Bcl-2 Expression In Patients Research Articles

Abstract P3-05-51: Patients with low expression of BCL 2 experience more late recurrence than patients who do not, which neutralized the protective effect of low BCL2 expression in patients with estrogen receptor positive breast cancer

Abstract Background) BCL2 is a key factor for the regulation of cellular apoptosis and its overexpression inhibits apoptotic cell death and activates cellular proliferation, resulting tumor progression. Although BCL2 are expected to be associated with an adverse prognosis in breast cancer, previous studies have reported inconsistent results and some reported even favorable features. The aim of this study is to investigate what characteristics of BCL2 yield this discrepant result in patients with breast cancer. Materials and methods) Female patients with breast cancer who completed primary treatment against breast cancer between 2003 and 2018 at Wonju Severance Hospital, Korea, were included. Clinocopathological characteristics including BCL2 expression were collected. Patients were categorized into two groups, BCL2 expression in more or less than 10% of tumor cells. Kaplan-Meier curves were generated to compare recurrence-free interval (RFI) and overall survival (OS). Result) The final cohort included 616 patients with a mean age of 54.79±11.2 (25-86) years. Patients with estrogen receptor positive breast cancer were more frequent in patients with tumor in which BCL2 was expressed in less than 10% of tumor cells (70.7% vs. 57.7%). Patients with tumor in which BCL2 was expressed in less than 10% of tumor cells showed better survival than their counterpart. Subgroup analysis according to the estrogen receptor and HER2 overexpression status was done. In patients with tumor in which HER2 was not overexpressed, patients with tumors in which BCL2 expressed less than 10% of tumor cells showed better survival(p=0.007). Survival difference maintained in patients with triple negative disease(p=0.010). However, in patients with estrogen receptor positive breast cancer, survival of two groups of patients with high and low BCL2 expression become not significant over time as late recurrence occurred (p=0.227). In contrast, patients with HER2 positive disease breast cancer showed worse prognosis than their counterpart when their tumor showed BCL2 expression in less than 10% of tumor cells. Conclusion) BCL2 overexpression showed different contribution to the patients’ survival according to the subtypes. Patients with HER2 overexpression showed better survival when BCL2 was overexpressed and patients with HER2 negative disease does not. First, similar survival was resulted from late recurrence in patients with estrogen receptor positive disease. Second, apoptotic capacity of tumor cells may not be major survival factor if tumor cells are exactly targeted. Citation Format: Hyang Suk Choi, Hany Hany Noh, In-Jeong Cho In-Jeong Cho, Seung Taek Lim, Jong-In Lee, Airi Han. Patients with low expression of BCL 2 experience more late recurrence than patients who do not, which neutralized the protective effect of low BCL2 expression in patients with estrogen receptor positive breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P3-05-51.

Cyclooxygenase-2 and Bcl-2 expression in patients with triple-negative breast cancer

Introduction. Triple-negative breast cancer (TNBC) is a rare type of breast cancer associated with lack of expression of estrogen and progesterone receptors and the HER2 protein. It is characterized by a poor outcome and chemotherapy resistance. Cyclooxygenase-2 (COX-2) is a constitutional enzyme responsible for prostaglandin synthesis, present in neoplastic cells and premalignant lesions. The B-cell lymphoma 2 (Bcl-2) protein is considered one of the most potent apoptosis-regulating agents, assuring body homeostasis. Material and methods. The aim of the present study was to evaluate the immunohistochemical (IHC) profile of COX-2 and Bcl-2 expression in patients suffering from TNBC in order to obtain more detailed data on additional factors nega­tively influencing TNBC outcome. The IHC evaluation of COX-2 and Bcl-2 expression among 21 women with diagnosis of TNBC was performed. Results. The most common histological subtype was invasive ductal cancer of no special type. COX-2 was present in all examined samples with moderate to strong expression detected in 20 of 21 cases. There was a positive corre­lation between histological grade (G) and COX-2 expression (p = 0.002). Bcl-2 was present in all examined samples. The analysis showed that tumours presenting highly positive expression of Bcl-2 accounted for the majority of examined cases (57.2%). Conclusions. The achieved results might lead to a conclusion that COX-2 and Bcl-2 high expression in TNBC may be linked to tumour aggressiveness and poor overall survival. However, before their consideration as additional markers to be used in routine histological examinations and breast cancer grading, it will be necessary to undertake further studies.

