Peritumoral Lymphatic Vessel Invasion Research Articles

Peritumoral lymphatic vessel density and invasion detected with immunohistochemical marker D240 is strongly associated with distant metastasis in breast carcinoma

BackgroundDetection of vascular invasion by hematoxylin and eosin staining is the current pathological assessment practice to diagnose breast carcinoma. However, conventional hematoxylin and eosin staining failed to distinguish between blood vessel invasion and lymphatic vessel invasion. Both are important prognostic criteria however with different outcomes. The aim of this study is to distinguish between blood vessel invasion and lymphatic vessel invasion using conventional assessment and immunohistochemical markers. The prognostic significance of both circulatory invasions in invasive breast carcinoma was also investigated.MethodsConsecutive sections of breast carcinoma samples from 58 patients were stained with CD34 and D240 to stain blood and lymphatic vessels respectively. Hematoxylin and eosin staining was carried out on another consecutive section as conventional staining.ResultsAlthough blood vessel density is higher in the sections (median = 10.3 vessels) compared to lymphatic vessel density (median = 0.13), vessel invasion is predominantly lymphatic invasion (69.8 and 55.2% respectively). Interestingly, peritumoral lymphatic vessel density and peritumoral lymphatic invasion was significantly associated with distant metastasis (p = 0.049 and p = 0.05 respectively). The rate of false positive and false negative interpretation by hematoxylin and eosin was 46.7 and 53.3% respectively.ConclusionsLymphatic vessel invasion is a strong prognostic markers of breast carcinoma invasion and the use of immunohistochemical markers increase the rate and accuracy of detection.

THORACIC SURGERY Intratumoral lymphatic vessel density, and intratumoral and peritumoral lymphatic vessel invasion as predictive factors of lymph node metastasis and prognostic factors in esophageal cancer

In esophageal cancer, metastases in regional lymph nodes occur early and are a poor prognostic factor. We studied intratumoral and peritumoral lymphatic vessel density, and intratumoral and peritumoral lymphatic vessel invasion using D2-40 immunohistochemical staining in 74 resected esophageal cancer specimens and correlated results with patients’ clinicopathologic features and outcomes. Results were analyzed by univariate and multivariate logistic regression as well as univariate and multivariate survival analysis. The peritumoral lymphatic vessel density was significantly higher than the intratumoral lymphatic vessel density (p < 0.001). The peritumoral lymphatic vessel density and intratumoral lymphatic vessel density positively correlated with lymph node metastasis, tumor stage, depth of tumor invasion, and residual tumor. Positive intratumoral lymphatic vessel invasion and peritumoral lymphatic vessel invasion were significantly correlated with lymph node metastasis, tumor size, tumor depth, stage, and residual tumor. Multivariate logistic regression analysis identified intratumoral lymphatic vessel density (p = 0.028) and peritumoral lymphatic vessel invasion (p = 0.036) as risk factors of lymph node metastasis. Cox regression analysis showed that intratumoral lymphatic vessel invasion and peritumoral lymphatic vessel invasion are independent prognostic factors of disease-free survival (p = 0.018, p = 0.021), cancer-specific survival (p = 0.016, p = 0.025), and overall survival (p = 0.017, p = 0.02). Intratumoral lymphatics and peritumoral lymphatic vessel invasion are predictive factors of lymph node metastasis. Intratumoral and peritumoral lymphatic vessel invasion are independent prognostic factors in locally advanced esophageal cancer.

Significance of intratumoral lymphatic vessel density and peritumoral lymphatic vessel invasion identified by D2-40 in patients with esophageal cancer

Significance of intratumoral lymphatic vessel density and peritumoral lymphatic vessel invasion identified by D2-40 in patients with esophageal cancer

Clinical significance of peritumoral lymphatic vessel density and lymphatic vessel invasion detected by D2-40 immunostaining in FIGO Ib1-IIa squamous cell cervical cancer

