Proliferative Activity In Breast Cancer Research Articles

Could Apparent Diffusion coefficient (ADC) be used as an imaging marker for proliferative activity in breast carcinoma?

<p>Introduction. Multiparametric magnetic resonance mammography (mMRM) has an important role in detection, evaluation and follow-up of breast lesions. The aim of this study was to explore whether imaging parameters, in particular ADC, can be used as a biomarker of cell proliferation in breast cancer. Methods. This cohort-study included 67 lesions in 50 female patients who underwent mMRM on a 3T scanner. Percutaneous biopsies and surgical excisions were performed after imaging in period up to 3 weeks. The Ki67 index was assessed microscopically. Seven Ki67 categories were defined: 0–5%, 10–20%, 20–40%, 40–50%, 50–80% and over 80%. Methods of descriptive statistics were used; correlations were determined using Pearson’s correlation test. ROC curve was constructed and analyzed for determination of “cut-off” values for diagnostic potential. Statistical significance was set at p < 0.05. Results. Different subtypes of breast carcinomas were involved: ductal carcinoma (59.9%), lobular carcinoma (17.9%), metastatic carcinoma (10.5%), ductal carcinoma in situ (9%), and tubular carcinoma (3%). It was not possible to determine whether there was significant difference in values of ADC for histological subtypes of breast carcinoma because of small number of samples in some groups. The cut off value of ADC for breast carcinoma was 0.792 (sensitivity 98.6%, specificity 65.7%). There was no significant difference in values of ADC for categories of Ki67. There was no significant correlation between ADC mean and Ki67 for all histological subtypes of breast carcinomas (r = 0.156, p = 0.243). Conclusion. ADC cannot be used as a reliable imaging marker for proliferative activity in breast cancer.</p>

Ki 67: a Promising Prognostic Marker in Early Breast Cancer-a Review Article.

Ki67 index is considered to be a reliable indicator of the proliferative activity of breast cancer. Additionally, the Ki67 proliferative marker may play a role in assessing response to systemic therapeutic strategies and can act as a prognostic biomarker. But its limited reproducibility which stems from a lack of standardization of procedures, inter-observer variability, and preanalytical and analytical variabilities all have hampered the use of the Ki67 index in clinical practice. Currently, clinical trials have been evaluating Ki67 as a predictive marker for needing adjuvant chemotherapy in luminal early breast cancer patients receiving neoadjuvant endocrine therapy. But the inconsistencies existing in the estimation of the Ki67 index limit the utility of Ki67 in standard clinical practice. The purpose of this review is to evaluate the benefits and drawbacks of utilizing Ki-67 in early-stage breast cancer to prognosticate the disease and predict the risk of recurrence.

Prognostic Significance of Ki67 Expression By Immunohistochemistry (IHC) on Bone Marrow Biopsy in Newly Diagnosed Multiple Myeloma