Decreased expression of TRIM3 gene predicts a poor prognosis in gastric cancer.

Tripartite motif-containing 3 (TRIM3) is a member of the TRIM protein family which is known to be involved in development of numerous tumor types. However, the prognostic role of TRIM3 in gastric cancer (GC) remained to be clarified. The aim of this study was to evaluate the expression pattern and prognostic significance of TRIM3 gene and its relationship with β-catenin, CyclinD, and BCL2 expression in patients with GC. A total of 40 fresh primary gastric cancer tumors and their matched adjacent noncancerous tissues were selected from the First Affiliated Hospital of Mashhad University. Real-time quantitative RT-PCR was performed to evaluate differences in TRIM3 expression in GC and normal tissues. The correlation between TRIM3 expression level and patients' overall survival, some clinicopathological variables, and β-catenin, CyclinD, and BCL-2 genes expression level were also studied. Moreover, patients were divided in two groups according to the TRIM3 expression levels: low and high. Compared to noncancerous tissues, TRIM3 expression in GC tissues was significantly increased (fold change = 1.58). Kaplan-Meier survival analysis revealed a significant difference of patient survival according to TRIM3 expression status (P = 0.012). Low TRIM3 expression was associated with shorter overall survival and was an independent predictor for poor prognosis in GC patients (HR, 1.25; 95%CI, 1.02-1.60; P = 0.045). Expression of TRIM3 was negatively correlated with expression of β-catenin, BCL-2, and CyclinD as genes for proliferation, apoptosis, and the cell cycle in GC patients. This study demonstrates that decreased level of TRIM3 mRNA expression is associated with poor prognosis in patients with gastric cancer. TRIM3 may play a protective role in gastric cancer by relieving the effects of cancer progressive genes and could be considered for further investigations as a prognostic biomarker.

1806P - Salivary cytokines and oral mucosa cells apoptosis in patients during hematopoietic cell transplantation: Possible relationship with oral mucositis

BackgroundApoptosis has a central role in the process of oral mucositis and can be sustained by proinflammatory cytokines. Some studies have demonstrated increased levels of tumor necrosis factor alpha (TNF-α), interleukin 1β (IL-1β) and interleukin 6 (IL-6) in saliva after radiotherapy and chemotherapy, which has been associated to severe oral mucositis. However, little is known about the role of these cytokines in the pro and anti-apoptosis process of oral cells exposed to antineoplastic agents. The aim of this study was to verify whether there was an association of salivary cytokines with oral cells exhibiting apoptosis or anti-apoptotic Bcl-2 expression in patients who experienced oral mucositis during hematopoietic cell transplantation (HCT). MethodsWe collected saliva and oral mucosa cells from 77 HCT patients before the conditioning (T0), during the neutropenia period (T1), and after the marrow recovery (T2). We quantified the oral exfoliated cells exhibiting TUNEL positivity and immunocytochemical expression of Bcl-2. We also determined the salivary levels of TNF-α, IL-1β, and IL-6 in the three periods. ResultsThe number of cells exhibiting positivity to TUNEL (p=0.021) and Bcl-2 (p=0.020) increased from T0 to T2. The levels of TNF-α increased significantly from T0 to T1 (p<0.001). There were no significant alterations of IL-1β and IL-6 comparing the three periods. There was a significant positive correlation between TNF-α and TUNEL in T0 (rho=0.277, p=0.033) and T1 (rho=0.451, p=0.020). Bcl-2 expression in T1 (rho=--0.335, p=0.048) was negatively correlated with time duration of oral mucositis. ConclusionsOral cells exhibited apoptosis even after the marrow recovery. Oral cells apoptosis was associated with salivary TNF-α during the neutropenia period. Oral cells also exhibited high expression of antiapoptotic protein Bcl-2, which was inversely proportional to the duration of oral mucositis, suggesting the activation of a surveillance mechanism in the oral cells. Alterations on salivary TNF-α during the HCT may have a putative role in the apoptosis presented in oral mucositis. Legal entity responsible for the studyHospital Israelita Albert Einstein. FundingSão Paulo Research Foundation (2016/03650-4) and AmigoOH - Brazil. DisclosureAll authors have declared no conflicts of interest.

