Invasive Ductal Breast Cancer Patients Research Articles

18F-FDG Uptake by Metastatic Axillary Lymph Nodes on Pretreatment PET/CT as a Prognostic Factor for Recurrence in Patients with Invasive Ductal Breast Cancer

This study assessed the maximum standardized uptake value of metastatic axillary lymph nodes in patients with invasive ductal breast cancer (IDC) to determine the pretreatment prognostic value of (18)F-FDG PET/CT for disease-free survival (DFS). Sixty-five female IDC patients who had undergone pretreatment (18)F-FDG PET/CT and had pathologically confirmed axillary lymph node involvement without distant metastasis were enrolled. All patients showed complete remission after first-line treatment. To obtain nodal SUVmax, a transaxial image representing the highest (18)F-FDG uptake was carefully selected and a region of interest was manually drawn on the (18)F-FDG-accumulating lesion. Clinicopathologic parameters such as age, TNM stage, estrogen receptor status, progesterone receptor status, human epidermal growth factor receptor 2 status, and primary-tumor and nodal SUVmax on PET were analyzed for their usefulness in predicting recurrence. Combinatorial effects and interactions between variables that were significant by univariate analysis were examined using multivariate Cox proportional-hazards models. Twelve of 65 patients (18.5%) experienced recurrence during follow-up (median follow-up, 36 mo; range, 21-57 mo). Nodal SUVmax was significantly higher in patients with recurrence than in those who were disease-free (recurrence group: 5.2 ± 2.3, vs. disease-free group: 1.9 ± 1.9, P < 0.0001). A receiver-operating-characteristic curve demonstrated a nodal SUVmax of 2.8 (sensitivity, 91.7%; specificity, 86.8%; area under the curve, 0.890) to be the optimal cutoff for predicting DFS. Univariate analysis revealed that T stage, N stage, estrogen receptor status, and primary-tumor and nodal SUVmax correlated significantly with DFS. Among these 5 variables, only nodal SUVmax was found to be a single determinant of DFS by multivariate analysis (hazard ratio, 31.54; 95% confidence interval, 2.66-373.39; P = 0.0065). Nodal SUVmax on pretreatment (18)F-FDG PET/CT may be an independent prognostic factor for disease recurrence in patients with IDC.

Increased expression of Rab25 in breast cancer correlates with lymphatic metastasis

Breast cancer is the most common cancer in women worldwide. Studies have suggested that Ras-related protein 25 (Rab25), a member of Rab small GTPase family, is involved in the pathogenesis of breast cancer. In this study, we investigated whether the expression of Rab25 correlated with lymphatic metastasis in breast cancer and whether the expression of Rab25 was positively correlated with oestrogen receptor (ER) and progesterone receptor (PR) expression in breast cancer. Breast cancer tissues from 42 invasive ductal breast cancer patients with or without lymphatic metastasis were collected and the levels of Rab25 mRNA and protein measured by quantitative real-time PCR and immunohistochemistry, respectively. The mRNA level of Rab25 was significantly increased in invasive ductal breast cancer with lymphatic metastasis compared to that in invasive ductal breast cancer without lymphatic metastasis. Immunohistochemical analysis demonstrated that Rab25 and vascular endothelial growth factor (VEGF) were highly expressed in invasive ductal breast cancer with lymphatic metastasis regardless of whether the cancer is ER and PR positive or negative. Higher expression of Rab25 positively correlated with VEGF expression. However, the expressions of Rab25 in ER and PR-positive cancers were much higher than ER and PR-negative cancers regardless of whether lymphatic metastasis occurred. These data suggest that higher level of Rab25 was associated with lymphatic metastasis, specifically in ER and PR-positive breast cancer. The better understanding of the mechanism of Rab25 may provide a basis for the development of a novel therapeutic target in breast cancer.