Background: Multiple myeloma (MM) is known to be a heterogeneous disease encompassing several molecular subtypes that can be defined either by means of genetic or molecular testing. In the multigene tests, the ‘PR’ subtype enriched with proliferation-related genes has a substantial impact on the prediction of the risk of MM recurrence. Ki-67 is a nuclear protein involved in cell cycle regulation and is considered an indicator of proliferation. It is present in dividing cells during all active phases of the cell cycle (S, G1, G2 and mitosis phases) and is distinctly absent in resting or quiescent (G0) cells. Ki67 expression has been widely used as an index to evaluate the proliferative activity of breast cancer and lymphoma but the impact of Ki-67 expression on the clinical outcomes of patients with MM has largely been unexplored. This study aimed to investigate whether Ki-67 expression by IHC is an indicator of outcome in MM patients, especially those treated with novel therapies combined with or without autologous stem cell transplantation.Methods: The MM database was interrogated from January 2012-May 2017 for all newly diagnosed symptomatic MM subjects with Ki67 expression levels estimated by IHC on bone marrow biopsy specimens at diagnosis. Clinical features and outcomes were captured and compared between the cohorts of Ki67 high (Ki67-hi) subjects (Ki67 >5%) and Ki67 low (Ki67-lo) subjects (Ki67 ≤5%). Continuous variables were compared using median two sample tests, while incidences and proportions were compared using Fisher's exact tests. Survival distributions were estimated with Kaplan Meier techniques and compared between Ki67 cohorts using log-rank tests. Differences in overall survival (OS) and progression free survival (PFS) between the positive and negative cohorts were quantified with hazard ratios from Cox regression. Model selection was performed to determine adjusted hazard ratiosResults: A total of 125 MM subjects were identified with Ki67 values at diagnosis (26 Ki67-hi, 99 Ki67-lo). Gender, race, and age were similar between the two cohorts. There were no significant differences in beta-2 microglobulin (B2M) or LDH levels between the cohorts (p=0.236 and p>0.999, respectively); however, the median B2M in the Ki67-hi cohort was numerically higher than in the Ki67-lo cohort (4.6 vs 3.5 mg/L). A significantly greater proportion of Ki67-lo subjects had good risk cytogenetics (58.3%) compared to those who were Ki67-hi subjects (32.0%, p=0.035). There were no significant differences in the tested flow cytometry immunophenotype markers except for CD38, which was expressed in 87.9% of the Ki67-lo cohort compared to 53.9% of the Ki67-hi cohort (p<0.001). OS was significantly worse in the Ki67-hi cohort (HR: 3.12, 95% CI: 1.25, 7.75, p=0.010). Additionally, after adjusting for age and adverse risk, the risk of death in the Ki67-hi cohort is 4 times that of the Ki67-lo cohort (p=0.004, 95%CI: 1.58, 10.29). PFS was worse in the Ki67-hi cohort (HR: 1.86; 95% CI: 0.94, 3.65; p=0.069). Median follow-up time was 21.9 months (IQR 9.7 - 30.7 months).Conclusions: Our early results demonstrate that elevated Ki67 expression measured by IHC staining is associated with worse OS in newly diagnosed MM. IHC staining for Ki67 can potentially be utilized in economically constrained healthcare settings globally. Our current work focuses on standardization of Ki67 assessment and specification of its role as a dynamic biomarker of the treatment effectiveness in newly diagnosed MM. [Display omitted] DisclosuresVoorhees:Amgen: Speakers Bureau; Janssen: Consultancy, Speakers Bureau; Oncopeptides: Consultancy; Novartis: Consultancy; Takeda: Consultancy; Bristol-Myers Squibb: Consultancy; Celgene: Consultancy, Speakers Bureau. Usmani:Array BioPharma: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria, Research Funding; Skyline: Honoraria, Membership on an entity's Board of Directors or advisory committees; Onyx: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Takeda: Consultancy, Honoraria, Research Funding, Speakers Bureau; Amgen: Consultancy, Honoraria, Speakers Bureau; Novartis: Speakers Bureau; Millennium: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau; Sanofi: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Pharmacyclics: Honoraria, Research Funding. Bhutani:Amgen: Speakers Bureau; Prothena Therapeutics: Research Funding; BMS: Speakers Bureau; Takeda Oncology: Speakers Bureau.

Proliferative activity of various grades and types of breast carcinoma using AgNOR (Argyrophilic Nuclear Organizer Region) expression and its prognostic significance

A communal disease of the breast is breast cancer which happens mainly in women. Breast cancer is the most common cancer in women in both the developed and less developed world. The potential biological aggressiveness and behavior of breast neoplasm are of prognostic significance in tumor pathology and this can be assessed by measuring the tumor proliferative activity and malignant transformation. One of such markers is AgNOR (Argyrophilic Nuclear Organizer Region). A search of database sites such as PUBMED, Google Scholar, and Web of Science, as well as, other sources of literature available across public libraries was conducted to obtain information related to the topic. This review has presented a critical discussion on histologically assessing the proliferative activity of various grades and types of breast carcinoma using AgNOR expression and its prognostic significance. This exposé is anticipated to enlighten research scientists on specific management and assessing targets of tumors, which may merit prospective research into developing an appropriate mechanism for assessing the proliferative activity of breast cancer to help solve medical issues relating to cancers.

