Prognostic value of Ki 67/MIB1 automated and quantitative immunolabelling in primary operable breast carcinomas

The prognostic significance of Ki 67/MIB1 immunohistochemical assays (ICAs) was investigated in optimal technical conditions in 139 breast carcinomas. Automated ICAs (immunoperoxidase/Ventana device) was performed on frozen sections. Immunoprecipitates were quantified by computerized analysis (SAMBA) of digitized microscopic images. Mean positive surfaces (%) and quantitative immunocytochemical (QIC) index were correlated to the patients survival (8-year survival). The results showed that Ki 67/MIB1 large surfaces (cutpoint, 20%) and high QIC index (cutpoint, 12) correlated with poor overall survival (Kaplan Meier, log rank test, p=0.011 and p=0.037, BMDP software). In node positive, but not in node negative patients, large Ki 67/MIB1 surface (cutpoint, 20%) and high QIC index (cut-off 12) correlated with the overall patient survival (p=0.0037 and p=0.049). Also large Ki 67/MIB1 positive surface (cut-off, 20%) correlated with disease-free survival in all patients and node positive patients (p=0.022 and p=0.0057) but not in node negative patients. It is concluded that in optimal technical conditions (automated and quantitative immunohistochemical assays on frozen sections) Ki 67 antigen immunohistochemical expression in breast carcinomas tissue has a prognostic significance in node positive patients but not in node negative patients.     

三阴性乳腺癌组织Ki-67指数预后价值分析

目的 Ki-67是细胞增殖的相关抗原,Ki-67指数是区分乳腺癌Luminal A型和Luminal B型的重要生物学指标,高Ki-67指数往往预示着不良的预后.然而在三阴性乳腺癌(triple negative breast cancer,TNBC)中,Ki-67预后价值尚不明确.本研究旨在探讨TNBC中Ki-67指数的预后价值.方法 回顾性分析郑州大学附属肿瘤医院2009-01-06-2010-12-30收治的310例经病理确诊为TNBC并有完整资料和随访数据患者的临床及病理资料,分析Ki-67指数等指标对患者生存预后影响.利用SPSS 17.0软件,计数资料比较采用χ2检验.Ki-67诊断价值及截断值采用ROC曲线进行分析.生存分析采用Kaplan-Meier法,并进行Log-rank检验.多因素分析采用Cox比例风险模型.结果 中位随访时间65个月(3~81个月),310例乳腺癌患者中复发68例(21.9%),死亡49例(15.8%),其中48例死于乳腺癌(15.5%).Ki-67指数与患者月经状态(χ2=8.484,P=0.014)、肿瘤大小(χ2=17.580,P=0.007)、腋窝淋巴结状态(χ2=30.071,P<0.001)以及组织学分级(χ2=17.626,P=0.001)均相关.低(Ki-67≤20%)、中(20%50%)5年无病生存率(disease-free survival,DFS)分别为96.5%、87.3%和64.9%,差异有统计学意义,P<0.001;5年总生存率(overall survival,OS)分别为96.5%、90.2%和75.5%,差异有统计学意义,P<0.001.Ki-67评价TNBC患者DFS及OS的ROC曲线下面积分别为0.707和0.689,Ki-67评价预后最佳截断值为57.5%.单因素分析中,Ki-67指数(χ2=31.779,P<0.001)、肿瘤大小(χ2=140.260,P<0.001)、腋窝淋巴结状态(χ2=120.467,P<0.001)和组织学分级(χ2=8.765,P=0.012)是影响TNBC患者DFS的相关因素,Ki-67指数(χ2=18.218,P<0.001)、肿瘤大小(χ2=299.718,P<0.001)、腋窝淋巴结状态(χ2=68.794,P<0.001)和组织学分级(χ2=7.572,P=0.023)是影响TNBC患者OS的相关因素;多因素分析中,Ki-67指数(HR=2.074,95%CI:1.279~3.364,P=0.003)、肿瘤大小(RR=1.879,95%CI:1.152~3.062,P=0.011)和腋窝淋巴结状态(RR=2.345,95%CI:1.825~3.015,P<0.001)是影响患者DFS的独立因素,Ki-67指数(RR=1.752,95%CI:1.020~3.008,P=0.042)、肿瘤大小(RR=20.011,95%CI:1.132~3.574,P=0.017)和腋窝淋巴结状态(RR=2.021,95%CI:1.517~2.693,P<0.001)是影响患者OS的独立因素.结论 Ki-67指数与TNBC患者预后相关,高Ki-67指数患者预后不良,Ki-67指数有望成为判断TNBC患者预后的一项重要生物学指标.     