BCL-2 Expression in Patients with Non Hodgkin's Lymphoma with Hepatitis C

The mechanism of lymphoma genesis in hepatitis C virusrelated, B cell NHL is unknown. It has been suggested that HCV may induce B cell clonal proliferation and BCL-2 gene overexpressionin chronicHCV patients. Aim of the Work: Is to evaluate the expression of BCL2 gene in patients with NHL with HCV infection. Methods: The study included 92 patients, divided into 52 patients with B-cell NHL with chronic HCV infection and 40 patients with B-cell NHL without HCV infection who attended the National Cancer Institute, Cairo University, during the period from March 2012 to May 2014. Lymph node biopsies from all cases were studied for Bcl-2 expression using tissue microarray and immunohistochemistry techniques. Results: BCL-2 was expressed in 32.7% of cases with Bcell NHL with hepatitis C and in 20% of cases of HCV negative NHL by immunohistochemical method and the Bcl-2 gene was up-regulated more than 2 folds in NHL HCV+ve patients than in NHL HCV–ve group by tissue microarray technique with statistically significant difference. Conclusion: Bcl-2 is over expressed in NHL with HCV infection more than in NHL without HCV and could play a role in lymphoma genesis and future therapeutic strategies.

Prognostic value of Bcl-2 expression in patients with non-small-cell lung cancer: a meta-analysis and systemic review

ObjectiveB-cell-lymphoma-2 (Bcl-2) is a proto-oncogene that plays an important role in the regulation of apoptosis and cell survival. However, there are much conflicting data in the literature concerning the association between Bcl-2 and prognosis in non-small-cell lung cancer (NSCLC). There is little in the way of meta-analysis focused on Bcl-2 and its effect on NSCLC prognosis. This study was performed to provide an assessment of whether expression levels of Bcl-2 are associated with prognosis in patients with NSCLC.Materials and methodsWe searched PubMed, the Cochrane Library, and China National Knowledge Infrastructure for all eligible studies. The combined hazard ratios (HRs) and their corresponding 95% confidence intervals (CIs) in terms of overall survival were evaluated.ResultsFifty published studies including 6,863 patients with lung cancer were included in this meta-analysis. Overall, Bcl-2 was expressed in 33% of the NSCLC tumors studied. Our analysis indicates that NSCLC patients with Bcl-2-positive expression have a better prognosis than those with Bcl-2-negative expression in both Asian and non-Asian study populations (HR 0.79, 95% CI 0.72–0.87, P<0.00001). However, Bcl-2-positive expression seems to have no significant impact on survival of stage I NSCLC patients.ConclusionOur results indicated that Bcl-2 might be a useful prognostic marker for NSCLC generally. Larger clinical trials are needed to confirm the prognostic value of Bcl-2 in stage I NSCLC.

High expression of Bcl-2 protein predicts favorable outcome in non-small cell lung cancer: evidence from a systematic review and meta-analysis.

The prognostic value of Bcl-2 protein expression in non-small cell lung cancer (NSCLC) is under debate. We therefore systematically reviewed the evidence for Bcl-2 protein effects on NSCLC survival to elucidate this issue. An electronic search in Pubmed and Embase complemented by manual searches in article references were conducted to identify eligible studies to evaluate the association between Bcl-2 protein expression and overall survival (OS) as well as disease free survival (DFS) of NSCLC patients. Combined hazard ratios (HRs) with corresponding 95% confidence intervals (95%CIs) were pooled using the random-effects model. A total of 50 trials (including 52 cohorts) encompassing 7,765 patients were pooled in the meta-analysis regarding Bcl-2 expression and OS of NSCLC patients. High expression of Bcl-2 protein had a favorable impact (HR=0.76, 95%CI=0.67-0.86). In the group of Bcl-2 expression and DFS, 11 studies including 2,634 patients were included. The synthesized result indicated high expression of Bcl-2 protein might predict good DFS (HR=0.85, 95%CI=0.75-0.95). Our present meta-analysis demonstrated favorable prognostic values of Bcl-2 expression in patients with NSCLC. Further prospective trails are welcomed to validate the utility of assessing Bcl-2 in NSCLC patient management.

