Prognostic significance of Ki-67, p53, and Bcl-2 expression in prostate cancer patients with lymph node metastases: A retrospective immunohistochemical analysis

The prognostic significance of Ki-67, p53, and Bcl-2 expression was evaluated in prostate cancer patients with lymph node metastases. Immunohistochemical staining of archived material obtained from 56 patients was performed by the streptavldin-biotin method. Univariate survival analysis showed that a Ki-67 labeling index (Ki-67 LI) of ≥8.4 in the primary tumor identified a group of patients with a significantly poorer prognosis (P<0.001). Furthermore, a Ki-67 LI of ≥8.7 in the nodal metastatlc tumor was also associated with a poorer prognosis (P<0.01). Multivariate analysis showed that the Ki-67 LI of primary tumors (P<0.01) and lymph node metastases (P<0.01) had independent prognostic value. p53 and Bcl-2 expression had no prognostic value in patients with prostate cancer and lymph node involvement. The Ki-67 LI has more prognostic value than p53 and Bcl-2 expression for patients with prostate cancer that has spread to the lymph nodes.     

Nucleolar organizer regions relate to growth fractions in human breast carcinoma

An argyrophil stain for nucleolar organizer regions (NORs) has recently been applied to paraffin sections of human tissues. This report describes a positive relationship between the mean numbers of AgNOR sites per nucleus and tumor growth fraction, as determined by immunostaining with the monoclonal antibody Ki-67, in 83 malignant breast tumors (P less than .01). This relationship supports recent suggestions that the NOR count may reflect cell synthetic activity and hence, proliferation. AgNOR counts correlated inversely with immunocytochemically assessed estrogen receptor content (P less than .002), but there was no relationship between the AgNOR count and primary tumor size, histologic grade, axillary node status, or patient age. A significant difference (P less than .00001) was found between the AgNOR counts in 64 benign breast lesions (mean, 2.05) and 85 malignant breast neoplasms (mean, 5.46). The limitations of the silver staining technique and the problems of reproducibility in AgNOR counting are detailed.     

Prognostic significance of p34cdc2 cyclin-dependent kinase and MIB1 overexpression, and HER-2/neu gene amplification detected by fluorescence in situ hybridization in breast cancer.

The HER-2/neu oncogene, localized to chromosome 17q, shares substantial homology with the epidermal growth factor receptor. HER-2/neu gene amplification and protein overexpression have been associated with poor prognosis in breast cancer. Formalin-fixed paraffin-embedded primary invasive breast cancer tissues from 135 women were tested for HER-2/neu gene amplification by automated fluorescence in situ hybridization (FISH) using a sequence probe. The tumors also were evaluated by immunohistochemistry for proliferation markers Ki 67 (MIB1) and p34cdc2 cyclin-dependent kinase. Patients were followed up for a mean of 61 months. There were 70 node-negative and 65 node-positive cases. Ki 67 and p34cdc2 proliferation marker overexpression, HER-2/neu oncogene amplification, large tumor size, high tumor grade, advanced tumor stage, positive lymph node status, and distant metastasis at the time of diagnosis predicted disease-related death in combined node-negative and node-positive breast cancer. HER-2/neu gene amplification, tumor stage, lymph node metastasis, tumor grade, and distant metastasis at the time of diagnosis independently predicted disease outcome. HER-2/neu amplification detected by FISH also predicted disease-related death independent of lymph node status, tumor grade, and distant metastasis in breast cancer patients who received adjuvant therapy.     

Detection of Ki-67 proliferation rate in breast cancer. Correlation with clinical and pathologic features

In situ determination of proliferative activity was performed on 203 breast cancers by use of Ki-67 monoclonal antibody and immunohistochemical methods. Tumor proliferation rate was analyzed and correlated to tumor size and nodal status. The relationship between Ki-67 proliferative activity and nuclear estrogen receptor content was also investigated on adjacent tissue sections. Ki-67 values ranged from 1 to 75%, with a median value of 10%. Premenopausal patients had greater Ki-67 values (median value, 14.1%) than postmenopausal ones (median value, 9.8%). The authors observed no correlation with lymph nodal involvement, whereas a statistically significant relationship with tumor size was found (P less than 0.01). An inverse correlation (Spearman's coefficient = -0.56; P less than 0.001) was seen between Ki-67 values and nuclear estrogen receptor content. These results, similar to those reported for other kinetic measurements, suggest that in situ detection of Ki-67 proliferation rate is a useful method for obtaining cell cycle information. Follow-up studies will be needed to assess an eventual prognostic relevance.     