Microcephalin is a new novel prognostic indicator in breast cancer associated with BRCA1 inactivation

The authors have investigated the expression of the microcephalin (MCPH1) protein to evaluate its prognostic importance in breast cancer. Microcephalin is a damage response protein involved in the regulation of BRCA1 and BRCA2. BRCA1 mutations are often associated with basal-like breast cancer, which are also often negative for oestrogen receptor (ER), progesterone receptor (PR) and HER2. MCPH1 immunohistochemistry was performed on 319 breast cancers prepared as tissue microarray and correlated with pathology, survival, ER, PR, HER2, EGFR, CK5/6, CK14 and BRCA1 expression. After performing continuous data analysis, mean microcephalin expression decreased with increasing grade (P < 0.006). Mean microcephalin expression was lower in ER/PR negative (P < 0.001) and triple negative cancers (P < 0.004). Conversely, an association with HER2-positive cancers was also identified (P < 0.034). Reduced microcephalin also correlated with reduced nuclear BRCA1 staining (P < 0.001). No association was identified with basal markers. After dichotomising the data into low and high microcephalin expression, reduced expression was identified in 29% (93/319) of breast cancers. An association with low expression was identified in invasive ductal carcinomas with breast cancer-specific survival (BCSS) (P = 0.052). Multivariate analysis of ductal carcinomas showed that microcephalin, together with lymph node involvement and tumour size were independent predictors of BCSS (P = 0.037). Microcephalin expression is reduced in 29% of breast cancers, particularly in higher grade tumours and BRCA1-negative cases. Microcephalin is an independent predictor of BCSS in invasive ductal breast cancer patients and may prove to be a useful biomarker for the identification of aggressive breast cancers.

Reproductive Factors, Steroid Receptor Status, and Tumour Markers of HER2-Positive Breast Cancer in Northern China

Background: Although China has a breast cancer incidence that surpasses all other cancer registries, there have been few reports to evaluate the relationships of reproductive history, steroid receptor status, and tumour markers with HER2 status of breast cancer. Patients and Methods: This project included 274 primary invasive ductal breast cancer patients. Information concerning the reproductive factors and tumour characteristics of the patients had previously been collected. HER2 and steroid receptor status were detected in tumour tissues. Serum CEA, CA15-3, and CA125 levels were analyzed for all patients. Results: Younger age at menarche was observed in patients with HER2-positive than in those with HER2negative status (p = 0.03). Statistically significant differences were found between the HER2-positive group and HER2-negative group for estrogen receptor (ER) and progesterone receptor (PR) status (p < 0.01). Age of onset, other reproductive factors, tumour characteristics, and serum tumour marker level were not significantly different between those patients with HER2-positive and those with HER2-negative status. Conclusion: We confirm that age at menarche may plausibly be differentially correlated with the risk of HER2-positive invasive ductal breast cancer because it is presumed to impact exposure to endogenous sex hormones. HER2 is inversely related to ER and PR in invasive ductal breast cancer patients of Northern China.

Prognostic differences of World Health Organization–assessed mitotic activity index and mitotic impression by quick scanning in invasive ductal breast cancer patients younger than 55 years

Prognostic differences of World Health Organization–assessed mitotic activity index and mitotic impression by quick scanning in invasive ductal breast cancer patients younger than 55 years

Tumor necrosis factor gene polymorphisms in breast cancer patients

Analysis of microsatellite TNFalpha marker and (-308(G/A) polymorphisms in promoter of TNFalpha gene was conducted in 167 patients with various types of sporadic breast cancer (BC) as well as in 139 healthy Russian donors. It was shown that frequency of allele 7 in TNFalpha microsatellite marker was significantly higher in BC patients than in healthy donors (17.9% versus 10.4%; P = 0.02) mainly due to the patients with invasive ductal BC (19.2% versus 10.4%; P = 0.008). The TNFalpha allele 9 was observed significantly more frequently in patients with invasive-ductal cancer (6.4% versus 1%; P= 0.01). The studies of-308(G/A)TNFalpha polymorphism in BC patients and healthy donors have shown no differences in the distribution frequency of highly secreted allele (-308A)TNFalpha. However, invasive lobular BC patients carrying (-308AG)TNFalpha genotype were observed significantly more frequently than invasive-ductal BC patients carrying the same allele (34.0 versus 17.3%; P = 0.034). Thus it has been shown for the first time that invasive-ductal and invasive-lobular BC patients differ in distribution of TNFalpha and -308(G/A)TNFalpha alleles.

