GC phenotype. For all patients, no prognostic impact of FOXP1 positivity on survival was observed. However, Bcl-2 negative patients had a better survival as compared to Bcl-2 positive patients [failure free survival (FFS) 97% vs. 71%, P = 0.001 and overall survival (OS) 97% vs. 82%, P = 0.034]. When Bcl-2 related survival was analyzed in the GC- and non-GC subgroups, a significant prognostic effect of Bcl-2 on FFS was seen only in the non-GC group of patients (positive 65% vs. negative 100%, P = 0.011). A trend for the difference in OS was also observed (positive 84% vs. negative 100%, P = 0.082). The data demonstrate that expression of Bcl-2 and FOXP1 is associated with the non-GC phenotype, but only Bcl-2 expression continues to be of prognostic significance in DLBCL patients treated with immunochemotherapy.

Aggressive lymphoma

Non-germinal center (non-GC) phenotype, high level expression of the transcription factor forkhead box protein P1 (FOXP1) and anti-apoptotic protein Bcl-2 have been identified as unfavorable prognostic factors for diffuse large B-cell lymphoma (DLBCL) patients treated with chemotherapy. Our aim was to re-evaluate the prognostic impact of these biologic factors on the survival of the patients treated with immunochemotherapy. Expression of Bcl-2 and FOXP1, and cell of origin based on the Hans algorithm were determined immunohistochemically from samples of 117 de novo DLBCL patients treated with R-CHOP and R-CHOEP regimens, and correlated with clinical data. Consistent with our previous studies, no significant difference in 2-yr survival rates between the GC- and non-GC phenotypes was found. Both FOXP1 and Bcl-2 expression were associated with the non-GC phenotype. For all patients, no prognostic impact of FOXP1 positivity on survival was observed. However, Bcl-2 negative patients had a better survival as compared to Bcl-2 positive patients [failure free survival (FFS) 97% vs. 71%, P = 0.001 and overall survival (OS) 97% vs. 82%, P = 0.034]. When Bcl-2 related survival was analyzed in the GC- and non-GC subgroups, a significant prognostic effect of Bcl-2 on FFS was seen only in the non-GC group of patients (positive 65% vs. negative 100%, P = 0.011). A trend for the difference in OS was also observed (positive 84% vs. negative 100%, P = 0.082). The data demonstrate that expression of Bcl-2 and FOXP1 is associated with the non-GC phenotype, but only Bcl-2 expression continues to be of prognostic significance in DLBCL patients treated with immunochemotherapy. (Less)

Immunohistochemical Analysis of Non-Small Cell Lung Cancer: Correlation with Clinical Parameters and Prognosis

Non-small cell lung cancers (NSCLC) vary in their biologic behavior. Recurrence and tumor-related mortality may be attributable to molecular abnormalities in primary tumors. This study evaluated such immunophenotypes with regard to cell cycle regulation and proliferation, apoptosis, and angiogenesis, to determine their significance for patient outcome. Core biopsies from 219 patients with NSCLC were assembled on tissue microarrays, and the expressions of p16, p21, p27, cyclin B1, cyclin E, Ki-67, caspase-3, survivin, bcl-2, VEGF, and endostatin were evaluated by immunohistochemistry. Despite previously described prognostic relevance of some of the investigated molecules, many of those markers were not directly associated with recurrence or survival. However, there was a trend for p16 immunoreactivity to be associated with a good prognosis (57% vs. 42% in 5-yr survival) (p=0.071). bcl-2 expression was strongly correlated with a better outcome (65% vs. 45% in 5-yr survival) (p=0.029), and the hazard of death for bcl-2 positive patients was 0.42 times of that for bcl-2 negative patients (p=0.047). A multivariate analysis with Cox proportional hazards model confirmed that the lymph node status (p=0.043) and stage (p=0.003) were other independent prognostic factors. Our results suggest that p16 and bcl-2 provide prognostic information independent of the TNM stage in NSCLC.

Does Rituximab Influence the Prognostic Effect of Germinal Centre Phenotype in Diffuse Large B-Cell Lymphoma?.

PURPOSE: Germinal centre (GC) and non-GC phenotypes are predictors of outcome in diffuse large B-cell lymphoma (DLBCL), and can be used to stratify chemotherapy-treated patients into low- and high-risk groups. Our aim was to determine how combination of rituximab with chemotherapy influences GC-associated clinical outcome.METHODS: 95 DLBCL patients treated with rituximab in combination with CHOP or CHOEP regimen (immunochemotherapy) were included in the study. The median follow-up time was 27 months. 107 patients previously treated with chemotherapy served as a historical control group. BCL-6, CD10, and MUM1 expression was analyzed immunohistochemically by means of identifying GC and non-GC phenotypes. In addition, BCL-2 expression was determined. Expression data was correlated with survival parameters.RESULTS: Consistent with previous studies, chemotherapy-treated patients with GC phenotype displayed a significantly better overall survival (OS) than the non-GC group (70% vs 47% p=0.012). In contrast, GC-phenotype did not predict outcome in immunochemotherapy-treated patients. The OS for GC-group was 82% compared with 80% for those with non-GC phenotype (p=ns). Conversely, the relapse free survival (RFS) for patients with GC and non-GC phenotypes were 74% and 65% (p=ns), respectively. When the patients were grouped according to BCL-2 expression, survival rates among the group of BCL-2 negative patients tended to be better than in BCL-2 positive patients (OS, 94% vs 77%, p=0.097; RFS, 94% vs 66%, p=0.029).CONCLUSIONS: Rituximab in combination with chemotherapy seems to interfere with the prognostic value of GC- and non-GC phenotypes in DLBCL. However, rituximab may have a positive impact on outcome among the patients with BCL-2 negativity.

