MMP-1 expression has an independent prognostic value in breast cancer.

BACKGROUND: Breast cancer consists of a variety of tumours, which differ by their morphological features, molecular characteristics and outcome. Well-known prognostic factors, e.g. tumour grade and size, Ki-67, hormone receptor status, HER2 expression, lymph node status and patient age have been traditionally related to prognosis. Although the conventional prognostic markers are reliable in general, better markers to predict the outcome of an individual tumour are needed. Matrix metalloproteinase-1 (MMP-1) expression has been reported to inversely correlate with survival in advanced cancers. In breast cancer MMP-1 is often upregulated, especially in basal-type breast tumours. The purpose of this retrospective study was to analyse MMP-1 expression in breast cancer cells and in cancer associated stromal cells and to correlate the results with traditional prognostic factors including p53 and bcl-2, as well as to patient survival in breast cancer subtypes. METHODS: Immunohistochemical analysis of MMP-1, ER, PR, Ki-67, HER2, bcl-2, p53 and CK5/6 expression was performed on 125 breast cancers. Statistical analyses were carried out using Kruskal-Wallis and Mann-Whitney -tests. In pairwise comparison Bonferroni-adjustment was applied. Correlations were calculated using Spearman rank-order correlation coefficients. Kaplan-Meier survival analyses were carried out to compare breast cancer-specific survival curves. Factors significantly associated with disease-specific survival in univariate models were included in multivariate stepwise. RESULTS: Positive correlations were found between tumour grade and MMP-1 expression in tumour cells and in stromal cells. P53 positivity significantly correlated with MMP-1 expression in tumour cells, whereas HER2 expression correlated with MMP-1 both in tumour cells and stromal cells. MMP-1 expression in stromal cells showed a significant association with luminal A and luminal B, HER2 overexpressing and triple-negative breast cancer subtypes. CONCLUSIONS: The most important finding of this study was the independent prognostic value of MMP-1 as well as Ki-67 and bcl-2 expression in tumour cells. Our study showed also that both tumoural and stromal MMP-1 expression is associated with breast tumour progression and poor prognosis. A significant difference of MMP-1 expression by cancer associated stromal cells in luminal A, luminal B and triple-negative breast cancer classes was also demonstrated.

Expression of matrix metalloproteinase-1, -7, -9, -13, Ki-67, and HER-2 in epithelial-myoepithelial salivary gland cancer.

BACKGROUND: The expression of matrix metalloproteinases (MMPs) in epithelial-myoepithelial salivary gland carcinoma has not been studied previously. METHODS: Immunohistochemistry for MMP-1, -7, -9, -13, Ki-67, and HER-2, as well as HER-2 gene amplification by silver enhanced in situ hybridization was performed in a series of 12 paraffin-embedded histopathologic samples of patients from Canada and Finland. RESULTS: A positive MMP-13 (p = .0022), higher MMP-13 (p = .0274), and higher MMP-9 (p = .0274) index (multiplication of staining intensity by percentage of the positive cells) predicted better overall survival. In disease-specific analysis, higher MMP-9 index (p = .0327) predicted better survival. A higher volume corrected index (VCI) of Ki-67 (p = .0339) predicted worse disease-specific survival. In 1 patient, HER-2 oncogene amplification was observed. CONCLUSION: MMPs and Ki-67 may have prognostic impact in epithelial-myoepithelial carcinoma.

Matrix metalloproteinase (MMP)-7 in salivary gland cancer.

INTRODUCTION: High levels of certain matrix metalloproteinases (MMPs) have been detected in various human cancers. The purpose of this study was to analyze the expression of MMP-7 in salivary gland cancer (SGC) by immunohistochemistry and to associate the results with the clinical data and the 10-year survival of the SGC patients. MATERIAL AND METHODS: Immunohistochemistry for MMP-7 was performed in a series of 107 paraffin-embedded sections of SGC. The samples represent the entire SGC population in Finland from 1991-1996. Mortality follow-up ended December 31, 2006. RESULTS: The study population of 107 patients consisted of 47 male and 60 female subjects, ranging in age at the time of diagnosis between 23 and 90 years. The minimum follow-up time was 10.6 years and the maximum 15.9 years. By age-adjusted analysis lower staining intensity was associated with worse overall survival of patients with acinic cell carcinoma (p = 0.047, HR 6.5, 95% Cl 1.0-41.7) and in mucoepidermoid carcinoma (p = 0.010, HR 9.3, 95% CI 1.7-50.0). Low staining intensity was also associated with worse disease-specific survival of patients with acinic cell carcinoma (0-1 vs. 2-3; p = 0.047, HR 13.7, 1.0-200.0). VCI Ki-67 was an important prognostic factor for survival of the entire data set (p < 0.0001, HR 4.7, 95% Cl 2.3-9.8). CONCLUSIONS: MMP-7 is associated with the prognosis of patients with acinic cell and mucoepidermoid carcinoma.