Mucin antigens expression and Ki-67 labeling in breast cancer: The peculiarity in scirrhous carcinoma

The expression of mucin-related antigens (Tn, T, Sialosyl-Tn [STn], DF3 [mammary-type apomucin related antigen], and intestinal-MRP [intestinal-type apomucin related antigen]) as well as Ki-67 labeling was examined in 58 mammary invasive ductal carcinomas (IDC) divided into 26 scirrhous subtype (SC) and 32 non-scirrhous subtype comprising papillotubular carcinoma and solid-tubular carcinoma (PT-ST). These data were analyzed in connection with the various pathological prognostic factors such as nodal status, tumor size, estrogen receptor status and histological grading of carcinomas. The results were as follows: (a) in SC, the expression rate of Tn was significantly higher in the cases with positive lymph node metastasis or with large tumor size (>2cm) than in those with negative lymph node metastasis or with small tumor size (>2 cm); (b) in PT-ST, the expression rate of STn was higher in the cases with positive lymph node metastasis or with large tumor size than in those with negative lymph node metastasis or with small tumor size; (c) in SC, Ki-67 labeling was significantly higher in the cases with positive lymph node metastasis than in those with negative lymph node metastasis; and (d) in PT-ST, Ki-67 labeling was lower in the cases with positive lymph node metastasis than in those with negative lymph node metastasis. In conclusion, Tn antigen expression was correlated with pathological prognostic factors in SC but not in PT-ST, whereas STn antigen expression was correlated with pathological prognostic factors in PT-ST but not in SC. Moreover, lnverse relationship between Ki-67 labeling and nodal status was observed in PT-ST. These differences between SC and PT-ST may be related to their different biological behaviors.     

The morphometric prognostic index is the strongest prognosticator in premenopausal lymph node-negative and lymph node-positive breast cancer patients

Earlier studies on breast cancer have shown the strong prognostic value of morphometric parameters (especially the morphometric prognostic index [MPI]) in comparison with clinical and classical pathologic parameters. It remained to be proven whether the prognostic value of the MPI holds for the subgroup of premenopausal patients. We have therefore investigated the value of different prognosticators in a group of 211 premenopausal breast cancer patients with long-term follow-up, 121 cases being lymph node-negative and 90 cases being lymph node-positive. The MPI, a multivariate combination of the mitotic activity index (MAI), lymph node status, and tumor size, was the best combined prognosticator (P less than .0001), exceeding the prognostic value of MAI, lymph node status, and tumor size as individual parameters and as indicators of histologic grade. Of all the features studied, the MPI had the best prognostic value in the lymph node-negative patients, while the MAI and MPI had the best prognostic value in the lymph node-positive patients. Since the MPI has been shown to be reproducible in intra- and interlaboratory studies and can be assessed with standard equipment in routine histologic sections, it is an attractive indicator for selecting high-risk lymph node-negative patients for systemic adjuvant therapy trials.     

白细胞介素-1β的表达及间质微血管密度在乳腺浸润性微乳头状癌中的意义

目的 研究白细胞介素(IL)-1β的表达和间质微血管密度在乳腺浸润性微乳头状癌(IMPC)中的意义.方法 采用免疫组织化学LSAB法检测100例IMPC、97例浸润性导管癌(IDC)中IL-1β和CD34的表达并计数微血管密度,比较其差异并分析IMPC中IL-1β、CD34的表达与ER、PR、肿瘤细胞增殖指数Ki-67、组织学分级和淋巴结转移等主要病理学特征的关系.结果 (1)IMPC中IL-1β的表达与IDC差异无统计学意义(P=0.924);(2)IMPC中IL-1β的表达与Ki-67的表达(x2=7.538,P=0.023)、组织学分级(x2=6.556,P=0.038)及淋巴结转移个数(P=0.008)呈正相关,而与ER呈负相关(z=-2.106,P=0.035);(3)微血管密度:IMPC组(66.4±15.9)明显高于IDC组(60.0±14.1,t=2.995,P=0.003),且在IMPC中淋巴结转移组(68.8±13.9)显著高于无淋巴结转移组(54.4±20.7,t=-3.459,P=0.001);IMPC中组织学Ⅱ、Ⅲ级组(68.3±15.0)显著高于I级组(59.9±17.6,t=-2.281,P=0.025),而微血管密度与ER、PR、Ki-67表达无相关性.结论 IL-1β表达增高、间质微血管密度增加可能是促进乳腺IMPC肿瘤细胞增殖和淋巴结转移的关键因素.     