Long term prognostic value of growth fraction determination by Ki-67 immunostaining in primary operable breast cancer

Summary An immunohistochemical determination of the growth fraction (GF) with the Ki-67 monoclonal antibody has been performed in a prospective series of 140 patients with primary operable breast carcinoma. GF ranged from 0% to 43% Ki-67 stained cells with a median value of 8%. High GF (> 8%) was significantly associated with axillary node involvement (p = 0.006), aneuploidy (p = 0.008), histologic grade (p = 0.03), and S-phase fraction > 5% determined by flow cytometry (p = 0.01). After a median follow-up of 6 years, the univariate analysis did not show significant correlation between high GF and worse relapse-free survival (p = 0.10) or shorter overall survival. However, a multivariate analysis on relapse-free survival, performed in 127 comparable patients, showed that GF was an independent predictive factor (p = 0.03) together with nodal status (p = 0.00001), age under 45 years (p = 0.0008), and chemotherapy (0.006). In node negative patients, GF was still an independent prognostic indicator (p = 0.002) together with age under 45 years (p = 0.0003). Tumor proliferative activity evaluated by the monoclonal antibody Ki-67 appears to be an effective indicator of prognosis in breast cancer and could be of assistance in the decision making of adjuvant therapy in node negative patients.     

Prognostic value of Ki-67 according to age in patients with triple-negative breast cancer

Purpose

The prognostic value of Ki-67 in triple-negative breast cancer (TNBC) is yet unclear because the cut-off points employed differ widely and its predictive effect may vary according to age. The purpose of this study was to analyze the role of Ki-67 among patients with TNBC, and determine the optimal Ki-67 cut-off point to demonstrate its prognostic relevance associated with patient age and treatment strategy.

Methods/patients

201 consecutive patients treated for primary TNBC from 1999 to 2014 were analyzed. Clinicopathological characteristics and outcomes were compared between patients treated with neoadjuvant or adjuvant chemotherapy. We used time-dependent receiver operating characteristic (ROC) curve and time-dependent area under the ROC curve (AUC) to evaluate the discriminative ability of Ki-67 at 3 and 5 years of follow-up. A Ki-67 cut-off point that maximized sensibility and specificity was established. Interaction effect between age and Ki-67 on disease-free survival (DFS) and overall survival (OS) was evaluated by stratified analysis.

Results

According to the coordinates of the ROC curves, the best cut-off point for Ki-67 was 60% (high/low). In the whole group, there was not a statistically significant association between Ki-67 and OS and DFS, using a cut-off point of 60%. In multivariate analysis (COX proportional hazards regression), for DFS high Ki-67 (>?60%) was a poor prognostic factor in patients?>?40 years old and a better prognostic factor among the patients?<?40 years old.

Conclusion

Prognostic value of Ki-67 in TNBC, using a cut-off point of 60%, may vary depending on age.

     

Prognostic value of Ki-67 labeling index in patients with node-negative, triple-negative breast cancer