Gene expression profiling of endometrial adenocarcinomas reveals increased apolipoprotein E expression in poorly differentiated tumors.

INTRODUCTION: Tumor grade is one of the most important prognostic factors in endometrioid endometrial adenocarcinoma. Amplification of oncogenes, such as Her2/neu, or loss of function of tumor suppressor genes, such as p53, are known to be associated with poor prognosis, but additional factors influencing clinical behavior are likely to exist. To examine the biological differences between low-grade and high-grade endometrioid endometrial adenocarcinomas, we compared gene expression in these 2 types of tumors. METHODS: Six well-differentiated adenocarcinomas and 7 poorly differentiated adenocarcinomas were studied with 2 different microarray platforms, Affymetrix and Illumina. The expression of the most differentially expressed gene on both platforms was further studied in 34 endometrial adenocarcinoma samples (10 well differentiated, 9 moderately differentiated, and 15 poorly differentiated) using real-time reverse transcription-polymerase chain reaction. RESULTS: The most differentially expressed gene on both platforms was Apolipoprotein E (APOE). In the poorly differentiated adenocarcinomas, APOE was overexpressed 13.1-fold (P = 0.001) and 9.7-fold (P = 0.007) when compared with well- and moderately differentiated tumors, respectively. There was no difference in APOE expression between well- and moderately differentiated adenocarcinomas. CONCLUSIONS: Increased expression of APOE might represent a late event in the progression of well-differentiated endometrioid endometrial adenocarcinoma to a poorly differentiated endometrioid endometrial adenocarcinoma. Although increased APOE expression has been previously reported in other malignancies, this is the first study to suggest that APOE might also have a role in endometrioid endometrial cancer.

Analysis of cyclins A, B1, D1 and E in breast cancer in relation to tumour grade and other prognostic factors.

BACKGROUND: The cell cycle is promoted by activation of cyclin dependent kinases (Cdks), which are regulated positively by cyclins and negatively by Cdk inhibitors. Proliferation of carcinoma is associated with altered regulation of the cell cycle. Little is known on the combined alterations of cyclins A, B1, D1 and E in breast cancer in relation to the tumour grade and other prognostic factors. FINDINGS: Immunohistochemical analysis of cyclins A, B1, D1 and E, estrogen receptor, progesterone receptor, Ki-67, Her-2/neu and CK5/6 was performed on 53 breast carcinomas. mRNA levels of the cyclins were analysed of 12 samples by RT-PCR. The expression of cyclins A, B1 and E correlated with each other, while cyclin D1 correlated with none of these cyclins. Cyclins A, B1 and E showed association with tumour grade, Her-2/neu and Ki-67. Cyclin E had a negative correlation with hormone receptors and a positive correlation with triple negative carcinomas. Cyclin D1 had a positive correlation with ER, PR and non-basal breast carcinomas. CONCLUSION: Cyclin A, B1 and E overexpression correlates to grade, Ki-67 and Her2/neu expression. Overexpression of cyclin D1 has a positive correlation with receptor status and non-basal carcinomas suggesting that cyclin D1 expression might be a marker of good prognosis. Combined analysis of cyclins indicates that cyclin A, B and E expression is similarly regulated, while other factors regulate cyclin D1 expression. The results suggest that the combined immunoreactivity of cyclins A, B1, D and E might be a useful prognostic factor in breast cancer.

Matrix metalloproteinase (MMP)-1, -9 and -13 as prognostic factors in salivary gland cancer.

CONCLUSIONS: In the current study matrix metalloproteinases (MMPs)-9 and -13 were associated with the prognosis in salivary gland cancer (SGC), indicating that they contribute to the progression and invasion of these malignant tumours. OBJECTIVES: Elevated levels of certain MMPs have been detected in various advanced human cancer types. The purpose of the study was to analyse the expression of MMP-1, -9 and -13 in SGC by immunohistochemistry and correlate the results to the clinical data and 10-year survival of SGC patients in a nationwide material. MATERIALS AND METHODS: Immunohistochemistry for MMP-1, -9 and -13 was performed in series of 103 paraffin-embedded sections of SGC. RESULTS: High MMP-13 staining intensity predicted poor survival (3 vs 1; p=0.08) in the whole material studied. High MMP-13 intensity (2+3 vs 1; p=0.05), percentage (2 vs 1; p=0.03) and index (3 vs 1; p=0.02) were associated with poor survival in acinic cell carcinoma. However, high MMP-9 index in adenoid cystic carcinoma (p=0.06) and the percentage of positively staining cells in salivary duct carcinoma patients (p=0.05) was associated with poor survival. High MMP-1 staining intensity (2 vs 0; p=0.06) and index (% x intensity); (2 vs 1, p=0.04) were associated with better overall survival in the whole material.

Prognostic significance of matrix metalloproteinases-1, -2, -7 and -13 and tissue inhibitors of metalloproteinases-1, -2, -3 and -4 in colorectal cancer.

