Prognosis In Invasive Ductal Carcinoma Research Articles

Expression of e-cadherin, n-cadherin and snail and their correlation with clinicopathological variants: an immunohistochemical study of 132 invasive ductal breast carcinomas in Egypt

OBJECTIVE:To evaluate the expression of the cell adhesion molecules E-cadherin and N-cadherin and the transcription factor Snail in invasive ductal breast carcinomas and to determine their relationships with clinicopathological features.METHODS:Immunohistochemistry was used to examine E-cadherin, N-cadherin, and Snail protein expression in 132 invasive breast carcinomas.RESULTS:The expression of E-cadherin was decreased (negative or weak) in 37.1% of invasive carcinomas, while N-cadherin and Snail overexpression were detected in 51.9% and 40.9% of carcinomas, respectively. Low E-cadherin expression was significantly correlated with poorly differentiated carcinoma (53.1%), positive node status (80.9%), poor Nottingham Prognostic Index (64.7%), and the presence of estrogen and progesterone receptors. Overexpression of N-cadherin and Snail were also significantly correlated with poorly differentiated carcinoma, positive node status, and poor Nottingham Prognostic Index but were correlated with the absence of hormone receptors. Loss of E-cadherin immunoexpression was strongly associated with the presence of membranous N-cadherin (87.8%) and nuclear Snail (69.4%).CONCLUSION:Loss of E-cadherin and overexpression of N-cadherin and Snail in breast carcinomas may play a central role in the development of invasive ductal breast carcinoma. These biomarkers may provide a valuable reference for the study of invasive ductal carcinoma progression and to characterize the biological behavior of the tumor. In the future, increased N-cadherin and decreased E-cadherin expression may be used as indicators of the progression and prognosis of invasive ductal carcinoma.

X-linked inhibitor of apoptosis positive nuclear labeling: a new independent prognostic biomarker of breast invasive ductal carcinoma

BackgroundIt's well recognized that X-linked inhibitor of apoptosis (XIAP) was the most potent caspase inhibitor and second mitochondria-derived activator of caspase (Smac) was the antagonist of XIAP. Experiments in vitro identified that down regulation of XIAP expression or applying Smac mimics could sensitize breast cancer cells to chemotherapeutics and promote apoptosis. However, expression status and biologic or prognostic significance of XIAP/Smac in breast invasive ductal carcinoma (IDC) were not clear. The present study aimed to investigate relationship among expression status of XIAP/Smac, apoptosis index (AI), clinicopathologic parameters and prognosis in IDC.MethodsImmunohistochemistry and TUNEL experiment were performed to detect expression of XIAP, Smac, ER, PR, HER2 and AI in 102 cases of paraffin-embedded IDC samples respectively. Expression of XIAP/Smac were also detected in limited 8 cases of fresh IDC specimens with Western blot.ResultsPositive ratio and immunoscore of XIAP was markedly higher than Smac in IDC (P < 0.0001). It was noteworthy that 44 cases of IDC were positive in nuclear for XIAP, but none was for Smac. Expression status of Smac was more prevalent in HER2 positive group than negative group (P < 0.0001) and AI was positively correlated with HER2 protein expression (rs = 0.265, P = 0.017). The present study first revealed that XIAP positive nuclear labeling (XIAP-N), but not cytoplasmic staining (XIAP-C), was the apoptotic marker correlated significantly with patients' shortened overall survival (P = 0.039). Survival analysis demonstrated that XIAP-N was a new independent prognostic factor except for patient age and lymph node status.ConclusionDisturbed balance of expression between XIAP and Smac probably contributed to carcinogenesis and XIAP positive nuclear labeling was a new independent prognostic biomarker of breast IDC.