Enhanced E2F1 activity increases invasive and proliferative activity of breast cancer cells through non-coding RNA CDKN2B-AS1

Long non-coding RNAs have recently appeared as fundamental regulators of gene transcription in several biological processes, but only a few have known functional influences in the malignant transformation of breast cancer. CDKN2B-AS1 gene, also termed ANRIL, encoding a long non-coding RNA is located in the CDKN2B-CDKN2A gene cluster, loss of which is the most frequent alternation in several types of human malignancies. CDKN2B-AS1 is involved in the suppression of tumor suppressor genes (INK4a, ARF, and INK4b) and has been recognized as a direct target of E2F1. However, the roles of E2F1–CDKN2B-AS1 interaction in breast cancer have remained muchly mysterious. In this particular study, we reveal that both CDKN2B and CDKN2B-AS1 genes were differentially expressed in breast cancer cells in contrast to breast epithelial cells. Ectopic expression of E2F1 activated CDKN2B-AS1 but not CDKN2B expression. Lastly, overexpression of E2F1 improved the colony formation and migratory capacities of breast cancer cells. These results suggest that enhanced E2F1 activity increased invasive and proliferative activity of breast cancer but not breast epithelial cells possibly through up-regulating CDKN2B-AS1 transcript.

Abstract P2-03-07: Digital image analysis of Ki67 in hot spots is superior to alternative proliferation associated markers in breast cancer

Abstract INTRODUCTION Proliferative activity is one of the most important prognostic parameters in cancer. During the pathological examination of breast tumors, it is routinely evaluated by a count of the number of mitoses. Adding immunohistochemical stains of the nuclear protein Ki67 provides extra prognostic and predictive information. However, the currently used methods for both of these evaluations battle imperfections, primarily in reproducibility. In this study, we make an equally broad and detailed evaluation of mitoses, Ki67 and the more recently described Phosphohistone H3 (PHH3) in primary breast cancer using digital image analysis (DIA). Furthermore, we aim to investigate the prognostic and predictive value of proliferation-associated biomarkers in breast cancer stromal cells in relation to patient outcome. MATERIALS AND METHODS Two cohorts of primary breast cancer specimens (total n=297) with clinicopathological data including &amp;gt;10 years survival data, were sectioned and stained for Ki67, PHH3 and pancytokeratin (CKMNF116) and all glass slides were digitally scanned at x20. The DIA software used was the Visiopharm Integrator System (VIS) by Visiopharm A/S, Hoersholm, Denmark. VIS operates by a 'digital fusion' method that automatically excludes non-epithelial tissue restricting the analysis of the biomarkers (Ki67 and PHH3) to CKMNF116 positive cells. Both manual and DIA scores were compared for sensitivity and specificity for the gene expression based Luminal B versus A subtype, for high versus low transcriptomic grade as well as for their prognostic value in terms of Cox regression hazard ratios and breast cancer specific and overall survival. Further, we investigated whether the expression of Ki67 in the tumors' hot spots, invasive edges or as an average across all regions should be assessed for maximum power in relation to these outcomes. In addition, by inverting the DIA algorithm run by the VIS on the same cohorts, the expression of Ki67 and PHH3 was evaluated in the tumor stromal compartment. RESULTS Regardless of tumor region, DIA of Ki67 outperformed the other markers in sensitivity and specificity for gene expression subtypes and transcriptomic grade. In contrast to mitotic counts, tumors with high expression of Ki67 as defined by DIA, had significantly increased hazard ratio for all-cause mortality within 10 years from diagnosis. DIA of Ki67 was superior to manual Ki67 and PHH3 evaluations as well as to mitotic counts in terms of separation of patients with poor versus relatively good survival. Finally, we replaced the manual mitotic counts with DIA of Ki67 in hot spots as the marker for proliferation when determining histological grade. This increased the differences in estimated mean overall survival between the highest and lowest grades and added significantly more prognostic information to the classic Nottingham histological grade. CONCLUSIONS We conclude that digital image analysis of Ki67 in hot spots should be suggested as the marker of choice for proliferative activity in breast cancer. Citation Format: Robertson S, Stålhammar G, Wedlund L, Gholizadeh S, Lippert M, Rantaleinen M, Bergh J, Hartman J. Digital image analysis of Ki67 in hot spots is superior to alternative proliferation associated markers in breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P2-03-07.