Computer-assisted analysis of cell proliferation markers in oral lesions

Abnormalities in any component of the cell cycle regulatory machine may result in oral cancer, and markers of cell proliferation have been used to determine the prognosis of tumor progression. The aim of this study was to determine whether silver-stained nucleolar organizer region (AgNOR) and Ki-67 measurements could improve the assessment of growth rates in oral lesions. Eighty-three oral biopsies were studied, 20 of which were classified as fibrous inflammatory hyperplasia (FIH), 40 as leukoplakia (LKP) and 23 as oral squamous cell carcinoma (OSCC). Within the LKP group, 22 out of 29 biopsies were diagnosed as non-dysplastic leukoplakia (LK) and 18 as dysplastic leukoplakia (DLK), presenting discrete, moderate and severe dysplasia. Ki-67 immunolabeling of the lesions increased steadily in the following order: FIH, DLK, LK and OSCC, indicating that Ki-67 is a good marker for predicting the proliferative fraction among benign, premalignant and malignant oral lesions. The median values of AgNOR parameters indicate that the morphometric index gives better results regarding the proliferative rate than the numerical one. A series of linear regressions between AgNOR parameters and Ki-67 showed positive associations. We conclude that a combination of Ki-67 and morphometric AgNOR analyses could be used as an aid in the determination of the proliferative status of oral epithelial cells in oral cancer.     

Comparison of proliferative activities and metastases between two subtypes classified at the deeply infiltrating sites of colorectal moderately differentiated adenocarcinomas

Twenty-eight moderately differentiated adenocarcinomas invading beyond the muscularis propria of the colorectum were subclassified as 13 moderate- and 15 poor-subtype tumors based on the histology at the deeply infiltrating sites. Moderately differentiated cancer cells were correlated with liver metastasis and p53 immunoreactivity. Poorly differentiated cancer cells were correlated with lymph node metastases but not to p53 immunoreactivity. The proliferative cell nuclear antigen (PCNA) labeling index (LI), Ki-67 LI and agyrophilic nuclear organizer regions (AgNOR) values determined for the poorly differentiated cancer cells in the poor-subtype tumors were significantly lower than those of moderately differentiated cancer cells in the moderate-subtype tumors. In cells from tumors classified as poor-subtype, poor differentiation was associated with decreased PCNA LI levels, but with unchanged Ki-67 LI and AgNOR values. These results indicate that colorectal adenocarcinoma cells that are histologically subclassified as moderately differentiated have different proliferative and metastatic activities from cancer cells that are poorly differentiated. Moderately differentiated cancer cells are associated with hematogenous metastasis to liver and high proliferative activity, and loss of tubular formation of cancer cells may be fundamentally related to lymph node metastasis and infiltrative growth.     

Silver-stained Nucleolar Organizer Regions (AgNOR)]

The present paper discusses the relevance of silver-stained Nucleolar Organizer Regions (AgNOR) to tumour pathology. First, the structural and functional aspects of AgNORs and the biological meaning of their quantitative variations have been reviewed to clearly define which indications can be obtained by the use of AgNOR. In continuously proliferating cells, AgNOR indicates the extent of ribosomal biogenesis, which is strictly related to the rapidity of cell proliferation. Therefore, AgNOR can be considered to represent a marker of cell proliferation rate and, in tumour pathology, should be used only for prognostic purposes. The predictive value of AgNOR, the only parameter which indicates the cell growth rate in situ in routinely processed cyto-histological samples, is strengthened by the combination with markers of cell growth kinetics (e.g. Ki67/MIB1).     