Positive immunohistochemical expression of bcl-2 in hormone-independent breast carcinomas is associated with a greater lymph node involvement and poor outcome

To study the immunohistochemical expression of bcl-2 in patients with hormone-independent breast infiltrating ductal carcinomas (IDC) and its possible association with other clinico-biological parameters and outcome. Our study group included 72 females with hormone-independent (ER and PgR negative) infiltrating ductal breast carcinomas. Age, tumor size, axillary lymph node involvement (N), distant metastasis and histological grade, as well as the immunohistochemical expression of Ki67, p53 and androgen receptor (AR), were analyzed. We follow up 57 patients during a period of time ranged between 20 and 193 months (80.2 ± 58.3; median 78 months). Of all IDCs included in our study, 18 were ER-/PgR-/bcl-2+ and 54 ER-/PgR-/bcl-2-. The percentages of slightly bcl-2-positive (+) and bcl-2-strong positive (++) cases were 25 and 19 %, respectively, values lower than those observed in ER+/PgR+ tumors (79.3 and 86.8 %, respectively). Breast IDC with positivity (+) for bcl-2 showed, exclusively, greater lymph node involvement higher than 3 nodes (N+ >3) (p 0.021) and a great number of deaths due to the tumor (p 0.011). Same results were obtained when we compared bcl-2-negative and bcl-2-strong positive (++) subgroups. Our results led us to consider that the positive (+ or ++) immunohistochemical expression of bcl-2 in hormone-independent (ER and PgR negative) breast carcinomas is associated with greater axillary lymph node involvement and a greater number of deaths in the follow-up, being these data opposite to that observed in hormone-dependent tumors.

Prognostic Value of p53 and bcl-2 Expression in Patients Treated with Breast Conservative Therapy

Prognostic value of p53 and bcl-2 expression on treatment outcome in breast cancer patients has been extensively evaluated, but the results were inconclusive. We evaluated the prognostic significance of these molecular markers in patients treated with breast conserving surgery and radiotherapy. One hundred patients whose immunostaining of p53 and bcl-2 expression was available among 125 patients who underwent radiotherapy after breast conserving surgery and axillary lymph node dissection were enrolled into this study. Eighty-seven patients also received adjuvant chemotherapy and/or hormonal therapy. Conventional clinicopathologic variables and treatment-related factors were also considered. The 5-yr loco-regional relapse-free and distant metastasis-free survival rates were 91.7% and 90.9%, respectively. On univariate analysis, age, T stage and the absence of bcl-2 & estrogen receptor (ER) expression were associated with loco-regional relapse-free survival. When incorporating these variables into Cox proportional hazard model, only bcl-2(-)/ER(-) phenotype was an adverse prognostic factor (P=0.018). As for the distant metastasis-free survival, age, T stage, and p53 expression were significant on univariate analysis. However, p53 expression was the only prognosticator on multivariate analysis (P=0.009). A bcl-2(-)/ER(-) phenotype and p53 expression are useful molecular markers predicting loco-regional relapse-free and distant metastasis-free survival, respectively, in patients treated with breast conserving surgery and radiotherapy.