Effectiveness of special training on achieving reproducible mitotic counts in malignant tumors

Background : M itotic count is the most commonly used method of assessing the proliferative activity of a tumor. It is used for classification, grading, prognostication of tumors and sometimes as a decision factor for treatment. Many variables related to procedure or person can influence the mitotic count. This study is designed to find out the effectiveness of special training in mitotic counting in achieving reproducible results. Methods : Sections from twenty cases of carcinoma breast formed the study material. Five junior residents were given the same twenty sections counted by the principal investigator for mitosis counting and original counts were recorded. Then they were instructed regarding mitotic figures using written material and graphic representations. All of them were asked to count the mitotic figures in the same section once again. The results of the two sets were compared. Paired t test and linear regression were the statistical tests used in analysis. Data storage and Analysis is done with EPIINFO software. Results: The mean standard mitotic counts obtained by principal investigator were 8.21. The mean original mitotic count obtained by residents was 15.79. After attending the training program the mean mitotic counts obtained by residents dramatically came close to that obtained by principal investigator ( 8.43 ) Conclusion : In trained hands, the mitotic counts are fairly reproducible and give results comparable to more sophisticated methods of determining proliferative activity in breast cancer. DOI: 10.21276/apalm.2017.999

Analysis of KI67 as a Prognostic Marker in Patients with Breast Cancer

BACKGROUND: Ki67 is used to evaluate the proliferative activity of breast cancer. Its role as prognostic marker is not well defined though. Studies have suggested that Ki67 could be used to select patients who would benefitiate more from adjuvant chemotherapy. In this study we tried to observe the correlation between Ki67 expression and the indication of chemotherapy during breast cancer treatment, and its correlation with other prognostic markers. MATERIALS AND METHODS: Chat review of 332 women with invasive breast cancer who were followed between 2000 and 2009 at a private service, in Salvador, Brazil. A 20% Ki 67 cut-off was selected. RESULTS: Considering high Ki 67 >20%, we were able to study two distinct groups of patients. We observed a positive and statistically significant outcome between high Ki67 expression and low expression of oestrogen and progesterone receptors, greater tumor size and histologic grade, higher percentage of lymphovascular invasion, lymph node involvement and distant metastasis, and indication of adjuvant chemotherapy. There were numeric but not statistically significant correlations between Ki67 expression and HER-2 positivity, recurrent disease and use of chemotherapy for local recurrence and metastasis. DISCUSSION: We observed an association between high Ki67 expression and markers that indicate worse prognosis in patients with breast cancer. Patients with Ki67 higher expression are more likely to use adjuvant chemotherapy. CONCLUSION: in this study, high Ki67 was related to prognostic markers as absence of hormonal receptors, greater tumor size and histologic grade, higher percentage of lymphovascular invasion, lymph node involvement and distant metastasis, and indication of adjuvant chemotherapy.

Metabolism of histamine in tissues of primary ductal breast cancer

Histamine performs an important role in the pathologic and physiologic aspects of the breast gland. Among monoamines, histamine demonstrates the greatest proliferative activity in breast cancer. The aim of the study was to evaluate histamine concentration in plasma and tissues of breast cancer dependent on the activity of histamine metabolism enzymes in neoplasmatic tissues of the breast gland. Ninety-five women aged 38 to 70 years were divided into 2 groups. The control group (group I) consisted of 30 healthy women. Group II consisted of 65 women with primary ductal breast cancer. The concentration of histamine in plasma was assessed by immunoenzymatic method. The concentration of histamine in cancerous tissues of the breast and the metabolism of histamine enzymes, specially histidine decarboxylase, decarboxylase of aromatic l-amino acids, N-histamine methyltransferase, monoamine oxydase B, and diamine oxydase, were determined using isotope technique. In the course of 24 hours, excretion of N-methylimidazoleacetic acid was evaluated by the methods of chromatography. The statistical analysis was made based on Statistica Pl Ed (StatSoft, Cracow, Poland, 1998). A significant increase in the concentration of histamine in plasma ( P < .01) and tissues of ductal breast cancers ( P < .001), and in the activity of histidine decarboxylase ( P < .01), aromatic l-amino acids ( P < .05), and histamine methyltransferase ( P < .05) was found. Activity of monoamine oxidase B ( P < .01) and diamine oxidase ( P < 0.01) and excretion of N-methylimidazoleacetic acid were significantly decreased compared with the control group ( P < 0.001). The conclusions are as follows: (1) Concentration of histamine in the plasma of women is dependent on the concentration of histamine in the tissues of ductal breast cancers. (2) The significant increase of histamine in cancerous tissues of ductal breast cancer could suggest the participation of this monoamine in the development of breast cancer. (3) The increase of histamine concentrations in ductal breast cancer tissues can be connected with the disturbances of the balance between synthesis and enzymatic inactivation of this monoamine. (4) The concentration of histamine in the plasma of women with ductal breast cancers is dependent on the number of involved lymph nodes and the grade of histologic malignancy.