The clinical significance of lymphangiogenesis in cervical cancer remains controversial. Our aim was to investigate the correlation between lymphangiogenesis, lymphatic vessel invasion (LVI) and tumor metastasis, invasion and prognosis in squamous cell cervical cancer. Paraffin sections of 90 patients with FIGO (Fédération Internationale de Gynécologie et d'Obstétrique) Ib1-IIa squamous cell cervical cancer were stained for immunohistochemistry with a D2-40 monoclonal antibody against the carcinoembryonic antigen M2A. The lymphatic vessel density (LVD) and LVI were measured, and their relationship with the clinicopathological data was analyzed. D2-40-positive lymphatic vessels were found in 75 of the 90 patients (83.3 %). All D2-40-positive vessels were located in peritumoral areas. The mean±SD of the peritumoral LVD was 10.08±4.16. The positive rate of LVI was 32.0 % (24/75). The recurrence rate of patients with LVD >10 (62.1 %, 18/29) was significantly higher than that of patients with LVD ≤10 (34.8 %, 16/46, P = 0.021). The 5-year recurrence-free survival rate of patients with LVD >10 (41.0 %) was significantly lower than that of patients with LVD ≤10 (67.0 %, P = 0.045). Univariate analysis showed that the peritumoral LVD (≤10 vs >10) was correlated with LVI (absent vs present, P = 0.016). The peritumoral LVD and LVI showed no correlation with age, FIGO stage, tumor size, tumor grade, depth of invasion, or pelvic lymph node metastasis (all: P > 0.05). Peritumoral lymphangiogenesis was correlated with the recurrence and recurrence-free survival in patients with squamous cell cervical cancer. Examination of peritumoral LVD in these patients might therefore help to estimate the risk of recurrence.

Intratumoral as well as peritumoral lymphatic vessel invasion correlates with lymph node metastasis and unfavourable outcome in colorectal cancer

The aim of this study was to assess the intratumoral and peritumoral distribution of lymphatic vessel density (LVD) and lymphatic vessel invasion (LVI) in colorectal cancer and their relationships with patients' clinicopathological characteristics and survival. Paraffin sections of 81 primary colorectal cancers were examined by immunohistochemical staining using monoclonal antibody D2-40. Peritumoral LVD was significantly higher than intratumoral LVD (P = 0.000). Both intratumoral LVD and peritumoral LVD were correlated with the presence of LVI (P = 0.006 and P = 0.003, respectively). LVI, intratumoral LVI and peritumoral LVI were identified, respectively in 38, 28 and 32% of the samples investigated. Both intratumoral LVI and peritumoral LVI were correlated with lymph node metastasis (P = 0.030 and P = 0.014, respectively). Lymph node metastasis, the presence of intratumoral LVI and peritumoral LVI were adversely associated with the 5-year overall survival in a univariate analysis (P = 0.001, P = 0.011 and P = 0.017, respectively). Multivariate analysis using Cox proportional hazard model showed that neither intratumoral LVI nor peritumoral LVI was an independent prognostic factor of overall survival. The results of this study demonstrated that intratumoral as well as peritumoral LVI was associated with lymph node metastasis and adverse outcome in colorectal cancer.

Vascular endothelial growth factor-A and -C: expression and correlations with lymphatic metastasis and prognosis in colorectal cancer

Previous studies obtained contradicting results regarding the correlation between expression of VEGF-A, VEGF-C and colorectal cancer patients' clinicopathological features and prognosis. Moreover, the association between the growth factors' expression and lymphatic vessel invasion (LVI) with intratumoral and peritumoral difference has not been reported. In this study, 81 primary colorectal cancer samples were immunohistochemically stained for VEGF-A, VEGF-C and podoplanin. The expression of VEGF-A and VEGF-C in marginal portion was significantly higher than those in central portion (P = 0.000 for both). The expression of VEGF-A in marginal portion was correlated with lymph node metastasis (P = 0.031). The expression of VEGF-C in marginal portion was correlated with TNM stage (P = 0.045), peritumoral LVI (P = 0.048) and lymph node metastasis (P = 0.019). The group with high VEGF-A expression in marginal portion showed worse survival than the low expression group (P = 0.039). Patients with high expression of VEGF-C in the marginal portion were not significantly different from those with low VEGF-C expression (P = 0.121). Patients with high expression of both VEGF-A and VEGF-C in the marginal portion showed the worst survival (P = 0.015). In conclusion, increased expression of VEGF-A and VEGF-C in marginal portion of colorectal cancer was correlated with lymph node metastasis. VEGF-C facilitates colorectal cancer cells invade into peritumoral lymphatic vessels, and different mechanisms may exist in the invasion of tumor cells into peritumoral and intratumoral lymphatic vessels. Assessment the expression of both VEGF-A and VEGF-C in the marginal portion of tumor may help to identify patients with colorectal cancer with unfavourable overall survival.