The impact of caveolin protein expression in tumor stroma on prognosis of breast cancer

We aimed to investigate the expression of caveolin-1, -2, -3, and platelet-derived growth factor (PDGF) β receptor in breast cancer cells and stroma by immunohistochemistry and to analyze their implications. The expression rates of stromal caveolin-2 and PDGF β receptor increased as the tumor progressed from ductal carcinoma in situ to microinvasive ductal carcinoma, intraductal component of invasive ductal carcinoma (IDC), and IDC (p<0.001). The expression loss of caveolin-1 in tumor stroma of IDC correlated with high tumor stage (p<0.001), high nodal stage (p=0.011), high cancer stage (p=0.005), estrogen receptor negativity (p=0.003), and tumor recurrence (p=0.003). In addition, the expression loss of caveolin-1 in tumor stroma was correlated with a shorter disease-free survival and an overall survival (p<0.001). In conclusion, the loss of stromal caveolin-1 is related to poor prognosis in IDC.

40. Cytoplasmic localisation of β catenin is a marker of poor outcome in breast cancer patients

β catenin is involved in cell adhesion via catenin-cadherin complexes and as a transcriptional regulator in the Wnt signaling pathway. Its deregulation is important in the genesis of a number of human malignancies, particularly colorectal cancer. A range of studies have been undertaken in breast cancer but there is a lack of consensus on associations reported between β catenin expression, clinicopathological parameters and disease outcome. We undertook an immunohistochemical study measuring the levels and subcellular localisation of β catenin in 292 invasive ductal breast cancers with known treatment and outcome. Forty percent of tumours were >20mm, 45% were grade >2, 43% were lymph node positive, 68% were ER positive, 57% were PR positive and 18% were HER2 FISH positive. The median age was 54 years and patients were treated with endocrine therapy (49%), chemotherapy (38%) or both (24%). Cases were prospectively followed-up for a median of 64 months and outcome events measured were: recurrence (local or distant) (25%), metastasis (23%) and all deaths were recorded but only breast cancer-related deaths (18%) were considered for survival analyses. β catenin expression was found predominantly in the membrane and cytoplasm of cells, with only 3 cases showing detectable nuclear expression, hence we described a histoscore value for the cytoplasmic and the membrane expression independently, and then derived a third histoscore which represents both categories of expression in one continuous variable (membrane minus cytoplasmic histoscore) as a measure of subcellular localisation, the ‘membrane to cytoplasmic score' (MTC). When this continuous value is positive the tissue shows predominantly membranous expression and when the value is negative the tissue shows mainly cytoplasmic expression. No association with outcome was observed for cytoplasmic or membrane expression alone, however a shift from membrane to cytoplasm expression calculated using the MTC score was associated with worse outcome in univariate analysis (<i>p</i>=0.004), and approached significance in a multivariate analysis model that included lymph node, PR and HER2 status (<i>p</i>=0.054). The MTC score was then used for further statistical analyses due to the importance of both subcellular location and levels of expression of β catenin. An association was identified between a shift to high cytoplasmic expression (low MTC score) and high tumour grade (<i>p</i>=0.004), positive Ki67(<i>p</i>=0.0001), negative ER (<i>p</i>=0.005), positive HER2(<i>p</i>=0.01) status and an active PI3K pathway (<i>p</i>=0.005), measured as PIK3CA mutations (<i>p</i>=0.05) or PTEN loss (<i>p</i>=0.05). A shift to low cytoplasmic expression (high MTC score) was associated with the Luminal A subtype (<i>p</i>=0.004). A shift in subcellular localisation of β catenin is associated with alterations in cell cycle regulatory proteins and signalling transduction pathways, in clinical breast cancer samples. Furthermore, cytoplasmic accumulation of β catenin is associated with a poor prognosis in invasive ductal carcinoma. It is possible that in a larger cohort, independence may be established and further efforts are needed to validate these findings in other breast cancer cohorts.

Abstract S6-8: Hedgehog Ligand Overexpression Predicts Poor Outcome in Breast Cancer and Is a Potential Therapeutic Target for Metastatic Disease