The Impact of Selected Apoptotic Markers and Adhesion Molecules on the Treatment Outcome of Chronic Lymphocytic Leukemia

Brief Abstract We assessed the impact of selected apoptotic markers and adhesion molecules on the treatment outcome. Thirty newly diagnosed patients and 10 healthy subjects were studied. Proapoptotic markers (BCL2, p53, and CD95) and adhesion molecules (CD18, CD54) were assessed by flow cytometry before and after treatment. p21 mRNA was assessed by RT-PCR. Induction was done with the CVP regimen. Full Abstract The study included 23 men and 7 women. Median age was 66 years (range, 53-79 years). There were increases of p53 and BCL2 and reductions of CD18 and CD54 in cases compared to control. CD95 did not show significant increase compared to control. There were reductions in BCL2 after (versus before) treatment and increases in CD18 and CD54 after (versus before) treatment. Evaluable patients showed 11 complete remission (CRs; 39.3%), 7 (25%) partial remissions, 6 progressive diseases (21.4%), and 4 stable diseases (14.3%). The observation after treatment was 2 to 13 months. Median time to disease progression (TTP) of the responding patients was 9 months. Death was reported in 1 case. There was an increase in p53 and BCL2 expression in patients who did not achieve CR compared to those with CR and an increase in CD18 level in CR more than no-CR cases. No significant differences were found regarding the expression of CD95 and CD54 between both groups. Comparing the level of expression of the studied markers at initial presentation, there were increases in p53 and decreases in CD18 level in patients with no CR compared to those with CR. There was positive correlation between the expression of p53 and BCL2 and negative correlation between the p53 and CD18 and between BCL2 and CD18. There was significant negative correlation between TTP and BCL2 levels after treatment. Selected cases for p21 mRNA assessment showed negative correlation between p21 mRNA and p53 levels before treatment and decrease in p21 level was associated with poor prognosis and bad response to treatment. In conclusion, measurement of p53 and CD18 level can define a group of patients with aggressive disease who may require more aggressive therapy. BCL2 level after treatment is correlated with TTP. Acknowledgment The study was conducted at NCI-Cairo between 2004 and 2007.

Prognostic Value of p53 and bcl-2 Expression in Patients Treated With Breast Conservative Therapy

Prognostic Value of p53 and bcl-2 Expression in Patients Treated With Breast Conservative Therapy

Expression of apoptosis regulatory proteins p53 and Bcl‐2 in skin of patients with chronic renal failure on maintenance haemodialysis

Chronic renal failure results in multi-organ system derangement including cardiovascular, gastrointestinal, neurological, endocrinal, blood and dermatological abnormalities. Maintenance of skin homeostasis requires a delicate balance between proliferation, differentiation and apoptosis. p53 and Bcl-2 proteins play a central role in the regulation of apoptosis. Evaluation of the expression of apoptosis regulatory proteins p53 and Bcl-2 in apparently normal skin of patients, with chronic renal failure on maintenance haemodialysis, with respect to their role in the apoptotic process. Biopsy specimens were obtained from 10 patients with chronic renal failure on maintenance haemodialysis, as well as seven age-matched control subjects. Computer-assisted image analysis was employed to measure epidermal thickness in H&E-stained sections. Immunoperoxidase technique was also used to demonstrate p53 and Bcl-2 proteins and the TUNEL technique for detection of apoptotic cells in these specimens. The mean epidermal thickness was significantly higher (P < 0.0001) in patients than controls. Meanwhile, no apoptotic cells were detected in the epidermis of patients. On the other hand, a statistically significant difference was observed in both p53 (P = 0.0001) and Bcl-2 expression (P = 0.0003) when comparing patients and controls. Expression of p53 (2.74 +/- 0.84) and Bcl-2 (3.45 +/- 1.35) proteins was higher in skin samples obtained from patients with chronic renal failure and on maintenance haemodialysis than those from control cases (0.5 +/- 0.96 and 0.8 +/- 0.6, respectively). Moreover, Bcl-2 expression in patients was observed in basal as well as squamous cell layers of skin, whereas in control subjects it was confined to the basal cell layer only. These findings suggest that an alteration in the proliferation/apoptosis balance may be present in the skin of such patients.