The prognostic importance of the nucleolar organizer region (AgNOR), Ki-67 and proliferating cell nuclear antigen (PCNA) in primary nonurachal bladder adenocarcinoma

The aim of this study was to investigate the role of tumor proliferation in patients with nonurachal bladder adenocarcinoma. Samples were obtained from 16 patients (12 men and 4 women, mean age 62 years) with primary nonrurachal bladder adenocarcinoma. The 16 formalin-fixed specimens were stained immunohistochemically for Ki-67 antigen and PCNA using MIB-1 and PC-10 antibodies. In addition, the AgNOR quantity was assessed using the colloid silver nitrate staining technique in all cases. The Ki-67, PCNA and AgNOR proliferation indices were found to be significantly higher in high-grade and invasive tumors. The higher the grade (p<0.01) and stage (p<0.01), the higher were the proliferation indices. Patients whose tumor samples had a high Ki-67, PCNA and AgNOR proliferation index showed a higher incidence of local recurrence (p<0.01) and distant metastasis (p<0.01). In conclusion, our results suggest that Ki-67, PCNA and AgNOR proliferation scores may be important prognostic indices in nonurachal bladder adenocarcinomas.     

Lymphangiogenesis in Breast Cancer Correlates with Matrix Stiffness on Shear-Wave Elastography

PurposeTo correlate tumor stiffness and lymphangiogenesis in breast cancer and to find its clinical implications.ResultsHigher elasticity value was associated with invasive size of tumor, microlymphatic density, histologic grade 3, absence of extensive intraductal component, presence of axillary lymph node metastasis, and Ki-67 labeling index (LI) in univariate regression analysis, and associated with Ki-67 LI and axillary lymph node metastasis in multivariate regression analysis. Microlymphatic density was associated histologic grade 3, mean elasticity value, and Ki-67 LI in univariate regression analysis. In multivariate regression analysis, microlymphatic density was correlated with mean elasticity value.ConclusionIn breast cancer, tumor stiffness correlates with lymphangiogenesis and poor prognostic factors.     

Proliferation in squamous cell carcinoma of cervix: a comparative assessment by two markers

The study was aimed at assessing the cell proliferation in various grades of squamous cell carcinoma of cervix using two proliferation markers: AgNORs and Ki-67. 18 biopsy sections of various grades of SCC of cervix were chosen. AgNORs staining was done by single step staining technique using gelatin. Ki-67 immunostaining was done by Avidin--Biotin technique using DAB as a chromogen. Highest mean number of AgNORs was found in poorly differentiated squamous cell carcinoma and statistically significant difference was observed between well and poorly differentiated SCC and between moderately and poorly differentiated SCC. Highest mean Ki-67 LI was seen in poorly differentiated SCC. We observed correlation between histological grade and Ki-67 and AgNORs separately but no statistically significant correlation was found between Ki-67 and AgNOR counts. AgNORs and Ki-67 are both simple and easily performed techniques. They may prove to be a useful adjunct in estimating tumor proliferation and hence in determining the management strategy of the patients.     

Correlation of nucleolar organizer regions and nuclear morphometry assessed by automatic image analysis in breast cancer with aneuploidy, K167 immunostaining, histopathologic grade and lymph node involvement.

Silver-stained nucleolar organizer regions (AgNORs) in human breast carcinoma were studied using a computer-assisted system of image analysis. Standardized, automatic measurements of 7 morphometric parameters (area, perimeter, shape factor, bend energy, angle, and small and large diameters) performed on paraffin sections and cell imprint were compared and correlated with nuclear morphometry, histopathological grading, tumor growth fraction, (monoclonal Ki67-immunostaining), DNA nuclear content (stoechiometric Feulgen staining) and axillary lymph node invasion. The major findings were as follows: (i) variations in AgNORs and nuclear parameters were correlated, (ii) the ratio of AgNOR area/nuclear area was significantly different in low and high grade tumors, (iii) mean AgNOR parameter values increased significantly with the tumor growth fraction and tumor hyperploidy and were significantly higher in patients with axillary lymph node metastases and (iv) AgNOR evaluation was more accurate for cell preparations than for tissue sections.     