Abstract Background: The Hh signalling pathway plays an important role in a number of malignancies, and accumulating data suggest it contributes significantly to the development and progression of breast cancer. However, there is scant data regarding the clinical significance of its dysregulation in breast cancer and its functional effects in breast cancer models. Methods: We investigated the clinical significance of the expression of Hh pathway components using immunohistochemistry in a well-characterised cohort of 279 patients with invasive ductal carcinoma (IDC) to determine its prognostic significance. A murine mammary cancer allograft model based on the M6 cell line, derived from the C3/SV40T transgenic mouse was used to determine the effects of Shh overexpression on tumour growth in vitro and in vivo. Finally, the therapeutic potential of Hh blockade using the monoclonal antibody 5E1 was also explored in two mouse mammary carcinoma models; the M6 allograft model and the highly metastatic 4T1 cell line in immunocompetent Balb/C mice. Results: High Hh ligand expression was observed in 34% of IDC and was associated with increased risk of breast cancer recurrence (HR 1.95, p=0.0004) and breast cancer specific death (HR 2.3, p=0.0002). High Hh expression was also associated with the basal-like subtype (p=0.004). Overexpression of Hh ligand in M6 cells transplanted to fat pads of immunodeficient mice resulted in a 4 fold increase in tumor volume (p= 0.006) and was associated with upregulation of the canonical Hh target genes Gli1 and HHIP. However, Shh promoted peritumoral lymphatic invasion in 4/5 mice compared to only 1/5 in the vector control group and was associated with a significantly shorter time to the development of metastatic disease (p=0.0004). M6 cells overexpressing Hh ligand showed no change in proliferation, self renewal or migration in vitro, suggesting a requirement for stromal interaction in Hh-dependent tumor promotion. Finally, we found that the enhanced tumor growth conferred by Hh ligand overexpression could be blocked by administration of a neutralizing antibody to Hh ligand. Furthermore, using a metastatic mammary carcinoma isograft model (4T1) that endogenously expresses Hh ligand, we demonstrate that Hh blockade resulted in smaller lung metastatic deposits (p=0.045). Conclusions: We found that high Hh ligand is associated with a poor prognosis in IDC. Overexpression of Hh ligand promotes murine mammary tumor growth, which requires stromal interaction, as well as lymphatic vessel invasion. Finally, we demonstrate that blockade of the Hh ligand is a potential therapy in metastatic breast carcinoma. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr S6-8.

Should Histologic Grade Be Incorporated into the TNM Classification System for Small (T1, T2) Node-Negative Breast Adenocarcinomas?

Prognosis of invasive ductal carcinoma (IDC) strongly correlates with tumor grade as determined by Nottingham combined histologic grade. While reporting grade as low grade/favorable (G1), intermediate grade/moderately favorable (G2), and high grade/unfavorable (G3) is recommended by American Joint Committee on Cancer (AJCC) staging system, existing TNM (Primary Tumor/Regional Lymph Nodes/Distant Metastasis) classification does not directly incorporate these data. For large tumors (T3, T4), significance of histologic grade may be clinically moot as those are nearly always candidates for adjuvant therapy. However, for small (T1, T2) node-negative (N0) tumors, grade may be clinically relevant in influencing treatment decisions, but data on outcomes are sparse and controversial. This retrospective study analyzes clinical outcome in patients with small N0 IDC on the basis of tumor grade. Our results suggest that the grade does not impact clinical outcome in T1N0 tumors. In T2N0 tumors, however, it might be prognostically significant and relevant in influencing decisions regarding the need for additional adjuvant therapy and optimal management.

29. c-MYC gene amplification is associated with a poor prognosis in invasive ductal carcinoma

Background c-MYC is a proto-oncogene located on 8q24 with well-established critical roles in the development and progression of breast cancer. However, there is relatively little information available on the frequency of its amplification in breast cancer, with widely varying estimates in the literature and only two studies using the gold-standard of fluorescent in situ hybridisation (FISH). Moreover, there is little consensus as to a clinically relevant c-MYC/CEP8 (centromeric region of chromosome 8) ratio in determining patient outcome or response to therapy. Methods We investigated the frequency of c-MYC gene amplification using FISH in formalin fixed, paraffin processed tissues in 16 tissue microarrays from a cohort of 262 patients with operable invasive ductal carcinoma. C-MYC gene and CEP8 region copy number were determined in 20 representative tumour nuclei and a ratio c-MYC/CEP8 calculated. Two different ratios were tested using Kaplan-Meier survival analysis for their prognostic strength; 2.2 as used to determine HER2 gene amplification, and a newly described ratio of 1.3 which co-segregated with outcome. Chi squared analysis was then used to investigate the clinicopathological associations of c-MYC gene amplification Results A c-MYC/CEP8 ratio of 1.3 identified a poor prognostic group in the cohort (HR 2.28, p = 14; 0.006), while a ratio of 2.2 showed only a trend towards significance (p = 14; 0.087). Using a ratio of 1.3, 22.1% of cancers showed c-MYC gene amplification, while 61% were diploid and 16.9% were polysomic. c-MYC gene amplification was associated with larger, grade 3, ER negative, Ki67 high tumours (all p 5 0.05). Furthermore there was a highly significant association with the basal-like phenotype (p = 14; 0.004). Conclusion We have validated that a c-MYC/CEP8 ratio of 1.3 is able to identify a poor prognosis group of breast cancer patients. c-MYC gene amplification is associated with a number of high risk clinicopathological variables.