Prognostic Impact of Marginal Adipose Tissue Invasion in Ductal Carcinoma of the Breast

This study aimed to investigate whether adipose tissue invasion (ATI) of cancer cells at the tumor margin influenced lymph node status and prognosis in patients with invasive ductal carcinoma of the breast. Data for 245 patients with breast cancer with marginal ATI were clinicopathologically compared with data for 65 patients without ATI. We also examined the value of the combination of ATI and peritumoral lymphatic vessel invasion (LVI). The frequency of axillary lymph node metastases was 40.7% in patients with ATI (99/243) and 11.3% in patients without ATI (7/62; P<.0001), and ATI was an independent factor influencing nodal metastasis. Patients with ATI had a poorer prognosis than patients without ATI (10-year disease-free survival, 76% and 94%, respectively; P=.0323). In addition, patients without ATI or LVI had neither lymph node metastasis (n=52) nor recurrent disease (n=53). ATI is one of the biologic indicators of tumor aggressiveness.

The Role of Lymphovascular Invasion in Predicting the Prognosis of Clinically Localized Upper Tract Urothelial Carcinoma (pT1-3cN0M0)

We assessed the impact of lymphovascular invasion on the prognosis of patients treated for clinically localized (pT1-3N0M0) upper tract urothelial carcinoma. From January 1998 to December 2004, 106 patients treated surgically for clinically localized upper tract urothelial carcinoma (pT1-3cN0M0) were recruited for analysis retrospectively. We assessed the impact of multiple prognostic factors including age, sex, smoking, tumor multiplicity, pathological stage, grading, squamous differentiation, subsequent bladder tumor recurrence, tumor site and lymphovascular invasion on the recurrence-free survival rate. We clarified the role of lymphovascular invasion in predicting cancer specific survival in these patients. Mean patient age was 67.2 +/- 11.1 years and median followup was 47.5 months (range 40 to 115). Lymphovascular invasion was present in 32 of 106 (30.2%) patients. The 5-year recurrence-free survival rate in patients with and without lymphovascular invasion was 65.3% and 91.9%, respectively (p <0.001). On multivariate analysis lymphovascular invasion status, pT3 and ureteral involvement were significant predictors of patient recurrence-free survival. The positivity of lymphovascular invasion was also significantly associated with the risk of higher stage disease (OR 7.49). Patients with lymphovascular invasion had a higher risk of greater pathological stage disease. Lymphovascular invasion and high stage disease with ureteral involvement led to a greater risk of disease recurrence and, in turn, recurrence caused a higher mortality rate. This finding could help identify patients at greater risk for disease recurrence who would benefit from close followup and early adjuvant therapy.

Lateral peritumoral lymphatic vessel invasion can predict lymph node metastasis in esophageal squamous cell carcinoma

Lymph node metastasis is an important prognostic factor in many types of cancer. We investigated the clinical significance of lymphangiogenesis and lymphatic vessel invasion in esophageal squamous cell carcinoma. We evaluated lymphatic vessel density and lymphatic vessel invasion in the intratumoral, peritumoral and normal compartments using D2-40 immunostaining. In addition, the peritumoral compartment was divided into the lateral peritumoral compartment and the non-lateral peritumoral compartment. The lymphatic vessel density was higher in the peritumoral and intratumoral compartments than in the normal compartment. However, the lymphatic vessel density did not correlate with any pathological parameters including lymph node metastasis. Intratumoral and peritumoral lymph vessels were small and collapsed while normal lymphatic vessels and lymphatic vessels with lymphatic vessel invasion were dilated and large. The presence of lymphatic vessel invasion, in the lateral peritumoral compartment but nowhere else, significantly correlated with lymph node metastasis. These results suggest that lymphangiogenesis might occur with esophageal cancer, but it does not play a direct role in lymphatic vessel invasion and lymph node metastasis. Peritumoral lymphatic vessel invasion, especially in the lateral peritumoral compartment, should imply a high probability of regional lymph node metastasis.