Prognostic differences of World Health Organization–assessed mitotic activity index and mitotic impression by quick scanning in invasive ductal breast cancer patients younger than 55 years

The proliferation marker mitotic activity index is the strongest prognostic indicator in lymph node-negative breast cancer. The World Health Organization (WHO) 2003-defined procedure for determining WHO-mitotic activity index is often replaced by a quick scan mitotic impression. We evaluated the prognostic consequences of this practice in 433 T(1-3)N(0)M(0) lymph node-negative invasive ductal type breast cancers with long-term follow-up (median, 112 months; range, 12-187 months). Twenty-seven percent of the studied cases developed distant metastases, and 25% died of disease. Agreement between WHO-mitotic activity index (0-5 = 1, 6-10 = 2, >10 = 3) and mitotic impression (1, 2, 3) categories was 66% (kappa = 0.41), including 85% for category 1, 26% for category 2, and 52% for category 3. The WHO-mitotic activity index was a much stronger prognosticator than the mitotic impression, and the 10-year survival rates of the same categories (eg, mitotic activity index and mitotic impression category both 2) differed greatly. When grade was assessed by combining WHO-mitotic activity index or mitotic impression with the same values for tubular formation and nuclear atypia, grades disagreed in 18% of the cases. Deviation from the formal WHO-mitotic activity index assessment guidelines in breast cancer often results in erroneous prognosis estimations with therapeutic consequences and may explain why the prognostic value of proliferative activity in breast cancer is not always confirmed.

Whole Genome Amplification of Plasma-Circulating DNA Enables Expanded Screening for Allelic Imbalance in Plasma

Apoptotic and necrotic tumor cells release DNA into plasma, providing an accessible tumor biomarker. Tumor-released plasma-circulating DNA can be screened for tumor-specific genetic changes, including mutation, methylation, or allelic imbalance. However, technical problems relating to the quantity and quality of DNA collected from plasma hinder downstream genetic screening and reduce biomarker detection sensitivity. Here, we present a new methodology, blunt-end ligation-mediated whole genome amplification (BL-WGA), that efficiently amplifies small apoptotic fragments (<200 bp) as well as intermediate and large necrotic fragments (>5 kb) and enables reliable high-throughput analysis of plasma-circulating DNA. In a single-tube reaction, purified double-stranded DNA was blunted with T4 DNA polymerase, self-ligated or cross-ligated with T4 DNA ligase and amplified via random primer-initiated multiple displacement amplification. Using plasma DNA from breast cancer patients and normal controls, we demonstrate that BL-WGA amplified the plasma-circulating genome by approximately 1000-fold. Of 25 informative polymorphic sites screened via polymerase chain reaction-denaturating high-performance liquid chromatography, 24 (95%) were correctly determined by BL-WGA to be allelic retention or imbalance compared to 44% by multiple displacement amplification. By enabling target magnification and application of high-throughput genome analysis, BL-WGA improves sensitivity for detection of circulating tumor-specific biomarkers from bodily fluids or for recovery of nucleic acids from suboptimally stored specimens.

Protein Kinase Cε Is a Predictive Biomarker of Aggressive Breast Cancer and a Validated Target for RNA Interference Anticancer Therapy

Tumor metastasis is the major cause of morbidity and mortality in patients with breast cancer. It is critical to identify metastasis enabling genes and understand how they are responsible for inducing specific aspects of the metastatic phenotype to allow for improved clinical detection and management. Protein kinase C epsilon (PKC epsilon), a member of a family of serine/threonine protein kinases, is a transforming oncogene that has been reported to be involved in cell invasion and motility. In this study, we investigated the role of PKC epsilon in breast cancer development and progression. High-density tissue microarray analysis showed that PKC epsilon protein was detected in 73.6% (106 of 144) of primary tumors from invasive ductal breast cancer patients. Increasing PKC epsilon staining intensity was associated with high histologic grade (P = 0.0206), positive Her2/neu receptor status (P = 0.0419), and negative estrogen (P = 0.0026) and progesterone receptor status (P = 0.0008). Kaplan-Meier analyses showed that PKC epsilon was significantly associated with poorer disease-free and overall survival (log-rank, P = 0.0478 and P = 0.0414, respectively). RNA interference of PKC epsilon in MDA-MB231 cells, an aggressive breast cancer cell line with elevated PKC epsilon levels, resulted in a cell phenotype that was significantly less proliferative, invasive, and motile than the parental or the control RNA interference transfectants. Moreover, in vivo tumor growth of small interfering RNA-PKC epsilon MDA-MB231 clones was retarded by a striking 87% (P < 0.05) and incidence of lung metastases was inhibited by 83% (P < 0.02). PKC epsilon-deficient clones were found to have lower RhoC GTPase protein levels and activation. Taken together, these results revealed that PKC epsilon plays a critical and causative role in promoting an aggressive metastatic breast cancer phenotype and as a target for anticancer therapy.

