Favorable Prognosis In Breast Cancer Research Articles

Human Solid Tumors Contain High Endothelial Venules: Association with T- and B-Lymphocyte Infiltration and Favorable Prognosis in Breast Cancer

The mechanisms governing infiltration of lymphocytes into tumors remain poorly characterized, in spite of the critical impact of these cells on patient prognosis and therapeutic responses. High endothelial venules (HEV) are blood vessels found in lymphoid tissues, specialized in lymphocyte recruitment, but their implications in human cancer are unknown. In this article, we report the presence of MECA 79(+) blood vessels displaying all the phenotypic characteristics of HEVs in most of the 319 human primary solid tumors, including melanomas, breast, ovarian, colon, and lung carcinomas, analyzed. Tumor HEVs were specifically located within lymphocyte-rich areas, and their density within the tumor stroma was a strong predictor of infiltration by CD3(+) and CD8(+) T cells as well as B cells. Large-scale flow cytometric and quantitative reverse transcriptase-PCR analyses in freshly operated breast tumors revealed that high densities of tumor HEVs correlated with increased naive, central memory and activated effector memory T-cell infiltration and upregulation of genes related to T-helper 1 adaptive immunity and T-cell cytotoxicity. Finally, in a retrospective cohort of 146 invasive breast cancer patients, we found that high densities of tumor HEVs independently conferred a lower risk of relapse and significantly correlated with longer metastasis-free, disease-free, and overall survival rates. Together, our findings suggest that tumor HEVs function as major gateways for lymphocyte infiltration into human tumors, and may represent attractive targets for cancer diagnosis and therapy.

CXCR4 alters ID family and ITF2B/E2-2/TCF-4 dynamics to confer invasive phenotype to breast cancer cells.

Abstract Abstract #2045 Introduction: Breast cancer is characterized by a distinct metastatic pattern involving lymph nodes, bone marrow, lung, liver and brain. The chemokine CXCL12 and its receptor CXCR4 are suggested to play a role in invasion and organ-specific metastasis. Signaling pathways downstream of CXCL12:CXCR4 involved in invasion and/or metastasis are not known.
 Methods: MDA-MB-231 breast cancer cells were sorted for CXCR4-positive and CXCR4-negative subpopulation by FACS and subjected to microarray gene expression analysis. Differentially expressed genes were further verified by RT-PCR or qRT-PCR. siRNA or shRNA expression studies were used to evaluate the role of genes overexpressed in CXCR4-positive cells in invasion through Matrigel. Orthotopic xenograft model is being used to evaluate primary tumor growth and metastasis pattern of CXCR4-positive and CXCR4-negative subpopulation with various manipulations.
 Results: ITF2B, a class I bHLH transcription factor, is expressed at ∼4-fold higher level in CXCR4-positive cells. ITF2B mRNA level was lower in CXCR4 siRNA treated or CXCR4shRNA expressing CXCR4-positive cells suggesting that ITF2B is the direct target of CXCL12:CXCR4. ITF2B siRNA reduced invasion of CXCR4-positive cells whereas ITF2B overexpression increased invasive capacity of CXCR4-negative cells. ITF2B is a Wnt β-catenin regulated gene with transforming ability and controls the function of Inhibitor of DNA binding family of transcription factors (ID1 to ID4). ID1 is essential for lung metastasis whereas ID2 induces differentiation of breast cancer cells and higher levels of ID2 correlates with favorable prognosis in breast cancer. ID2 expression was higher in CXCR4-negative cells and its overexpression in CXCR4-positive cells reduced invasion. Both CXCR4 and ITF2B are involved in reducing ID2 expression as siRNA against both of these genes increased ID2 mRNA in CXCR4-positive cells. Interestingly, CXCR4-negative cells appear to have adapted to alternative mode of invasion/metastasis as these cells express higher levels of metastasis-associated genes such as TWIST and MT1-MMP. Xenograft studies with cells overexpressing ITF2B and ID2 are currently underway.
 Conclusions: CXCL12:CXCR4 activated signaling pathways target ID family/bHLH transcription factor network to promote invasion of breast cancer cells. Inducers of ID2 may help to overcome CXCR4- mediated invasion and/or metastasis of cancer cells. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2045.