S100A8 and S100A9 Overexpression Is Associated with Poor Pathological Parameters in Invasive Ductal Carcinoma of the Breast

S100 protein A8 and A9 naturally form a stable heterocomplex. Recently, we have proved that S100A9 overexpression in various adenocarcinomas is associated with poor tumor differentiation. In this study, we examined the relationship between the expression of each protein and the pathological parameters that reflect the aggressiveness of carcinoma, in invasive ductal carcinoma (IDC) of the breast. Serial paraffin-embedded tissue sections from 101 IDC cases were immunostained with respective monoclonal antibodies, and the results were as follows: 1) A positive correlation of immunoreactivity between S100A8 and S100A9 was noticed (r=0.873 and P<0.0001); 2) The percentage of S100A9-positive tumor cells was higher than that of S100A8-positive tumor cells (P<0.001), and S100A8 alone was not detected in any case; 3) Overlap between S100A8 and S100A9 staining patterns was found in the corresponding tissue areas, but S100A9 positivity was also observed in S100A8-negative tumor cells; 4) The immunopositivity for each protein also correlated with the mitotic activity, MIB-1 index, HER2 overexpression, node metastasis, and poor pT categories and pStage (P<0.05); 5) Co-expression of both proteins was associated with poor tumor differentiation, vessel invasion, node metastasis, and poor pStage (P<0.05). Furthermore, co-expression of the proteins was also observed in MCF-7 cells, and it was suggested that the immunolocalization is related with cell cycle. Our conclusions are as follows: 1) It is suggested that S100A8 is S100A9-dependently expressed and acquires the protein stability by S100A8/A9 heterocomplex formation; 2) S100A8 and S100A9 overexpression should be considered marker of poor prognosis in IDC.

Expression of MUC5AC and MUC6 in invasive ductal carcinoma of the pancreas and relationship with prognosis.

MUC5AC and MUC6 are two major types of mucin that are abundantly present in the stomach; both of them form a gel of high viscosity that provides protection and lubrication. Expressions of MUC5AC and MUC6 are seen in pancreatic neoplasms, whereas the relationships between MUC5AC/MUC6 expression and clinicopathological factors and patient prognosis in invasive ductal carcinoma (IDC) of the pancreas have not been investigated. The aim of this study was to investigate MUC5AC and MUC6 expressions in IDC with special reference to clinicopathological factors and patient prognosis. Tissue samples were taken from 33 patients with IDC of the pancreas after radical surgical treatment. MUC5AC and MUC6 expressions were examined immunohistochemically. The expressions of MUC5AC and MUC6 were observed in the cytoplasm of the tumor cells. MUC5AC and MUC6 immunoreactivities in the cancer tissues were found in 21 (63.6%) and 15 (45.5%) of 33 cases of IDC of the pancreas, respectively. MUC5AC-negative expression was associated significantly with lymphatic invasion, venous invasion, lymph node metastasis, and MUC5AC-positive patients showed significant better survival than those MUC5AC-negative patients. MUC6 expression was significantly related to tumor location, whereas MUC6 expression did not show significant relationship with patient survival. The results indicate that MUC5AC expression plays an important role in impacting tumor progression in IDC of the pancreas. MUC5AC expression is a benefit to better survival of patients with IDC of the pancreas. MUC6 expression is not involved in tumor progression in IDC of the pancreas.