The Impact of High-dose Sodium Selenite Therapy on Bcl-2 Expression in Adult Non-Hodgkin’s Lymphoma Patients: Correlation with Response and Survival

The present study was undertaken to explore the effect of administration of high doses of sodium selenite on the expression of Bcl-2 in patients with non-Hodgkin's lymphoma (NHL). Fifty patients with newly diagnosed NHL were randomly divided into two groups. Group A-I received standard chemotherapy whereas group A-II received adjuvant sodium selenite 0.2 mg kg-1 day-1 for 30 days in addition to chemotherapy. Enzyme-linked immunosorbent assay was used to assess Bcl-2 at the time of diagnosis and after therapy in the two groups. Sodium selenite administration resulted in significant decline of Bcl-2 level after therapy in group A-II (8.6 +/- 6.9 ng/ml vs 3 6.9 +/- 7.9 ng/ml, P < 0.05). Also, complete response reached 60% in group A-II compared to 40% in group A-I. Significant increase in CD4/CD8 ratio was noticed in group A-II compared to group A-I after therapy (1.45 +/- 0.36 vs 1.10 +/- 0.28 p 0.04). Overall survival time in months was significantly longer in complete remission patients in group A-II (21.87 +/- 1.41) compared to group A-I (19.70 +/- 1.95) (p = 0.01). It is concluded that sodium selenite administration at the dosage and duration chosen acts as a down regulator of Bcl-2 and improves clinical outcome.

In Vitro Colony Growth, RAS Genes Sequencing, and Expression of Bcl-2 and Bax Proteins in Chronic Myelomonocytic Leukemia.

Chronic myelomonocytic leukemia (CMML) is a heterogeneous hematological malignancy, which shows features of both myelodysplastic and myeloproliferative disorders. It has poor prognosis due to the lack of effective treatment and very little is known about its pathogenesis. A point mutation in the RAS genes is detected in 20 to 35% of patients with CMML, with higher percentage among patients with proliferative variant of the disease (MP-CMML, WBC > 13 x 109/L). The expression of Bcl-2 family proteins, which are key regulators of the mitochondrial-mediated pathway of apoptosis, have been reported as being altered in MDS and other hematological disorders. However, the role of these proteins has not been systematically investigated in CMML.In this study, we evaluated by Western Blotting the expression of anti-apoptotic Bcl-2 and pro-apoptotic Bax proteins in peripheral blood mononuclear cells (PBMNC) from 21 CMML patients and 8 age-matched healthy controls. We also investigated the presence of point mutations within codons 12, 13, and 61 of N- and K-RAS by sequencing analysis, and assessed in vitro colony growth of PBMNC with and without growth factors (GFs) [SCF, GM-CSF, IL-3, Epo]. Our series included 10 males and 11 females, with a median age of 74 years. Median WBC value was 9.2 x 109/L (range 3.2 to 17.2), while differential showed median monocyte values of 27.5% and 2.0 x 109/L. Six out of the 21 patients had MP-CMML. All patients but one had normal karyotype. A point mutation within N-RAS (G12D) was detected in two patients, both with MP-CMML. Spontaneous growth was observed both in normal controls and in CMML (median 19 and 14 CFU-GM/105 MNCs, respectively), with a higher number of CFU-GM in the MP-CMML than in the dysplastic subgroup (MD-CMML) (46 vs 12). After addition of GFs, we observed an increase of colony number in all groups. Interestingly, in vitro colony growth was remarkably higher in the two patients with mutant N-RAS than in patients with wild type RAS, especially after the addition of GFs (200 vs 57, p <0.05). The expression of Bax was higher in CMML than in the control group (3.81 vs 1.39, p = 0.07), whereas Bcl-2 was lower (0.88 vs 1.41, p = .16), even though these differences were not statistically significant. No differences were observed between subgroups of patients with MD- and MP-CMML. When the Bcl-2/Bax ratio was calculated, we observed lower values in CMML than in normal controls (0.27 vs 1.08, p <0.01). In the two patients with mutated RAS, Bcl-2 expression was significantly higher than in patients with wt-RAS (2.07 vs 0.75, p <0.05). Noteworthy, Bcl-2 expression in patients with mutant RAS was also higher than in the control group. In conclusion, our data suggest a deregulation of mechanisms controlling apoptosis in CMML, with no significant differences between MD and MP variants. The finding of higher Bcl-2 expression in patients with mutant RAS suggests a possible cooperation of this anti-apoptotic protein with RAS-activated intracellular pathways, warranting further confirmation in larger CMML series. In agreement with previous reports, we observed higher in vitro spontaneous growth in MP-CMML than in MD-CMML. Hypersensitivity to GFs (possibly GM-CSF) in cells from patients with mutant RAS is suggested and merits further investigation.