Grading and prognosis of invasive ductal mammary carcinoma by nuclear image analysis in tissue sections

In primary tumours of 84 patients with invasive ductal carcinomas the prognostic significance of the nuclear image was assessed. Feulgen stained tissue sections were investigated by image cytometry. At least 150 tumour cell nuclei as well as 50 lymphocytes or trout erythrocytes for diploid reference were measured in each specimen. 35 morphological features, including those of the chromatin pattern, were derived from the digitized nuclear images. By means of multivariate discriminant analysis, significant differences could be found between tumours of 41 node-negative and 43 node-positive patients. The differences were sufficiently large to distinguish subgroups with low and high risk of lymph node involvement. Irrespective of lymph node status, 66 patients with an adequate 5 year follow-up were divided into two groups according to their survival time. The classification results revealed a clear relationship between characteristics of the nuclear image and prognosis. By image cytometry it is thus possible to separate both node-positive and node-negative patients into subgroups with increasing risk of recurrence. The results indicate that nuclear image analysis provides prognostic information in addition to established prognostic factors, such as histological grade and tumour stage.     

Correlation of two argyrophilic nucleolar organizer region counting methods with the Ki-67 labeling index in uterine smooth muscle cell tumors

The argyrophilic nucleolar organizer regions (AgNOR) are silver stained granules that are thought to correlate with cell proliferation activity. Two AgNOR counting methods: the mean AgNOR count (mAgNOR, the mean number of AgNOR granules in 100 cells) and the AgNOR proliferative index (pAgNOR, the percentage of cells exhibiting five or more AgNOR granules per nuclei) have been proposed. In this study, the two counting methods were applied to 58 cases of normal uterine corpus and uterine corpus tumors and were compared with the Ki-67 labeling index (Ki-67 LI) using MIB-1 monoclonal antibody and other histopathological criteria. Notable differences in the number of AgNOR and the Ki-67 LI were observed between benign and malignant smooth muscle tissue. Histopathologic features are well correlated to the proliferative activity of tumors. Although the most reliable method of predicting malignant potential cannot be determined, the methods outlined by this study are thought to be highly useful in assessing proliferative activities.     

细胞周期蛋白D1、Ki-67和bcl-2的表达与CD117阳性的胃肠道间质瘤生物学行为的关系

目的 检测细胞周期蛋白D1（cyclin D1）、Ki-67和bcl-2在胃肠道间质瘤（GIST）中的表达,探讨它们在GIST发生、发展中的作用及临床病理意义。方法 59例手术切除GIST标本进行CD117、CD34、平滑肌肌动蛋白（SMA）、结蛋白、S-100、cyclin D1、bcl-2和Ki-67免疫组织化学染色,同时进行病理形态学观察包括形态学类型、肿瘤大小、坏死和核分裂象。所有病例随访2～9年。所有数据进行单因素、多因素和相关分析。结果 随访40例患者一直健在,15例患者死于GIST,4例患者死于其他原因。统计学分析显示肿瘤直径〉5cm、有坏死、核分裂象在每50个高倍视野〉5个、Ki-67增殖指数（LI）〉5％、cyclin D1和bcl-2免疫组织化学染色强阳性都可以作GIST患者手术后的预测指标,且具有统计学意义;核分裂象和Ki-67增殖指数是独立的预测指标;Ki-67 LI≥5％和核分裂象≥5／50 HPF呈正相关（r=0．532,P〈0．01）;cyclin D1与bcl-2强阳性表达呈正相关（r=0.273,P〈0．05）。结论 肿瘤大小、坏死、核分裂象、cyclin D1、Ki-67增殖指数和bcl-2可作为GIST患者临床预测指标;核分裂象和Ki-67增殖指数可作为独立的预测指标;cyclinD1与bcl-2呈明显相关性,Ki-67免疫组织化学染色可以代替核分裂象作为一项很有用的预测指标。     

Clinical significance of cell proliferation, microvessel density, and CD44 adhesion molecule expression in renal cell carcinoma.