The aim of this analysis was to investigate the usefulness of Ki-67 labeling index (LI) for the identification of different prognostic subgroups in primary node-negative, triple negative breast cancer (TNBC) patients. From January 1997 to December 2005, 1,053 patients operated for TNBC were identified through the institutional clinical database. The study was performed in accordance with REMARK criteria. The relationship between Ki-67LI and the risk of breast-related deaths was evaluated with a multivariable Cox regression model. Cubic splines were used to model Ki-67LI as a continuous variable. We selected 496 consecutive patients with node-negative TNBC. Median age was 52 years, median Ki-67LI 48% (range 4-95), and median follow up 6 years (range 0.5-13). Total deaths and deaths from BC were 52 (10.5%) and 38 (7.7%), respectively. Ki-67LI increased with decreasing age (P<0.01), increasing tumor size (P<0.01), and grade (P<0.01). When analyzing Ki-67LI as a continuous variable, the risk of death from BC increased steeply with increasing Ki-67LI up to about 35% and remained flat for higher values (adjusted effect of Ki-67 P=0.049; adjusted nonlinear effect P=0.021). Accordingly, when dividing patients into lower (≤35%) and higher (>35%) Ki-67LI subgroups, the 5-year cumulative incidence of breast-related deaths were 2.3 and 9.0%, respectively, with an adjusted HR(>35 vs ≤35) of 2.3 (95% CI 1.0-5.8, P=0.046). Within the group of patients with node-negative TNBC, Ki-67LI was associated with different prognoses subgroups. Ki-67LI might be useful in the design of trials of risk-adapted adjuvant therapies.     

Prognostic factors in operable breast cancer

A series of 743 consecutive cases of operable breast cancer, admitted and treated at the Istituto Nazionale Tumori of Milan from 1969 to 1970, was analyzed by a multivariate statistical method to evaluate a) the variables of the host and the primary tumor associated with the frequency of nodal metastases, b) the variables that significantly affect survival, and c) the identification of homogeneous risk groups. As regards the frequency of regional node metastases, they were more frequently observed in young than in old patients with large tumors (P values 10(-5) and 3 X 10(-5), respectively). Tumors that originated in the axillary tail, upper, outer and central quadrants were significantly associated with a higher rate of node metastases (P = 0.002). Each of these variables maintained its significant value when adjusted by the other two. Survival was affected at a statistically significant level by the age of the patients (P = 2 X 10(-4) ), the pathologic diameter of the primary tumor (P less than 10(-6) ), and the number of metastatic regional nodes (P less than 10(-6) ). The number of involved nodes appears to be the most relevant factor in the assessment of prognosis of patients with positive nodes, Age of the patients, size of the primary tumor, and number of involved nodes maintain their own statistical significance when each is adjusted by the remaining two. The site of origin of the primary tumor, even if associated with the frequency of regional node metastases, did not affect survival. Three groups with a significantly different risk of death were identified in patients with negative lymph nodes and three groups in patients with positive nodes. It is concluded that age, size of the primary, and number of involved lymph nodes are important pieces of information that clinicians should have at hand following radical surgery, not only to make a prognosis, but also to identify groups of patients with high risk of death on which the role of adjuvant treatment should be evaluated.     

Prognostic value of estrogen receptors in primary breast cancer.

The estrogen receptor status in 335 primary breast carcinomas was correlated with disease-free interval, survival and site of recurrent disease. Estrogen receptor positive carcinomas had a longer disease-free interval, a longer survival (mastectomy-death) and a longer time interval between recurrence and death. These parameters were also influenced by the lymph node status at mastectomy. Estrogen receptor positive cancers had a significantly better chance of survival independent of lymph node status. Estrogen receptors also delayed recurrence in node-positive carcinomas, but this advantage gradually disappeared with increasing interval after mastectomy. Estrogen receptor positive or estrogen receptor negative primary carcinomas did not show any predilection for spread to any particular site.     

Ki-67表达水平检测在乳腺癌新辅助化疗疗效评估中的价值

目的:回顾性分析88例乳腺癌新辅助化疗前、后Ki-67在肿瘤组织的表达情况,探讨Ki-67表达与新辅助化疗疗效的关系,评价其在乳腺癌新辅助化疗中的预测作用.方法:选取2015年9月至2016年9月河北医科大学第四医院乳腺中心收治的88例Ⅱ-Ⅲ期乳腺癌患者,检测新辅助化疗前空芯针穿刺肿瘤组织及术后标本中Ki-67的表达,分析其与新辅助化疗疗效及临床相关病理因素的关系.结果:新辅助化疗的临床总有效率为59.09%(52/88),Ki-67高表达组对化疗敏感,化疗效果明显优于Ki-67低表达组(P<0.05);新辅助化疗可明显降低Ki-67的高表达率(P<0.01);新辅助化疗后Ki-67表达下降组化疗有效率显著高于其他组(P<0.05).结论:Ki-67在乳腺肿瘤组织中的表达可作为新辅助化疗疗效临床评价指标之一,预测新辅助化疗的疗效,为个体化治疗提供依据.     