Peritumoral lymphatic invasion is associated with regional lymph node metastases in prostate adenocarcinoma

Lymphangiogenesis, detected by antibodies specific for lymphatic endothelial cells, has been associated with regional lymph node metastases and poor prognosis in carcinomas of head and neck, breast and uterine cervix, but remains largely uninvestigated in prostate adenocarcinoma. We evaluated the lymphatic vessel density and lymphatic vessel invasion by prostate cancer cells in the intratumoral, peritumoral and normal prostate tissue compartments in cancer-bearing prostate glands and correlated them with lymph node metastases, Gleason score and other pathological parameters. Lymphatic vessels were detected by immunohistochemical stain using an antibody specific for the lymphatic endothelial cells (clone D2-40) on 33 radical prostatectomies. In all, 26 patients had lymph node dissection, and 14 of them had lymph node metastasis. The lymphatic vessel density and lymphatic vessel invasion were then recorded for each of the three compartments microscopically. Lymphatic vessel density in the intratumoral, peritumoral and normal prostate compartments was 0.91±0.80, 1.54±0.68 and 1.58±0.96/mm2, respectively. The intratumoral lymphatic vessel density was significantly lower than that of the peritumoral and normal prostate compartments, and the latter two were not significantly different. The lymphatic vessel density of the three compartments was not significantly different between cases with and without lymph node metastasis. The peritumoral lymphatic vessel density correlated inversely with the Gleason score. Lymphatic vessel invasion was present in significantly higher percentage of cases with lymph node metastasis (9/14, 62.3%), as compared to those without lymph node metastasis (1/12, 8.3%, P<0.01). The peritumoral lymphatic vessel invasion had a better correlation with the presence of lymph node metastases than intratumoral lymphatic vessel invasion. There is no evidence of lymphangiogenesis in prostate adenocarcinoma. Peritumoral lymphatic vessel invasion correlates with regional lymph node metastases, suggesting that the peritumoral lymphatic vessels are functionally important and identification of lymphatic vessel invasion in this compartment implies a high probability of regional lymph node metastases.

Dependencia temporal del valor pronóstico de los receptores esteroides en el cáncer de mama

The influence of prognostic factors in breast cancer may change during follow-up. Our aim was therefore to look for time-related changes in hazard ratios for steroid receptor status and the risk of relapse. We studied 455 patients with operable primary breast cancer during a mean follow-up period of 72 months (range, 42-130 months). Clinical and histological characteristics were assessed and estrogen receptor (ER) and progesterone receptor (PgR) status were determined and compared to disease-free survival (DFS) and overall survival (OS). Analysis of DFS included fitting a multivariate Cox proportional hazards model, testing for non-proportionality, and examining diagnostic plots. A total of 66.8% of the tumors were ER positive and 54.7% PgR positive. Receptor status is highly associated with menopausal status, histological grade, peritumoral lymphatic or blood vessel invasion and mitotic index. No correlation was found between steroid receptor status and tumor size, node status or tumor stage. In univariate analysis, ER/PR vs ER+/PR+ display a hazard ratio of 2.15 (IC del 95%, 1.59-2.99) for DFS (p = 0.001) and 1.95 (IC del 95%, 1.38-2.59) for OS (p = 0.0043). In multivariate analysis, steroid receptor status, node status and mitotic index were independent prognostic factors for DFS and OS. By using Cox regression time-dependent covariates model, we show that the steroid receptor status hazard ratio is 5.6 at diagnosis, decreases after 4 years, and loses its significance after 10 years of follow-up. In patients with breast cancer the status steroid receptor have a relatively limited prognostic value and is lost within the first years.

Preoperative values of CA 15-3 and CEA as prognostic factors in breast cancer: a multivariate analysis.