Role of the molecular staging and response in the management of follicular lymphoma patients

Bcl-2/IgH rearrangement is the molecular hallmark of follicular lymphoma which is present in 70 – 90% of cases at diagnosis. The significance of the bcl-2 rearrangement at onset of disease and of its clearing after treatment (molecular response) is still controversial.The aims of the present analysis are: to evaluate the incidence of bcl-2 rearrangement in blood and marrow in a cohort of patients systematically investigated at diagnosis, to describe the correlation between bcl-2 and presenting features, to clarify the correlation of molecular response with outcome.Of 98 patients studied at initial staging for the presence of bcl-2 rearrangement, 64 (65%) showed bcl-2/IgH rearrangement in peripheral blood (PB) and/or bone marrow (BM) (58 at Major Breakpoint Region, MBR, and 6 at minor cluster region, mcr) while no bcl-2/IgH rearrangement was detected in the remaining 34 (35%) (germline status). No statistically significant differences were found between bcl-2 positive and bcl-2 negative cases as concerns presenting clinical features and response to first-line therapy. The median event-free survival, EFS, was not reached for the bcl-2 negative patients in PB and was 11 months for bcl-2 positive patients (statistically significant, P = 0.01) and, similarly, the median EFS was not reached for the bcl-2 negative patients in BM and was 11 months for bcl-2 positive patients (statistically significant, P = 0.04).Of the 64 bcl-2 positive cases, patients were analysed for molecular response (48 in BM and 40 in PB): 16 were molecular responders in BM and 20 were molecular responders in PB. The median EFS was 19 months for molecular responders in PB and 9 months for non-responders; 1-year-EFS was 68% (95% CI; 49 – 88), for responders in PB and 42% (95% CI; 22 – 61) for non-responders (P = 0.05). The median EFS was 11 months both for molecular responders and non-responders in BM; 1-year-EFS was 52% for responders in BM (CI; 30 – 73), and 43% (CI 33 – 71) for non-responders (P = 0.7). No clinical feature showed significant correlation with PB and BM molecular responses.This analysis shows that bcl-2 rearrangement in blood and bone marrow is frequently detected at staging, even in stage I disease. Absence of the bcl-2 rearrangement is related to a better EFS and the achievement of a molecular response in peripheral blood after therapy is associated with a better EFS.

Prognostic Value of BCL2 Determination in Follicular NHL Patients Treated with Alkylating Containing Regimen Alone or in Combination with Rituximab.

BCL2 over-expression is detected in follicular NHL patients in approximately 60%–80% at diagnosis. The prognostic value of the persistence of such molecular abnormality after treatment is still controversial: its presence could be associated with a shorter time to relapse and poorer survival, but there is no general agreement on that.Here we report clinical data retrospectively collected (1997 to 2001) on 60 newly diagnosed follicular NHL patients (39 female and 21 male) treated with two different schedules: 35 patients received alkylating containing regimen (CLB + PDN) as standard therapy, while 25 patients received Rituximab alone or in combination with CLB as first line treatment according to investigational studies.Median age was 55 years (range 28–76); 5 patients were considered in stage I, 14 in stage II, 15 in stage III and 26 patients in stage IV, according to Ann Arbor.The two groups of patients had similar clinical characteristics and prognostic factors.BCL2 determination was performed in 42/60 patients on bone marrow or peripheral blood, using PCR qualitative method. 25/42 patients (60%) were considered positive for BCL2 over-expression at diagnosis: 12/25 were in the group treated with chemotherapy and 13/25 in the Rituximab +/− chemotherapy arm.All patients were re-evaluated at the end of treatment both clinically and with molecular evaluation. Among patients who received CLB plus PDN, 24/35 (68%) achieved CR and 10/35 (28%) achieved PR. 7/12 (58%) BCL2 positive patients at diagnosis still presented positivity at the end of chemotherapy. With a median follow up of 36 months (range 6–134) 14 out 24 (58%) CR patients have relapsed. Among them only one was BCL2 positive at the end of therapy. Three additional relapsed patients were BCL2 negative after treatment.17/25 (68%) patients receiving Rituximab +/− chemotherapy achieved CR and 5/25 (20%) achieved PR. Only one patient out of 13 BCL2 positive at diagnosis presented BCL2 persistence at the end of therapy. With a median follow up of 36 months (6–71) only one BCL2 negative patient out of 17 CR has relapsed.In our experience no differences in terms of ORR was observed between the two groups of patients who received CT or Rituximab +/− chemotherapy.The higher rate of disappearance of BCL2 observed after Rituximab (98%) versus that recorded in patients receiving CT (48%) seems to confirm its more relevant clinical activity in disease eradication, and could be related to the lower incidence of relapse observed. However, the high degree of relapse observed in BCL2 negative patients after therapy could suggest no relationship between BCL2 persistence and the risk of subsequent relapse. The relatively short reported follow-up in the two groups of patients with such indolent disease, doesn't allow at present time any definitive conclusion about the prognostic value of BCL2 over-expression.