Renal cell carcinoma (RCC) is known to have a wide variation in clinical outcome despite the use of conventional prognostic factors, such as staging or grading. A better knowledge of the biologic aggressiveness of RCC could facilitate the selection of patients at high risk of tumor progression. The aim of this study was to determine if use of measurements of vascular density, cell proliferation, and cell adhesion could better predict the biologic behavior of RCC. We immunohistochemically analyzed CD34, Ki-67, and CD44H expression on formalin-fixed, paraffin-embedded tissues from 73 RCCs for quantifying microvessel density (MVD), Ki-67 labeling index (LI), and CD44H LI, respectively. Univariate cancer-specific survival analysis showed that tumor stage (P < .01), tumor size (P < .001), nuclear grade (P < .01), metastasis (P < .001), MVD (P < .03), Ki-67 LI (P < .001), and CD44H LI (P < .0001) were predictors of tumor-related death. There was a statistical correlation between CD44H LI and both Ki-67 LI (r' = .3) and MVD (r' = -44). Ki-67 LI (P < .04) and CD44H LI (P < .02), as well as metastasis (P < .008), emerged as independent predictors of cancer-specific survival in multivariate analysis in patients with metastases (P < .04 and P < .02, respectively) and in patients without metastases (P < .006 and P < .00001, respectively). Our study suggests that vascular density, cell proliferation, and cell adhesion represent a complex tumor-host interaction that may favor progression of RCC. Cell proliferation and CD44H expression appear to be powerful markers to identify patients with an adverse prognosis.     

Histopathological subclassification of triple negative breast cancer using prognostic scoring system: five variables as candidates

We attempted to subclassify triple negative breast cancer (TNBC) cases into subgroups according to clinical outcome or prognosis of TNBC patients using archival specimens. We analyzed 102 Japanese cases of invasive TNBC who underwent surgery between January 1998 and December 2007. The clinicopathological factors and clinical information were retrospectively retrieved from reviewing the charts of the patients. Immunohistochemical staining was performed for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), epidermal growth factor receptor 1 (EGFR1), CK5/6, CK14, Ki-67, and CD31 for microvessel density (MVD). Median follow-up time of the patients was 68.5 months. Multivariable analysis demonstrated that pathologic node status was the most significantly associated with relapse-free survival (RFS) and breast cancer-specific survival (BCSS) of these patients. Pathological tumor size, basal-like type, Ki-67 labeling index (LI) and MVD were also independently associated with RFS and BCSS. Based on these results, we devised the risk score system reflecting hazard ratios of these prognostic factors above. With this system, TNBC patients in this study were classified into three subgroups (low-risk group: score 0–3, intermediate-risk group: score 4–7 and high-risk group: score 8–10). The significant difference of RFS and BCSS was detected among these three different subgroups of the patients (p < 0.05). We propose the risk score system, which incorporated pathologic nodal status, size of the primary tumor, the presence or absence of basal-like features, Ki-67 LI, and MVD in order to predict postoperative clinical course of the Japanese TNBC patients.     

Proliferation characteristics in pediatric Hodgkin's lymphoma point to a cell cycle arrest in the G(1) phase.

This study was undertaken to determine the prognostic relevance of the proliferation rate in neoplastic cells in children and adolescents with Hodgkin's lymphoma. Paraffin-embedded biopsy specimens were immunostained with the proliferation-associated monoclonal antibodies Ki-S5 (Ki-67 antigen) and Ki-S2 (which detects the repp86 protein). Repp86 is a protein of about 100 kDa encoded by a gene located on human chromosome band 20q11.2. In contrast to the Ki-67 antigen, repp86 expression is restricted to the cell cycle phases G(2), S and M. Immunohistochemical results on diagnostic lymph node biopsy specimens from 224 patients included in two pediatric multicenter Hodgkin's trials, GPOH HD-90 and HD-95, were compared with clinical data. High Ki-67 antigen expression was a striking feature of Hodgkin's and Reed-Sternberg cells as well as lymphocytic and histiocytic cells (median: 80%, range: 20-100%), in contrast to low repp86 expression (median: 20%, range: 10-80%; P<0.001). The proliferation rate was independent of histological subtype, stage and presence of B symptoms. The probability of event-free and overall survival (+/-standard error) of all patients at 5 years was 91.6+/-2.0 and 98.1+/-1.0%, respectively. The proliferation rate of tumor cells did not influence the outcome. The difference between Ki-67 and repp86 expression in Hodgkin's and Reed-Sternberg or lymphocytic and histiocytic cells points to a possible cell cycle arrest in the G(1) phase, which may explain the obvious paradox of a highly proliferating but slowly growing paucicellular tumor. High Ki-67 expression does not seem to be an adverse prognostic factor in pediatric and adolescent patients with Hodgkin's lymphoma treated by effective risk-adapted chemo-radiotherapy regimens.