The role of circulating tumor markers in providing prognostic information has not been widely studied. In the current study, serum levels of the carbohydrate antigen 15-3 (CA 15-3) and carcinoembryonic antigen (CEA) were determined preoperatively in 364 breast cancer patients with no clinical signs of metastasis. The prognostic relevance of these markers for recurrence (175/364) and death of disease (104/175) was determined by Cox multivariate analysis, including the comparison with classical prognostic factors. High levels of both tumor markers were associated with aneuploid tumors with high S-phase fraction and high ornithine decarboxylase activity. CA 15-3 was highly associated with the number of positive lymph nodes and peritumoral lymphatic or blood vessel invasion. No significant associations were found between CEA or CA 15-3 levels and histologic grade, necrosis and steroid receptor status. In univariate analysis, preoperative values, using optimum cutoff values of CA 15-3 (40 U/ml) and CEA (6 ng/ml), were statistically significant for relapse-free survival and overall survival. In multivariate analysis, only node status, DNA ploidy and ornithine decarboxylase activity were independent predictors for relapse-free survival; the estrogen receptor status was a predictor of overall survival. In node-negative patients, ornithine decarboxylase activity was the only factor selected for relapse-free survival. In node-positive patients, the number of lymph nodes and DNA ploidy were the only variables selected for relapse-free survival or overall survival. Estrogen receptor and ornithine decarboxylase activity were excluded for relapse-free survival, but were significant prognostic factors for overall survival.

Bone metastasis after a resection of stage I and II primary lung cancer

In the present study, we reviewed the patients who developed bone metastases after a surgical resection of primary lung cancer and evaluated their clinicopathological features. From 1992 to 1995, 177 patients with stage I and II primary lung cancer underwent a surgical resection at the Kitakyushu Municipal Medical Center. Bone metastases were detected in 14 patients (7.9%) by follow-up examinations including bone scintigraphy (scan). Bone metastasis was one of the most frequent extra-thoracic recurrent forms. Patients with adenocarcinoma tended to develop bone metastases more frequently than those with squamous cell carcinoma. In the preoperative bone scans, an abnormal uptake was observed in 76 patients (42.9%), and 10 (13.1%) of them were found to develop bone metastases in the follow-up studies. A microscopic examination of the primary tumor demonstrated close correlation between intratumoral and peritumoral lymphatic vessel invasion and postoperative development of bone metastases. A bone scan is a very useful and indispensable procedure for diagnosing bone metastases. However, this scan may also show false positive finding in a number of benign conditions. Therefore, a surgical resection should be considered as the first-line treatment for patients with positive findings in the bone scan when the diagnosis of bone metastasis can not be confirmed based on both their symptoms and other clinical examinations.

Fatty acid synthase (FAS) predictive strength in poorly differentiated early breast carcinomas.

Many normal and human cancer tissues express fatty acid synthase (FAS), the major enzyme required for endogenous fatty acid biosynthesis. Strong expression of FAS seems to be associated with a poor prognosis. This study examines the strength of FAS and other common markers of relapse in poorly differentiated breast carcinoma. Fifty-one patients with poorly differentiated ductal infiltrating breast carcinomas were followed up for more than 10 years. Immunohistochemical detection of FAS was associated with morphological features of the tumors, with immunohistochemical expression of c-erbB-2, cathepsin D, estrogen and progesterone receptor status and with DNA ploidy in order to detect a statistical correlation. The chi-square test revealed a correlation between FAS and peritumoral lymphatic vessel invasion (PLVI) (P = 0.001). Univariate analysis showed that FAS was correlated with disease-free survival (DFS) (P = 0.0001). Other prognosticators associated with DFS were PLVI (P = 0.002), estrogen (P = 0.008) and progesterone receptor status (P = 0.007). Bivariate analysis showed that FAS was a further prognostic discriminant of DFS within the ER, PgR and PLVI subsets. FAS is a reliable prognosticator of recurrence in poorly differentiated early breast carcinomas. Association of FAS with PLVI may be useful to plan a correct follow-up in patients with breast neoplasms.

Prognostic Singnificance of Peritumoral Lymphatic Vessel Invasion in Breast Cancer

Prognostic Singnificance of Peritumoral Lymphatic Vessel Invasion in Breast Cancer

Expression of fatty acid synthase (FAS) as a predictor of recurrence in stage I breast carcinoma patients.