Expression Profiles of Retinoblastoma Protein and p27Kip1 According to HER2 Amplification Status in Primary Breast Cancer

Purpose: Human epidermal growth factor receptor-2 (HER2)/neu amplification affects the cell proliferation through the modulation of multiple G1 cell cycle regulators in breast tumor cells. We performed this study to investigate whether retinoblastoma protein (pRB) and p27Kip1 were differently expressed according to the HER2 amplification status in human breast cancer. Methods: HER2 amplification was assayed by fluorescence in situ hybridization and the expression of cell cycle regulators were assayed by immunohistochemistry on 153 consecutive invasive breast cancers. The proliferative activity of breast cancer was analyzed according to the HER2 amplification and cell cycle protein expression status. Results: HER2 amplification was observed in 39 (25.5%) of 153 breast cancers. In the HER2 amplified breast cancers, the pRB expression was significantly increased (p=0.011) whereas there was no significant relationship between HER2 amplification and p27Kip1 expression. There was an inverse correlation between pRB expression and Ki67 labeling index in the HER2 amplified breast cancers (p= 0.036). In contrast, Ki-67 labeling index was significantly decreased as p27Kip1 expression increased in HER2 nonamplified breast cancers (p=0.028). In HER2 non-amplified breast cancers, we could not observe any association between the pRB expression and Ki67 labeling index. Conclusion: The proliferation of the breast cancers was associated with pRB expression in HER2 amplified tumors whereas it was associated with p27Kip1 expression in HER2 non-amplified tumors. The results of the current study indicate that the cell proliferative activity of the breast cancer is under different growth signal pathways according to HER2 amplification status.

Prognostic differences of World Health Organization–assessed mitotic activity index and mitotic impression by quick scanning in invasive ductal breast cancer patients younger than 55 years

The proliferation marker mitotic activity index is the strongest prognostic indicator in lymph node-negative breast cancer. The World Health Organization (WHO) 2003-defined procedure for determining WHO-mitotic activity index is often replaced by a quick scan mitotic impression. We evaluated the prognostic consequences of this practice in 433 T(1-3)N(0)M(0) lymph node-negative invasive ductal type breast cancers with long-term follow-up (median, 112 months; range, 12-187 months). Twenty-seven percent of the studied cases developed distant metastases, and 25% died of disease. Agreement between WHO-mitotic activity index (0-5 = 1, 6-10 = 2, >10 = 3) and mitotic impression (1, 2, 3) categories was 66% (kappa = 0.41), including 85% for category 1, 26% for category 2, and 52% for category 3. The WHO-mitotic activity index was a much stronger prognosticator than the mitotic impression, and the 10-year survival rates of the same categories (eg, mitotic activity index and mitotic impression category both 2) differed greatly. When grade was assessed by combining WHO-mitotic activity index or mitotic impression with the same values for tubular formation and nuclear atypia, grades disagreed in 18% of the cases. Deviation from the formal WHO-mitotic activity index assessment guidelines in breast cancer often results in erroneous prognosis estimations with therapeutic consequences and may explain why the prognostic value of proliferative activity in breast cancer is not always confirmed.

Clinical significance of proliferative activity evaluated by MIB-1 in the treatment and postoperative follow-up of early breast cancer