Proliferation marker Ki-67 in early breast cancer.

Molecular markers have been extensively investigated with a view to providing early and accurate information on long-term outcome and prediction of response to treatment of early breast cancer. Proliferation is a key feature of the progression of tumors and is now widely estimated by the immunohistochemical assessment of the nuclear antigen Ki-67. The expression of Ki-67 correlates with other measurements of proliferation, including S-phase and bromodeoxyuridine uptake. High Ki-67 is a sign of poor prognosis associated with a good chance of clinical response to chemotherapy, but its independent significance is modest and does not merit measurements in most routine clinical scenarios. However, its application as a pharmacodynamic intermediate marker of the effectiveness of medical therapy holds great promise for rapid evaluation of new drugs.     

Ki-67 labeling index in breast cancer

Measurements of cell cycle kinetics have been found to correlate with the clinical course of patients with breast cancer. However, the thymidine labeling index and more rapid methods like flow cytometry remain complicated and costly. We assessed cell proliferation of 67 breast carcinomas by an immunoperoxidase procedure using a monoclonal antibody, Ki-67, which reacts with a nuclear antigen in proliferating cells. The percentage of Ki-67 positive cells ranged from 2% to 70%. Tumors with high mitotic rate, high nuclear grade, high histologic grade, and negative estrogen receptors had statistically higher Ki-67 labeling rates. We found no significant differences between the Ki-67 labeling rate and other clinical (age at diagnosis, menopausal status) or pathologic (necrosis, fibrosis, vascular invasion, lymphatic invasion, cellular reaction, tumor size, lymph node metastases) features assessed. These results parallel previously reported data, and confirm that this immunohistochemical staining of breast carcinoma by Ki-67 monoclonal antibody can be considered a rapid and convenient method for assessing cell cycle kinetics. However, further studies, evaluating the correlation between Ki-67 labeling rate and prognosis are needed to better define the real usefulness of this analysis in clinical practice.     

Prognostic value of Ki67 expression after short-term presurgical endocrine therapy for primary breast cancer

Tumor expression of the proliferation antigen Ki67 is widely used to assess the prognosis of cancer patients. A change in the expression of Ki67 after short-term exposure of patients to therapeutic agents is frequently used as a pharmacodynamic marker of efficacy, particularly among breast cancer patients before undergoing surgery. To determine the clinical significance of the level of tumor cell proliferation during endocrine therapy for breast cancer, we measured the expression of Ki67 in tumor biopsy samples taken before and after 2 weeks of presurgical treatment with anastrozole or tamoxifen or the combination of anastrozole plus tamoxifen in 158 patients with hormone receptor-positive primary disease. In a multivariable analysis, we found that higher Ki67 expression after 2 weeks of endocrine therapy was statistically significantly associated with lower recurrence-free survival (P = .004) whereas higher Ki67 expression at baseline was not. Larger baseline tumor size and lower estrogen receptor level after 2 weeks of treatment were also statistically significantly associated with poorer recurrence-free survival (P < .001 and P = .04, respectively). Our data indicate that measurements of tumor Ki67 level after short-term endocrine treatment may improve the prediction of recurrence-free survival by integrating the prognostic value of Ki67 level at baseline with changes in Ki67 level that are associated with treatment benefit.     

Prognostic value of residual node involvement in operable breast cancer after induction chemotherapy