Fatty acid synthase (FAS) is a molecule found in tumor cells from breast carcinomas of patients whose prognosis is very poor. Recently, this molecule has been identified as the key enzyme in fatty acid biosynthesis. This study was done to test the strength of FAS as a prognostic indicator for disease free survival (DFS) and overall survival (OS). Clinical records, histologic features, immunohistochemical expression of cathepsin D and c-erbB-2, and estrogen and progesterone receptor status of 110 Stage I breast carcinoma patients were all associated with FAS by a chi-square test. The patterns of DFS and OS were estimated over a ten-year follow-up period using the Kaplan-Meier method. Univariate and multivariate analysis were evaluated using a log logistic regression model. Multivariate regression analysis was based on the Cox proportional hazard model. To detect FAS, cathepsin D and c-erbB-2 expression as well as estrogen and progesterone receptor status, we used the unlabeled immunoperoxidase technique on formalin fixed, paraffin embedded tissue. FAS was significantly associated with a higher risk of recurrence because it predicted both DFS (P = 0.0001) and OS (P = 0.003) when evaluated as a continuous variable and DFS (P = 0.0001) when evaluated with other prognostic markers. Peritumoral lymphatic vessel invasion was the other most significant independent predictor for DFS (P = 0.001) and OS (P = 0.003). FAS is a reliable prognostic marker to predict DFS and OS in patients with early breast cancer.

34 P - Blood vessel invasion by tumor cells and tumor angiogenesis could predict recurrence in resected non shall cell lung cancer

Traditional parameters such aa tumor size, histology, hilar and mediastinal lymph nodes involvement have been identified as prognostic faeton of natural history of completely resected lung cancer. New factors including tumor proliferative activity have been evaluated in the research setting, but at the moment their prognostic significance in resected lung cancer is not identified. Therefore controversy continues as to whether tumor proliferative index provides more useful clinical information than traditional prognostic parameters. We investigated parameters like proliferative activity, fibrosis and necrosis, mitotic count, intratumoral and peritumoral blood or lymphatic vessel invasion by tumor cells and neoangiogenesis in 125 consecutive patients with non small cell lung cancer and who underwent operation between January 1990 and December 1993. Seventy-four were in stage I, 13 in stage II and 38 in stage III, 52 were adenocarcinoma, 55 squamous cell carcinoma, 11 adeno-squamous cell carcinoma and 7 indifferentiated large cell carcinoma. Fourteen patients in stage III had previous neoadjuvant chemotherapy with 3 cycles of mitomycin, cispiatin and vinbiastine (MPV) and after underwent surgery. Median follow-up of the entire group is 3.4 years. Fifty-three patients in stage I (71.6%) are still alive and free of disease; eight patients (61.5%) in stage II and 20 (53.6%) in stage III are alive respectively. Eleven patients (76.6%) who had previous chemotherapy and complete resection are alive and free of disease at a median follow-up of 22 months. By multivariate analysis the disease-free survival seems to be influenced by blood vessel invasion by tumor cells and neoangiogenesis expecially for patients in stage I and in stage III for neoadjuvant group. In this latter group exists a correlation between fibrosis grade, presence of inflammatory peritumoral reaction and survival. Preliminary data of our study show a correlation between either the density of microvesssls or the blood vessel invasion in histologic sections and the recurrence of disease. Furthermore our results suggest that the presence of concomitant fibrosis and inflammatory infiltration could be an important factor for better clinical out-come althought longer follow-up is needed to assess the impact of this multimodality evaluation.

67‐kDa laminin‐receptor expression adds prognostic information to intra‐tumoral microvessel density in node‐negative breast cancer

Experimental studies have shown that the 67-kDa laminin receptor (LRec) is an important molecule for the interaction of tumour cells with the extracellular matrix, and that it plays a role in the early steps of angiogenesis and in tumour invasion and metastasis. We performed a multi-parametric study in 171 node-negative breast cancers, followed for a median time of 6 years, to verify whether determination of the LRec provides prognostic information in addition to intra-tumoral microvessel density (IMD), a measure of tumour angiogenesis, and to other biological and conventional indicators. A positive association between LRec expression and high neovascularization was found, although it did not reach significance. LRec was not associated either with other biological markers (oestrogen receptor, progesterone receptor and p53 expression) or to the conventional prognostic features [menopausal status, tumour size, histological types, grading and peri-tumoral lymphatic vessel invasion (PLVI)]. In univariate analysis, IMD, p53, PgR, PLVI, grading and tumour size were significant prognostic indicators of relapse-free survival (RFS). LRec expression approached significance when considered as a dichotomous variable, after having selected the optimum cutoff of 10% to distinguish high-risk from low-risk patients. For overall survival (OS), tumour size and IMD (continuous variable) were significant prognostic factors, and p53 approached significance. In multivariate analysis for RFS, the joint variable LRec and vascularization was the strongest independent prognostic factor, followed by PgR, PLVI and p53. For OS, tumour size was the only independent prognostic indicator in this series.