Bcl-2 protein expression is the strongest independent prognostic factor of survival in primary cutaneous large B-cell lymphomas

AbstractBcl-2 protein expression has been associated with poor prognosis in patients with noncutaneous diffuse large B-cell lymphomas. In primary cutaneous large B-cell lymphomas, the location on the leg, the round-cell morphology defined as the predominance of centroblasts and immunoblasts over large centrocytes, and multiple skin lesions were identified as adverse prognostic factors. The prognostic value of bcl-2 protein expression has not been studied in large series of patients. We evaluated 80 primary cutaneous large B-cell lymphomas collected by the French Study Group on Cutaneous Lymphomas. The prognostic value of age, sex, number of lesions, cutaneous extent, location, serum lactate dehydrogenase (LDH) level, B symptoms, morphology, and bcl-2 protein expression was studied. The overall 5-year specific survival rate was 65%. In univariate analysis, advanced age, multiple skin lesions (n = 48), location on the leg (n = 25), round-cell morphology (n = 32), and bcl-2 expression (n = 39) were significantly related to death from lymphoma. In multivariate analysis, bcl-2 expression (P = .0003), multiple skin lesions (P = .004), and age remained independent prognostic factors. The 5-year specific survival rates in bcl-2–positive and bcl-2–negative patients were 41% and 89%, respectively (P < .0001). A new prognostic classification of primary cutaneous B-cell lymphoma should be based primarily on bcl-2 protein expression rather than the location of skin lesions.

Proapoptotic genes BAX and CD40L are predictors of survival in transitional cell carcinoma of the bladder.

The purpose of the study was to investigate the effects of expression of a range of genes involved in apoptosis on outcome in bladder cancer. Immunohistochemistry was used to examine expression of BCL2, BAX, P53, CD40 and CD40L in archival tissues of patients included in various treatment trials for transitional cell carcinoma (TCC) of the bladder. Data were collected on 94 patients who first presented with either invasive or superficial bladder cancer. Median follow-up for alive patients was 83 months (m) (range 12–195 m). Median survival was 80 m (95% CI=56–128 m). Median survivals for the various markers were as follows: BAX-positive patients 110 m vs BAX-negative patients18 m (P=0.0002); CD40L-positive patients 95 m vs CD40L-negative patients 45 m (P=0.04); BCL2-positive patients 44 m and BCL2-negative patients 74 m, (P=0.64); CD40-positive patients 110 m and CD40 negative patients 45 m (P=0.12); and P53 positive patients 80 m and P53 negative patients 45 m (P=0.58). In conclusion, it was seen that overexpressions of BAX and CD40L are prognostic of better survival in TCC of the bladder. Our results also raise the possibility of the future development of CD40- and CD40 ligand-based immunotherapy for bladder cancer. This study links proapoptotic and antiapoptotic markers to overall survival.

Bcl-2 expression in head and neck cancer: an enigmatic prognostic marker

Purpose: The role of bcl-2 overexpression in cancer presents a paradox. In some tumor types, it is associated with favorable outcome, whereas in others the reverse is true. The purpose of this study was to explore the influence of bcl-2 in a large series of head and neck cancer patients treated in the CHART randomized trial. Methods and Materials: Histologic material was obtained from 400 patients; bcl-2 expression was assessed by immunohistochemistry as either positive or negative cytoplasmic staining. Results: Positivity of bcl-2 was recorded in 12.8% (9.5–16.5%, 95% confidence limits) of tumors. There were significant differences in positive tumors within different sites with nasopharynx showing the highest incidence (46.2%). A multivariate logistic regression analysis showed that bcl-2 was strongly associated with histologic dedifferentiation, as well as increasing N stage and female gender. In univariate analyses, bcl-2 positive patients had a lower locoregional relapse rate (RR 0.57, p = 0.02) and improved survival (RR 0.49, p = 0.004) compared to bcl-2 negative patients; this became more significant in multivariate analysis. Conclusion: These data demonstrate that bcl-2 overexpression is a marker of what is considered to be more advanced and aggressive disease yet it is associated with a more favorable outcome irrespective of the treatment schedule.