The purpose of this retrospective study was to evaluate the influence of axillary disease on patients' survival after neoadjuvant chemotherapy and to assess patient and tumor characteristics associated with post-chemotherapy axillary involvement.After six induction cycles, 277 patients with operable breast cancer (stage II–III) underwent surgery with axillary dissection, followed by radiotherapy (n = 267) or additional chemotherapy (n = 63) and adjuvant tamoxifen therapy (n = 138). At a median follow-up of 8.5 years, overall survival (OS) and disease-free survival (DFS) were analyzed as a function of node involvement.The differences in OS and DFS according to the number of positive nodes were highly statistically significant with a decreased survival associated with the increasing number of nodes (p = 5 × 10^–6 and 9 × 10^–7, respectively). Upon multivariate analysis, the node number after chemotherapy appeared as the most significant prognostic factor (p = 7 × 10^–4 for OS and p = 3 × 10^–5 for DFS). All the other classical prognostic factors were insignificant, except post-chemotherapy Scarff–Bloom–Richardson (SBR) grading for OS (p = 8 × 10^–4) and adjuvant hormonotherapy for DFS (p = 1 × 10^–2).Although constituting a different parameter from primary surgery data, the number of positive nodes after chemotherapy could still remain a valuable prognostic factor at secondary surgery, raising the question for high risk patients of a second non-cross-resistant adjuvant regimen, or high dose chemotherapy with peripheral blood stem cells support.     

肾癌细胞Ki-67免疫组化染色指数与预后的关系

 为探索肾癌预后较好的估价指标，41例肾癌根治标本Ki—67免疫组化染色。复发组的Ki—67染色指数显著高于未复发组（P相似文献   

Predictive value of biologic parameters for primary chemotherapy in operable breast cancer

Primary chemotherapy represents an ideal model to evaluate the relationships between treatments and the prognostic and predictive parameters provided by the new technologies. First- and second-generation trials have shown that primary chemotherapy significantly improves the rate of breast conservation without increasing the risk of ipsilateral recurrence and while assuring survival rates comparable with those achieved with postoperative chemotherapy. Moreover, patients who exhibited a pathologic complete response (pCR) showed better progression-free survival and overall survival. The third-generation trials were aimed at improving the percentage of pCR, identifying and validating gene and protein biomarkers of chemotherapy sensitivity, and better defining the individual risk of relapse. Several parameters, such as index of proliferation and apoptosis, expression of proteins (eg, p53 and Bcl-2), and hormone receptor and epidermal growth factor family receptors, have been related to response to primary chemotherapy. Negative hormone receptors and greater proliferative activity seem to be the only parameters more consistently associated with greater chemotherapy sensitivity. However, the strength of this association is not sufficient to differentiate patients at different degrees of risk and does not allow for an individualized therapeutic choice. Newer technologies offer the possibility of evaluating thousands of genes and identifying clusters of gene expression associated with significantly different risks of relapse and patterns of sensitivity/resistance to specific drugs. The primary chemotherapy model is the ideal clinical setting in which to validate the relationship between tumor molecular profiling and treatment outcomes and to design tailored therapies based on observed effects on individual tumors.     

Prognostic and predictive indicators in operable breast cancer

Because of its biological heterogeneity and wide spectrum of responsiveness to different treatments, breast cancer is a complex disease of difficult clinical management. Over the past several years, knowledge of the molecular mechanisms regulating normal and aberrant cell growth leading to cancer has been enhanced. These advances have enabled the identification of an increasing number of surrogate biomarkers, which have been correlated with prognosis or used as predictors of response to specific treatments. Axillary nodal status, age, tumor size, pathologic grade, and hormone receptor status are the established prognostic and/or predictive factors for selection of adjuvant treatments. The role of new biomarkers, such as p53, HER2/neu, angiogenesis, and the proliferation index value, is promising; however, the clinical value of their determination must be provided by prospective clinical studies.     

Comparison of visual and automated assessment of Ki-67 proliferative activity and their impact on outcome in primary operable invasive ductal breast cancer

Background:

Immunohistochemistry of Ki-67 protein is widely used to assess tumour proliferation, and is an established prognostic factor in breast cancer. There is interest in automating the assessment of Ki-67 labelling index (LI) with possible benefits in handling increased workload, with improved accuracy and precision.

Patients and methods:

Visual and automated assessment of Ki-67 LI and survival were examined in patients with primary operable invasive ductal breast cancer. Tissue microarrays (n=379 patients) immunostained for Ki-67 were scored visually and automatically with the Slidepath Tissue IA system.

Results:

Visual and automated Ki-67 LI were in excellent agreement (ICCC=0.96, P<0.001). On univariate analysis, visual (P<0.001) and automated Ki67 LI (P<0.05) were associated with cancer-specific survival in patients with invasive ductal breast cancer overall and in patients who received endocrine therapy (Tamoxifen) (P<0.01 for visual and P<0.05 for automated scoring).