21054 Background: To evaluate a clinical significance of proliferative activity in breast cancer, we studied relationships between proliferative activity and recurrence rate, the time of recurrence or adjuvant therapy. Methods: We analyzed 2448 patients with primary breast cancer between 1987 and 2004 in the Kumamoto City Hospital, and 437 cases out of the patients developed recurrence. Furthermore, the rate of recurrence before and after 1999 when postoperative adjuvant therapy (such as CEF or Taxanes) was started as standard treatment was investigated. Proliferative activity was judged by immunostaining for MIB-1. The fraction of proliferating cells was classified into 3 degrees (=19%, 20–49%, 50%=). Median observation period was 70 months. Results: 1) Distribution of patients by proliferation was as follows; =19%:1215 cases(50%), 20–49%: 870 cases(35%), or 50%=: 363 cases(15%). There was a significant relationship between proliferative activity and tumor size, nodal status, ER, PgR, p53 or HER2 status. 2) Multivariate analysis for disease-free survival revealed that a proliferative activity was one of significant factors in node-negative and positive cases. Recurrence rate was 11.6% in cases with low proliferation and 31.0% in high proliferation. The mean period from operation to recurrence in cases with low proliferation was 50.2 months, whereas 19.9 months in high proliferation (p&lt;0.0001). Moreover, 74% of recurrent cases with high proliferation recurred within 2 years after operation, and there were few recurrences from the fifth year. 3) Patients with low proliferation frequently developed bone metastasis. In local recurrence, diffuse skin recurrence was often seen in cases with high proliferation. 4) The prognosis of patients in the later period (standard therapy group) was significantly better than that of patients in the earlier period, especially in high proliferation group. Conclusions: Proliferative activity might reflect aggressive behavior of breast cancer and predict the time of recurrence. The standard adjuvant therapy was effective in inhibiting early recurrence with high proliferation. It is important to take proliferative activity into consideration in the treatment and follow-up of breast cancer. No significant financial relationships to disclose.

Ki-67 as prognostic marker in early breast cancer: a meta-analysis of published studies involving 12 155 patients

The Ki-67 antigen is used to evaluate the proliferative activity of breast cancer (BC); however, Ki-67's role as a prognostic marker in BC is still undefined. In order to better define the prognostic value of Ki-67/MIB-1, we performed a meta-analysis of studies that evaluated the impact of Ki-67/MIB-1 on disease-free survival (DFS) and/or on overall survival (OS) in early BC. Sixty-eight studies were identified and 46 studies including 12 155 patients were evaluable for our meta-analysis; 38 studies were evaluable for the aggregation of results for DFS, and 35 studies for OS. Patients were considered to present positive tumours for the expression of Ki-67/MIB-1 according to the cut-off points defined by the authors. Ki-67/MIB-1 positivity is associated with higher probability of relapse in all patients (HR=1.93 (95% confidence interval (CI): 1.74–2.14); P<0.001), in node-negative patients (HR=2.31 (95% CI: 1.83–2.92); P<0.001) and in node-positive patients (HR=1.59 (95% CI: 1.35–1.87); P<0.001). Furthermore, Ki-67/MIB-1 positivity is associated with worse survival in all patients (HR=1.95 (95% CI: 1.70–2.24; P<0.001)), node-negative patients (HR=2.54 (95% CI: 1.65–3.91); P<0.001) and node-positive patients (HR=2.33 (95% CI: 1.83–2.95); P<0.001). Our meta-analysis suggests that Ki-67/MIB-1 positivity confers a higher risk of relapse and a worse survival in patients with early BC.