Prognostic Significance of BCL-2 Expression in Patients with Multiple Myeloma.

Background: multiple myeloma (MM) remains an incurable disease that accounts for approximately 10% of all hematologic cancers. Patients (pts) with MM treated with conventional chemotherapy have a median overall survival of 36–42 months. Various adverse prognostic factors have been described in MM, including elevated B2 microglobulin, hypoalbuminemia, cytogenetic abnormalities of chromosome 13 (13q14 del, monosomy 13), and elevated CRP among others. BCL-2 mediates anti-apoptotic pathways and its expression has been implicated with lack of response to certain chemotherapeutic agents and unfavorable prognosis in different human malignancies.Objective: to evaluate the prognostic significance of BCL-2 expression in patients with MM.Methods: adult pts who were diagnosed with MM at our institution since 1/1/90 were retrospectively analyzed. Expression of BCL-2 was determined by immunohistochemistry (IHC) staining of deparafinized bone marrow specimens. Median survival was calculated for BCL-2 (+) and BCL-2 (−) groups. Survival curves were estimated according to the Kaplan-Meier method, and were compared with the use of the log-rank test. Median survivals were also compared by using Wilcoxon-Mann-Whitney test. Two-tailed alpha level of 0.05 was considered statistically significant. SAS® software (Cary, NC) was used for statistical analysis.Results: sixty-six patients were initially identified. Forty-four pts were excluded because bone marrow samples were not available for IHC staining. Twenty-two pts (11 males, 11 females), with a median age at the time of diagnosis of 64 years (range: 46 to 83 years) were included for this analysis. Fourteen pts (64%) had positive BCL-2 bone marrow specimens, including 4 mild, 3 moderate, and 7 strong staining, while for 8 pts (36%), bone marrow specimens did not stain for BCL-2. Twelve pts (86%) from BCL-2 (+) group and 4 pts (50%) from BCL-2 (−) group died during the period of follow-up; 8 pts are still alive at the time of this analysis, 23 to 71 months (median: 39 months) after diagnosis. The median survival among BCL-2 (+) pts was 35 months (95% CI: 24 – 69 months) and among BCL-2 (−) pts was 47 months (95% CI: 13 months - N/A). Overall survival did not differ between the two groups (p-values for log-rank and Wilcoxon-Mann-Whitney tests, 0.8256 and 0.7072, respectively).Conclusion: although our study did not show correlation between BCL-2 expression and overall survival, median survival among BCL-2 (−) pts with MM was 34% longer than BCL-2 (+) pts. The authors recognize the limitations associated with this analysis including the retrospective nature of the study, small sample size, and exclusion of a large number of patients. This study highlights the need for prospective evaluation of the prognostic significance of BCL-2 expression among pts with MM.

133 A study examining epidermal growth factor receptor, p53, and Bcl-2 expression in patients with stage III and IV squamous cell carcinoma of the head and neck treated with concomitant chemo-radiotherapy

133 A study examining epidermal growth factor receptor, p53, and Bcl-2 expression in patients with stage III and IV squamous cell carcinoma of the head and neck treated with concomitant chemo-radiotherapy

Bcl-2 expression and other clinicopathologic parameters in uterine leiomyosarcoma