Conclusion:

Automated assessment of Ki-67 LI would appear to be comparable to visual Ki-67 LI. However, automated Ki-67 LI assessment was inferior in predicting cancer survival in patients with breast cancer, including patients who received Tamoxifen.     

Ki-67在早期乳腺癌诊治中的意义

早期乳腺癌(EBC)的预后差异非常大,重要的预后因素有肿瘤大小、组织学分级、脉管浸润情况、淋巴结转移情况等是有所帮助的,肿瘤的分子特征也正受到越来越多的关注,ER、Her-2已经被作为治疗反应和预后的预测指标.Ki-67虽然不作为必须的标记,但常用来作为增殖活性的静态标志,或通过治疗期间的多次测量,来反映动态变化的中间状态,以及作为疗效的替代指标.现应就Ki-67在早期乳腺癌的预后和治疗的预测价值作一综述.     

Prognostic value of CD44 variant expression in primary breast cancer.

CD44 is a family of cell surface transmembrane glycoproteins members which differ in the extracellular part by sequences derived by alternative splicing of 10 variant exons (v1-v10). CD44 proteins containing such variant sequences have been implicated in tumor metastasis formation. Here, we have evaluated the expression of CD44 variants by immuno-histochemistry in primary breast cancer samples of 237 node-negative and 230 node-positive patients. For the analysis of samples derived from node-negative patients, the exon-specific antibodies used were DIII, vff7 and vff18 (v6), vff17 (v7/v8), fw11.24 (v9) and vff16 (v10). With the different antibodies which recognize v6 epitopes, the majority of tumors were positively stained (> or = 65% of the tumors) with varying intensities. Thirty-nine percent of the tumors were positively stained with the antibody vff16, and approximately half of the tumors with the antibodies vff17 and fw11.24. The expression of CD44 v6 epitopes in tumors from node-negative patients was associated with a favorable prognosis, both upon univariate and multivariate analysis. The expression of CD44 v7/8, v9 or v10 epitopes was not significantly related with relapse-free survival. Samples from node-positive patients were only examined with the antibodies vff7, vff17 and vff18. The staining with none of these antibodies was correlated with the length of relapse-free survival of the patients. Our data suggest that, generally, the usefulness of knowledge of CD44 variant expression is of limited value for assessing the risk of relapse in patients with primary breast cancer. However, the expression of exon v6 of CD44 may be a marker to identify patients with a relatively favorable prognosis in node-negative patients.     

Prognostic impact of proliferation-associated factors MIB1 (Ki-67) and S-phase in node-negative breast cancer.

MIB1 proliferation rate (MIB1-PR) and total S-phase fraction (SPF) were retrospectively determined in formalin-fixed, paraffin-embedded sections of 90 primary node-negative breast carcinomas. None of the patients had received adjuvant systemic therapy. Median follow-up in patients still alive at the time of analysis was 37.5 months (16-72 months). Immunostaining of Ki-67 antigen was performed using the monoclonal antibody MIB1 and the APAAP technique. An adjacent 50-microm paraffin section was used for flow cytometric S-phase determination. Results were compared to established clinicopathological prognostic factors. MIB1-PR was significantly correlated to grading (P = 0.018); SPF was significantly correlated with tumour size (P = 0.041) and inversely with steroid hormone receptor status (P = 0.03). A significant correlation between MIB1-PR and SPF was found in aneuploid (P = 0.025) but not in diploid tumours (P = 0.164). In univariate analysis, both MIB1-PR (optimized cut-off of 25%) and SPF (optimized cut-off of 8%) were significant prognostic factors for disease-free survival (DFS) (MIB1-PR, P = 0.0224; SPF, P = 0.0028). In multivariate analysis, however, only SPF remained significant; it was the strongest prognostic factor for DFS (P = 0.0073), stronger than MIB1-PR or established clinicopathological prognostic factors. We thus conclude that MIB1-PR and SPF provide additional prognostic information in node-negative breast cancer. However, in our study, flow cytometrically determined SPF had the greater prognostic impact.