EXPRESSION OF THE G2M CHECKPOINT REGULATOR CYCLIN B1 IN BREAST CANCER AND ITS CORRELATION WITH PROGNOSTIC PARAMETERS

Background: Breast cancer is the most common malignancy in Egyptian females. A wealth of literature is available on various diagnostic and prognostic parameters including clinical, pathological and biological factors. The present study focuses on markers of cell cycle proteins as cyclin B1 and parameters of cell cycle kinetics, related growth factors and tissue polyamines in relation to hormone receptor status.Patients and methods: A total of 102 females made the subject of the study, 72 females with breast cancer, 14 with benign lesions and 16 healthy subjects serving as controls for serum arameters. Normal breast tissue served as control for breast cancer tissues. Tissue polyamines were measured by HPLC. yclin B1 was measured by Flow cytometry (FCM) and Western Blotting (WB). Insulin-like Growth factor 1(IGF1) was easured by enzyme immunoassay (EIA). Tumor markers (CEA and CA 15.3), estradiol and progesterone levels were measured by microparticle enzyme immune assay (MEIA).Results: Cyclin B1 was detected in 17/34 (50%) of the cases tested by FCM and only in 10% of the cases evaluated by WB. Cyclin B1 positivity tended to be associated with high grade tumors. The mean percentage of cyclin B1 positivity was higher in aneuploid tumors than in diploid ones, as well as in tumors with ≥ 10% S-phase fraction (SPF) compared to those with < 10 % SPF. Cyclin B1 positivity was significantly associated with patients of high G2/M phase (p=0.05). Cell cycle analysis showed that ER-ve tumors were significantly associated with high SPF. Tissues polyamines showed significantly increased levels when compared to both benign and normal groups (p< 0.001). Polyamines showed a significant association with ER status (p= 0.04), tumor size (p=0.04) and histopathologic type (p= 0.05). Serum IGF 1 showed insignificant increased level in breast cancer patients less than 45 yrs when compared to the age matched control group. Serum estrogen level showed no change in the breast cancer patients ≥ 45yrs compared to age matched control group, while progesterone level was significantly lower (p< 0.02) in breast cancer patients less than 45yrs old compared to age matched benign and control groups. Serum CEA and CA 15.3 showed significant increase in the breast cancer patients < 45yrs old compared to age matched benign and control groups (p=0.03 and 0.06, respectively).Conclusion: Polyamines proved value as markers of proliferative activity of breast cancer but their testing needs further standardization. Cyclin B1 is associated with aggressive tumors. Its assessment by FCM proved superior to western blotting. IGF1 might be better used to assess tumor recurrence and monitor cancer patients, and its use needs further evaluation. This study may contribute to understanding breast cancer cell growth biology in Egyptian females in view of improving therapeutic and chemopreventive strategies.

Prognostic implication of p53 protein expression in relation to nuclear pleomorphism and the MIB-1 counts in breast cancer

A close correlation of the p53 protein expression to nuclear pleomorphism and proliferative activity in breast cancer has been reported. The prognostic implications of p53 protein expression, however, in relation to nuclear pleomorphism and proliferative activity in breast cancer remain controversial. Nuclear pleomorphism and immunohistochemical reactivity for p53 protein and MIB-1 were evaluated on formalin-fixed paraffin-stored sections from 250 patients with breast cancer for whom the median follow-up duration was 6.4 years. p53 protein expression was positive in 66 (26.4%) of 250 cases. Nuclear pleomorphism was grade I or II in 169 (67.6%) cases and grade III in 81(32.4%)cases. The MIB-1 counts were more than 10% in 102 (40.8%) cases and less than 10% in 148 (59.2%) cases. There was a close correlation between p53 protein expression and nuclear pleomorphism (p<0.0001) and between p53 protein expression and MIB-1 counts (p<0.0001). Univariate analyses showed the 66 cases with positive p53 protein expression to have a significantly (p=0.0284) worse disease free survival (DFS) than the 184 cases with negative p53 protein expression. A multivariate analysis, however, on the variables including all of p53 protein expression, nuclear pleomorphism and MIB-1 counts indicated the MIB-1 counts (p=0.0041) as well as the lymph node status to be independently significant factors for DFS, while neither p53 protein expression nor nuclear pleomorphism were independently significant factors for DFS. The present study demonstrated that the p53 protein expression, nuclear pleomorphism and MIB-1 counts all demonstrated prognostic significance for breast cancer, while the most significant prognostic indicator among these three biological parameters was the MIB-1 counts.

The Relationship between Tumor Stage and the Expression of Multiple Drug Resistance, Neovascularization and Tumor Proliferative Activity in Breast Cancer

The Relationship between Tumor Stage and the Expression of Multiple Drug Resistance, Neovascularization and Tumor Proliferative Activity in Breast Cancer

Assessment of proliferative activity in breast cancer: MIB-1 immunohistochemistry versus mitotic figure count