Strong expression of many matrix metalloproteinases (MMPs) has been related to poor survival of colorectal cancer (CRC) patients. The expression of tissue inhibitors of metalloproteinases (TIMPs) has been associated with both a beneficial and a poor outcome and there is thus a need to further clarify the significance of MMPs and TIMPs in CRC. The prognostic significance of 4 MMPs and TIMPs in CRC was evaluated. Formalin-fixed, paraffin-embedded tissue arrayed samples of 351 patients with primary colon or rectal cancer of Dukes' stages A-D were selected for immunohistochemical staining of MMP-1, -2, -7 and -13, and TIMP-1, -2, -3 and -4. High expression of MMP-2 in the malignant epithelium as well as in the surrounding stroma was associated with reduced survival of colon cancer patients. Strong epithelial and stromal cytoplasmic staining of TIMP-3 was associated with a longer survival in rectal cancer patients, and here the interobserver variation for evaluating the degree of staining was lower than for epithelial staining. Strong stromal cytoplasmic staining of TIMP-4 predicted longer survival of rectal cancer patients. Multivariate analysis showed that stromal cytoplasmic TIMP-3 staining was the only marker of independent prognostic value. MMP-2 might be a useful prognostic marker in colon cancer, and TIMP-3 and TIMP-4 in rectal cancer, but the findings associated with stromal staining should be interpreted with some caution. Different biologic behavior or different genetic development may explain the differences between colon and rectal cancers regarding the expression of MMP-2, TIMP-3 and TIMP-4.

The significance of tumor markers for proliferation and apoptosis in predicting survival in colorectal cancer.

PURPOSE: Clinicopathologic staging is even today the best prognostic factor in both colon and rectal cancers. There is still considerable variation in survival within the stages. To find other prognostic indicators we investigated six biologic markers associated with apoptosis and cell proliferation. METHODS: Formalin-fixed, paraffin-embedded tissue samples of 363 patients with primary colon or rectal cancer of Dukes Stages A to D were chosen for immunohistochemical staining of five tumor markers: bcl-2, p53, Ki-67, cyclin D1, and carcinoembryonic antigen. Also, the number of apoptotic cells was studied by the terminal deoxynucleotidyl transferase-mediated D: -UTP nick end labeling method in 347 cases. The study was done on specially prepared tissue arrays. RESULTS: In rectal cancer, patients with a Ki-67 labeling index of 5 percent or higher had a better prognosis than those with a lower index. Also, positive cytoplasmic p53 expression predicted a favorable outcome in rectal cancer. In colon cancer, positive nuclear staining of cyclin D1 reflected better survival. Weak and moderate staining of carcinoembryonic antigen correlated with better prognosis than strong staining, but negative staining predicted poor outcome. High apoptotic index of 100 or higher correlated with poor prognosis in colon cancer. However, in rectal cancer, the trend was the opposite. Bcl-2 staining tended to be more intense in samples of patients living 5 years or longer compared with those with worse prognosis. CONCLUSIONS: Colon cancer and rectal cancer seem to have different biologic behavior, at least with respect to apoptosis, cytoplasmic p53 expression, and perhaps Ki-67 and carcinoembryonic antigen. Further studies are needed to clarify the significance of these factors.

Toxicity of antiestrogens.

The object of this article is to review briefly the preclinical and clinical safety of some antiestrogens. Tamoxifen, toremifene, droloxifene, and idoxifene are polyphenylethylene antiestrogens, whereas the pure antiestrogen, ICI 182,780 or faslodex, as well as raloxifene, is of a different structure. Tamoxifen has been shown to be genotoxic in several studies. It induces unscheduled DNA synthesis in rat hepatocytes and micronuclei in MCL-5 a cells in vitro. Tamoxifen also induces aneuploidy in rat liver in vivo and chromosome aberrations and micronuclei in mouse bone marrow. Toremifene has also shown to be genotoxic, but to a far lower extent, by inducing micronuclei in MCL-5 a cells in vitro and by inducing aneuploidy in rat liver in vivo. Tamoxifen has been shown to be hepatocarcinogenic in the rat in at least four independent long-term studies. The initiation of tumors in the rat is the result of metabolic activation by cytochrome P450 isoenzymes to an electrophile(s) that binds irreversibly to DNA. The other antiestrogens have not been shown to be carcinogenic in rodents. In several independent clinical studies, the risk of endometrial cancer has increased among tamoxifen-treated women. After reviewing the available data, the International Agency for Research on Cancer concluded that there was sufficient evidence to show that tamoxifen is a class I human carcinogen. The increased risk for endometrial cancer occurs predominantly among women who are 50 years old or older and who have been treated with tamoxifen. It is not yet clear whether the uterine tumor formation is a result of genetic mechanisms, analogous to those seen in the rat liver or due to the estrogen agonist action of tamoxifen. However, the other antiestrogens with a more or less similar intrinsic estrogenic potential have not been shown to be carcinogenic in humans.