The bcl-2 protein, which is involved in cell-cycle regulation and apoptosis, seems to play an important role in the growth of tumors. Until recently, only one study described the frequent expression of bcl-2 in uterine leiomyosarcomas (LMS) and indicated its important prognostic role. Thus, the aims of the present study were to examine bcl-2 expression in patients with uterine LMS and to determine if the expression of this protein would influence the prognosis. In addition, a possible association between bcl-2 expression and various clinicopathologic parameters was evaluated. Bcl-2 expression was investigated using immunohistochemical techniques on paraffin-embedded tissue from 21 patients with uterine LMS. The immunohistochemical findings were correlated with different clinicopathologic parameters (FIGO stage, age, vascular-space involvement and recurrence of disease) of the patients. Bcl-2 was expressed in 57% (12/21) of uterine LMS: 3/12 (25%) stained weakly positive, 3/12 (25%) stained moderately positive and 6/12 (50%) stained strongly positive. Nine (43%) cases were negative. Evaluation of the correlation between bcl-2 expression and several clinicopathologic parameters identified a significant negative correlation between vascular-space involvement and bcl-2 expression (p=0.04). The relationship between bcl-2 expression and FIGO stage, age and recurrence of disease did not reach statistical significance. A univariate Cox model revealed significantly longer overall survival in women with bcl-2-positive LMS compared with women with bcl-2-negative LMS (p=0.03). Significant negative correlation was found between bcl-2 expression and vascular-space involvement and improved survival rates in patients with bcl-2-positive tumors. Though limited by the small number of patients, these data indicate the role of bcl-2 expression in uterine LMS.

Apoptotic rate in patients with myelodisplastic syndrome treated with modulatory compounds of pro-apoptotic cytokines.

Excessive apoptosis has a central role in ineffective hematopoiesis in myelodysplastic syndrome (MDS). The aim of the study was to quantify apoptosis and Bcl-2 expression in patients with MDS and to use these parameters in the evaluation of treatment efficacy with compounds modulating proapoptotic cytokines. Bone marrow (BM) samples from eight MDS patients were studied: four with refractory anemia and four with refractory anemia with ringed sideroblasts. Two patients with Hodgkin disease without BM determination were studied for control. Therapy consisted in administration of pentoxyphylline, dexamethasone and ciprofloxacin. Biochemical assay of apoptosis and Bcl-2 was performed using annexin V-biotin conjugate antibody and anti-human Bcl-2 antibody respectively, followed by streptavidine-peroxidase conjugate, and peroxidase substrate. Ultrastructural investigation of BM samples was performed with standard electron microscopy techniques. Most of BM hematopoietic cells in the MDS patients had ultrastructural features of various stages of apoptosis including chromatin condensation and margination, cytoplasm condensation and budding of nuclear and plasma membranes to produce apoptotic bodies. Bcl-2 expression showed an inverse correlation with the rate of the apoptotic process. Periodic evaluation of these two parameters has shown an increase of Bcl-2 expression and a decrease of apoptotic rate in patients who had responded to the treatment. Response to the treatment was appreciated in accordance with their transfusion needs. Treatment efficiency diminished in time. The rate of apoptosis was inversely correlated with the level of Bcl-2 expression. These results confirm the importance of the apoptotic process evaluation in monitoring MDS treatment.

Haematopoietic response and bcl-2 expression in patients with acute myeloid leukaemia.

Bcl-2 expression, the number of apoptotic cells and the growth and differentiation of early bone marrow progenitor cells were studied in patients with confirmed diagnosis of acute myeloid leukaemia (AML). Bone marrow cells from normal individuals were used as controls. We observed an increased percentage of bcl-2-mononuclear bone marrow cells expression in AML patients in relation to controls (p =0.002). Accordingly, the number of apoptotic cells was reduced (p = 0.001) and there was a negative correlation between bcl-2 expression and the number of apoptotic cells (r=-0.664, p<0.001). In addition, bcl-2 expression was significantly increased in the chemotherapy resistant group in relation to the responsive group (p = 0.03). Lower rate of survival was observed in the group of AML patients with autonomous proliferation (p=0.01). These results suggest that a high bcl-2 expression and the presence of autonomous proliferation are related to a poor prognosis in AML.

Prognostic significance of tumor apoptosis, p53 and bcl-2 expression in patients with advanced epithelial ovarian cancers

Prognostic significance of tumor apoptosis, p53 and bcl-2 expression in patients with advanced epithelial ovarian cancers