The proliferative activity is one of the most important single prognostic parameters in breast cancer diagnosis and the time-honored measure of proliferative activity, the mitotic figure count, is an integral component of most combined prognostic scores. The detection of the cell cycle-specific antigens Ki-67, and the development of anti-Ki-67 antibodies, including the paraffin-reactive antibody MIB-1, have established immunohistochemical detection of cell cycle-specific antigens as a measure of proliferative activity in breast cancer diagnosis. The current study was performed to correlate mitotic figure counts with the proliferative activity as assessed by MIB-1 immunohistochemistry, taking into consideration the interobserver reliability of 5 pathologists in estimating mitotic figure counts. In 32 consecutive invasive ductal breast carcinomas, mitotic figure counts were performed independently by 5 pathologists. Mitotic activity was expressed as number of mitotic figures per 10 high-power fields and in a 3-tier score according to the Scarff Bloom Richardson system. Immunohistochemistry was performed using MIB-1 antibody, heat-induced epitope retrieval, and the standard avidin-biotin-immunoperoxidase method. MIB-1 immunohistochemistry was assessed in 3 representative 20 × fields by semiquantitative estimation (% of tumor cells positive) and by image analysis (number of MIB-1-positive cells/mm 2). We found a high degree of interobserver correlation among 4 experienced pathologists, in both mitotic figures per 10 high-power fields and the 3-tier scoring system. We observed significant, albeit weak, correlations between semiquantitative and quantitative MIB-1 immunohistochemistry and the number of mitotic figures per 10 high-power fields ( r between .36 and .53), but this significance was lost in 3 of the 5 observers when mitotic activity was expressed in the 3-tier scoring system. This study confirms mitotic figure counting in the hands of experienced pathologists as a valid, reproducible means of assessing proliferative activity in routine breast cancer diagnosis. The statistically significant, albeit only weak, correlation with MIB-1 immunohistochemistry is in agreement with results obtained by others and suggests that MIB-1 immunohistochemistry cannot be translated by a simple conversion factor into combined prognostic scores to replace the time-honored mitotic figure counts.

Value of immunohistochemical-determined oestrogen receptor status and proliferative activity in breast cancer: a retrospective study including patients treated by endocrine therapy at the time of recurrence

Abstract The oestrogen receptor (ER) status and the proliferative activity in breast carcinomas have proved to be of prognostic value. Because only half of the ER-positive patients respond to endocrine therapy it is hypothesized that it might be cases with positive ER status and high proliferative activity that fail endocrine treatment. Breast cancer patients with advanced/disseminated disease ( n = 109) were included in this study. No adjuvant systemic therapy was given, but when the patients developed recurrent disease they were all treated with endocrine therapy. The ER status and the proliferative activity (expressed as the ER index and MIB-1 index, respectively) were investigated retrospectively on sections from the primary tumour by immunostaining with monoclonal antibodies (anti ER-antibody and MIB-1) which are applicable on formalinfixed, paraffin-embedded tissue after microwave pretreatment. In ER-positive tumours there was no significant difference in response to endocrine therapy between tumours with high proliferative activity and tumours with low proliferative activity. This study shows that proliferative activity does not seem to predict response to endocrine therapy but indicates that proliferative activity is capable of revealing differences in the disease-free interval, irrespective of ER status.

Standardized Mitotic Counts in Breast Cancer Evaluation of the Method

Twenty-one pathologists and technicians participated in a study evaluating the variation present in mitotic counts for prognostication of breast cancer. The participants counted the mitotic figures in 20 breast cancer samples from ten high power fields (mitotic activity index, MAI, giving the results in mitotic figures per 10 fields) and also made a correction for field size and area fraction of the neoplastic epithelium to get the standardized mitotic index (volume fraction corrected mitotic index, or M/VV index, giving the result in mitotic figures per square mm of neoplastic epithelium). The difference in variation between the two methods was not big, but the standardized mitotic index (SMI) showed consistently smaller variation among all participants and different subgroups. Experienced pathologists had the highest variation in mitotic counts, and specially trained technicians, the lowest. The efficiency of the mitotic counts in grading (the grading efficiency) was used to evaluate the mitotic counts. In groups without special training for mitotic counts the mean grading efficiency was lower (experienced and training pathologists both on average had the potential to grade 88% of the cases correctly) than in the group specially trained for the purpose (trained technicians had the potential to grade 95% of the cases correctly). Among the specially trained technicians, the grading efficiency was of the same magnitude as the grading efficiency achieved in determining the S-Phase fraction of cells from paraffin embedded breast cancers by flow cytometry in different laboratories. The results suggest that special training is helpful in making mitotic counts more reproducible, and that in trained hands, the mitotic counts give results comparable to more sophisticated methods of determining proliferative activity in breast cancer.

Tissue polypeptide specific antigen (TPS): a new serum marker of proliferative activity in breast cancer

Tissue polypeptide specific antigen (TPS): a new serum marker of proliferative activity in breast cancer