Prognostic value of intratumoral microvessel density, a measure of tumor angiogenesis, in node-negative breast carcinoma--results of a multiparametric study.

In the present study we update previous results on the prognostic value of intratumoral microvessel density (IMD), determined immunocytochemically using the monoclonal antibody CD-31 and a standard streptavidin-immunoperoxidase technique, published in the J Clin Oncol 12:454-466, 1994. This study was undertaken in those 211 node-negative breast cancer (NNBC) cases of that series of which we had pathological material available to determine all the prognostic indicators. The median period of follow-up has been extended to 78 and 80 months for relapse-free survival (RFS) and overall survival (OS), respectively, and new biological indicators (i.e. Ki-67 labeling and 67 kDa laminin receptor expression) were included in the analysis. The main results obtained are: i) a confirmation that IMD is not associated with the other biological markers studied, i.e. expression of p53 protein, c-erbB-2 protein, 67 kDa laminin receptor, and cell kinetics; IMD was weakly associated only with histological grade (p = 0.053); ii) IMD remains a highly significant prognostic factor for RFS and OS (p < 0.0001 and p = 0.018, respectively) in univariate analysis; iii) in multivariate analysis on RFS, IMD (likelihood ratio test (LRT) = 30.16; p < 0.0001), 67 kDa laminin receptor (LRT = 9.80; p = 0.0017), the IMD/67 kDa laminin receptor interaction (LRT = 8.62; p = 0.0033), tumor size (LRT = 8.56; p = 0.0034), and p53 protein (LRT = 4.96; p = 0.025) are significant and independent prognostic indicators. For OS, only tumor size (LRT = 8.34; p = 0.0038), menopausal status (LRT = 5.16; p = 0.023), p53 protein (LRT = 4.37; p = 0.036), and IMD (LRT = 4.05; p = 0.044) retain a significant and independent prognostic value. The results of this study confirm the prognostic importance on RFS of the variables previously tested, but not of peritumoral lymphatic vessel invasion. A novel finding is that 67 kDa laminin receptor and the IMD/67 kDa laminin receptor interaction are also significant and independent variables. For OS, the results confirm that both IMD and tumor size are significant and independent variables. With prolonged follow-up the novel finding that emerges is the prognostic importance of menopausal status and p53 protein. This new information could be useful for a more accurate selection of high-risk NNBC patients who require careful follow-up and may benefit from adjuvant therapy.

Cell kinetics in human breast cancer: comparison between the prognostic value of the cytofluorimetric S-phase fraction and that of the antibodies to Ki-67 and PCNA antigens detected by immunocytochemistry.

The determination of cell proliferation is one of the more widely used tools for assessing prognosis. However, additional research in this field is warranted because today there are several methodological procedures available for monitoring cell kinetics and it has still not been established which is the most reliable marker of proliferation and which possesses the greatest prognostic value. We performed this study in a series of primary invasive breast cancers to compare the prognostic value of S-phase fraction (SPF) by flow cytometry, the most widely used method for detecting proliferation at present, with that of antibodies to Ki-67 and PC-10 to proliferating-cell nuclear antigen (PCNA) detected by immunocytochemical methods. A significant linear relationship was observed only between SPF and Ki-67. In univariate analysis SPF and Ki-67 values, nodal status, histological grading and peritumoral lymphatic-vessel invasion were significant predictors of relapse-free survival (RFS). As far as overall survival (OS) is concerned, only SPF, Ki-67 and nodal status were significantly associated with the risk of death. PCNA had no prognostic value for either RFS or OS. In multivariate analysis only SPF and nodal status retained a significant and independent prognostic value. Neither the cell-kinetics parameters assessed by immunocytochemistry (i.e. Ki-67 and PCNA) nor histological grading were independent prognosticators. In conclusion, our results provide evidence that the determination of SPF by flow cytometry was the strongest cell-kinetics marker used to assess prognosis in this series of breast cancers. However, different and novel markers of cell kinetics need to be compared in larger series in order to identify the best one.