Expression and Signaling Activity of Wnt-5a/Discoidin Domain Receptor-1 and Syk Plays Distinct but Decisive Roles in Breast Cancer Patient Survival

The loss of Wnt-5a, a G-protein-coupled receptor ligand, or Syk, an intracellular kinase, has in separate studies been associated with poor prognosis of breast cancer patients. Both proteins are involved in cell adhesion, a key event in epithelial cancer metastasis. Here, we have investigated whether Syk is part of the Wnt-5a/discoidin domain receptor-1 (DDR1) signaling pathway and if a signaling interaction of these proteins is important for breast cancer-specific survival. The signaling interactions between Wnt-5a/DDR1 and Syk were addressed in mammary cell lines. Their mRNA and protein levels and the respective clinical correlates were investigated in 94 cases of primary breast cancer. The expression of Wnt-5a and Syk correlated in four of five tumor cell lines. However, despite a constitutive association between Syk and the Wnt-5a-dependent adhesion receptor DDR1, we found no evidence of a Wnt-5a/DDR1-mediated activation of Syk. Instead, beta(1) integrins initiate the adhesion-induced activation of Syk. In tumors from breast cancer patients, the protein expression of Wnt-5a and Syk were differently regulated at the translational and transcriptional level, respectively. Analysis of breast cancer-specific survival revealed that the presence of Wnt-5a and Syk in primary tumors has good predictive value for a favorable outcome. Intriguingly, a simultaneous loss of both proteins did not reduce survival more than loss of either. Despite the difference in regulation of Wnt-5a and Syk protein expression and their lack of signaling interaction, our clinical data indicate that a favorable prognosis in breast cancer requires the expression and signaling activity of both.

Human kallikrein 13 protein in ovarian cancer cytosols: a new favorable prognostic marker.

Human kallikrein 13 (hK13; encoded by the KLK13 gene) is a secreted serine protease expressed in endocrine tissues, including the prostate, testis, breast, and ovary. We have previously reported steroid hormone regulation of the KLK13 gene and its clinical value as a marker of favorable prognosis in breast cancer at the mRNA level. We hypothesized that hK13 may represent a potential biomarker for ovarian carcinomas. Using a newly developed enzyme-linked immunosorbent assay (ELISA), hK13 levels were quantified in 131 ovarian tumor extracts and correlated with various clinicopathological variables and outcome (progression-free survival [PFS], overall survival [OS]), over a median follow-up period of 42 months. hK13 concentration in ovarian tumor cytosols ranged from 0 to 18.4 ng/mg of total protein. An optimal cutoff value of 0.13 ng/mg (67(th) percentile) was selected, based on the ability of hK13 values to predict the PFS of the study population, to categorize tumors as hK13-positive or negative. Women with hK13-positive tumors most often had early stage (stage I/II) disease, no residual tumor after surgery and optimal debulking success (P <.05). Univariate and multivariate Cox regression analyses revealed that patients with hK13-positive tumors had a significantly longer PFS and OS than hK13-negative patients (P <.05). Kaplan-Meier survival curves further confirmed a reduced risk of relapse and death in women with hK13-positive tumors (P =.007 and P =.002, respectively). These results indicate that hK13 is an independent marker of favorable prognosis in ovarian cancer.

Motility-related protein-1/CD9 expression in head and neck squamous cell carcinoma.

Motility-related protein (MRP)-1/CD9 is implicated in cell adhesion and motility and was shown to be clearly involved in tumor prognosis and angiogenesis. Elevated MRP-1/CD9 expression on tumor cells has been linked to a favorable prognosis in breast cancer, colon cancer, lung cancer, and HNSCC. Because MRP-1/CD9 is associated with angiogenesis, it might play a role in tumor angiogenesis as well. We analyzed MRP-1/CD9 expression in HNSCC specimens and cell lines by real-time RT-PCR and in HNSCC biopsy specimens and stromal vessels by immunohistochemistry. Kruskal Wallis and Chi2 test, univariate and multivariate Cox regression, and Kaplan-Meier methods were used for statistical analysis. Real-time and PCR RT showed elevated expression of MRP-1/CD9 in one (SCC25) of four HNSCC cell lines and two of six HNSCC patients, whereas two cell lines (SCC9 and JPPA) and one HNSCC patient had lower MRP-1/CD9 levels compared with other specimens. Immunohistochemistry demonstrated strong MRP-1/CD9 IR expression on tumor cells in 13 patients (39%), whereas 21 patients (61%) had less to medium MRP-1/CD9 IR expression. Increased MRP-1/CD9 expression on tumor cells was correlated with prolonged patient survival (p =.02) and a longer disease-free interval (p =.004), a diminished recurrence rate (p =.02), and lower stages of neck lymph nodes (p =.04). MRP-1/CD9 IR was also found in a subpopulation of vessels that seem to be less in tumor specimens than in normal mucosa (p <.0001). MRP-1/CD9+ vessels are podoplanin+ and are therefore regarded as lymphatic vessels. Our results revealed that elevated MRP-1/CD9 expression on HNSCC is linked to a favorable clinical outcome and confirmed reports of MRP-1/CD9 expression in other carcinomas. MRP-1/CD9+ vessels were found to be lymphatic in nature. The number and staining intensity of these vessels is decreased in tumor tissue, which suggests a stabilizing role for this protein in lymphangiogenesis.

The prognostic value of the human kallikrein gene 9 (KLK9) in breast cancer.

Many members of the human kallikrein gene family were found to be differentially expressed in various malignancies and some of them are useful diagnostic/prognostic markers. KLK9 is a newly discovered human kallikrein gene that is expressed in several tissues including thymus, spinal cord, testis, prostate, breast, and ovary. Like other kallikreins, the KLK9 gene was found to be regulated by steroid hormones, mainly estrogens and progestins, in cancer cell lines. We studied the expression of KLK9 by quantitative RT-PCR in 169 breast cancer patients of different stages, grades and histological types. We also compared the relation between KLK9 expression and other clinicopathological variables and patient survival. KLK9 expression is significantly higher in patients with early stage cancers (p = 0.039) and in patients with small tumor size (< 2 cm) (p = 0.028). Kaplan-Meier survival curves demonstrated that KLK9-positive patients have longer disease-free and overall survival (p = 0.015 and 0.036, respectively). Univariate and multivariate analysis also indicates that KLK9 expression is associated with increased disease-free and overall survival. When the Cox proportional hazard regression analysis was applied to subgroups of patients, KLK9 expression was found to be a significant predictor of disease-free survival in the estrogen receptor (ER) and progesterone receptor (PR) negative subgroups of patients (Hazard Ratio 'HR' = 0.28, and 0.38, respectively, and p = 0.011 and 0.028, respectively). After adjusting for other known prognostic variables, KLK9 retained its independent prognostic value in these subgroups of patients. Similar results were obtained for overall survival. KLK9 is a new potential independent marker of favorable prognosis in breast cancer.

Expression and clinical significance of pepsinogen C in epithelial ovarian carcinomas

Background: Pepsinogen C (pep C) is a gastric aspartic protease of which is associated with a favorable prognosis in breast cancer. Recently, it has been demonstrated in other tumors of extradigestive origin. Study design: We have analyzed pep C expression in 72 epithelial ovarian carcinomas by immunohistochemistry. Results: Nineteen (26%) tumors stained positively for pep C. Overall this expression was not associated with the clinicopathologic characteristics or with outcome. However, in patients with serum levels of CA 125 less than 35 U/ml, pep C expression identified a group with a more favorable prognosis. Conclusion: Pepsinogen C is expressed in a quarter of ovarian carcinomas and might identify a subset of patients with different prognosis.

Apolipoprotein D expression in endometrial carcinomas.

Apolipoprotein D is a protein component of the human plasma lipid transport system but is also associated with a more favorable prognosis in breast cancer and ovarian cancer women. This study was undertaken to examine the tumoral expression of apolipoprotein D in endometrial cancer and to analyze the possible correlation with tumor and patients characteristics as well as androgen receptors and its prognostic significance. Immunohistochemical evaluation was used to examine apolipoprotein D expression in paraffin blocks from 58 endometrial carcinomas. A total of twenty (34%) tumors stained positively. Staining was localized in tumor cells. No significant correlation was found between apo D expression and patients or tumor characteristics and androgen receptor status. In addition, apolipoprotein D expression was not associated with patient prognosis. Apolipoprotein D is expressed by a significant percentage of endometrial carcinomas without apparent association with other clinicopathologic parameters or with outcome of patients.

Identification of subgroups with favorable prognosis in breast cancer.

The prognostic significance of 16 factors was investigated in a series of 311 women with unilateral, invasive breast carcinoma without distant metastases (M0) and treated with mastectomy. The series consisted of 93% of such cases diagnosed histologically in the city of Turku, Finland, from 1980 to 1984. Mitotic count, histological and nuclear grades, extent of tumor necrosis, axillary nodal status (pN), tumor size (pT), estrogen and progesterone receptor (PR) contents, and S-phase fraction were the most powerful single factors (p = 0.0001 or less). Axillary node negative cancers with no or only spotty tumor necrosis (92% of all pN0 cases) were associated with a 96% 5-year survival rate corrected for intercurrent causes. Among the axillary node positive cases the combination of high PR content (greater than 60 fmol/mg protein) and a low mitotic count could identify a subgroup with a 96% 5-year corrected survival rate (25% of all pN+ cases). It is concluded that both pN0 and pN+ breast cancer contain identifiable subgroups with greatly different prognosis.

Morphological evidence for immune response to breast cancer. An historical review.

An historical review of morphological evidence for immune response to breast cancer is presented. At present it seems as if the closest the morphologist may come to seeing immunity is to see hyperplasia of some elements such as the round cells in tumors or the node changes or the immune system. Evidence now seems overwhelming that histocytosis comes before the good prognosis of less metastatic tumor in the axilla. In fact Black has found greater histiocytic reaction with in situ carcinoma than with invasive carcinoma. That is if the host immunity is great enough invasion may not be permitted even though cancer cells survive in the semiisolated environment of the breast ducts. Excess node tissue showing prominent histocytosis is suggestive of systemic reaction and good prognosis. The work of Black and Leis supports the original suggestion that histocytosis is related to cell-mediated immunity rather than to circulating antibodies.