Multivariate prognostic evaluation of the mitotic activity index and fibrotic focus in node-negative invasive breast cancers

We validated with univariate and multivariate (Cox) analysis, the prognostic value of the mitotic activity index (MAI), the fibrotic focus (FF) and other prognosticators in 448 patients with lymph node-negative (LN−) invasive breast cancer <55 years without adjuvant systemic treatment (72.5 months median follow-up, range 4–119). Of these patients, 24.8% developed distant and 1.6% loco-regional recurrence. FF showed excellent inter-observer reproducibility ( κ = 0.93). Strong prognosticators were MAI, grade, nuclear atypia, FF and the St. Gallen criterion (SG). The subgroup with excellent survival selected by SG was only 16% of all patients, implying over-treatment of more than 70% of all LN− patients when using SG as adjuvant therapy selection criterion. If MAI <10, 13% showed distant metastases, contrasting with 41% if MAI ⩾10. FF was prognostic in the ductal and mixed ductal cancers, but not in the lobular and other subtype cancers. Patients with invasive (mixed) ductal cancers with FF absent, FF < 1/3 or FF > 1/3 of the tumour area, had distant metastasis rates of 17%, 35% and 48%; in MAI < 10 and FF absent, FF < 1/3 or FF > 1/3, metastasis rates were 11%, 13% and 42% and if MAI ⩾ 10, metastasis rates were 31%, 48% and 50%, respectively. In the 12 patients with MAI < 10 and a large FF > 1/3, event-free survival was similar to patients with MAI ⩾ 10. With multiple regression MAI < 10 versus ⩾10 is the strongest prognosticator (also stronger than the SG). The FF may be important as it has additional prognostic value to the MAI in the small subgroup of invasive ductal or mixed-ductal breast cancer patients with combined MAI < 10 and an FF > 1/3 of the tumour area.

Tumor-derived aberrant methylation in plasma of invasive ductal breast cancer patients: Clinical implications

Progressive p16 methylation has been associated with metastasis and invasive phenotypes in many cancers. Loss of E-cadherin (CDH1) function contributes to breast cancer progression by promoting cell proliferation, invasion and metastasis. Using methylation-specific PCR, aberrant hypermethylation of p16 and CDH1 in tumor and plasma was analyzed and correlated with levels of serum protein markers, carcinoembryonic antigen (CEA) and carcinoma antigen 15-3 (CA15.3), in 36 patients with invasive ductal breast cancer. Aberrant p16 methylation was found in 11% (4/36) of primary tumors and 8% (3/36) of plasma samples. Aberrant CDH1 methylation was detected in 25% (9/36) of primary tumors and 20% (7/36) of plasma samples. p16 and/or CDH1 hypermethylation was found in 31% (11/36) of primary breast carcinomas and 82% (9/11) of breast cancer patients with tumoral methylation showing identical epigenetic changes in plasma. The 25 patients without tumoral methylation did not show epigenetic changes in the plasma. Tumoral p16 methylation was significantly associated with advanced tumor stage (p=0.028; Fisher's exact test), tumor size (p=0.017) and nodal metastasis (p=0.002). However, p16 methylation in plasma was only associated with nodal metastasis (p=0.012). Altogether, aberrant p16 methylation in plasma and elevated serum CEA level were associated with advanced tumor stage (p=0.033), tumor size (p=0.022) and extensive nodal metastasis (p=0.003). With clinical implications, p16 hypermethylation in plasma and/or raised serum CEA levels may prove useful as diagnostic and prognostic markers for breast cancer.

Transcriptional and translational modulation of KAI1 expression in ductal carcinoma of the breast and the prognostic significance

KAI1, also known as CD82, has been shown to have a potential impact on the invasiveness of cancer cells. In the present study, expression pattern of KAI1, both at transcription and translation levels and the potential clinical value of the expression were explored in a cohort of normal and ductal mammary cancer tissues (n=71). A marked reduction of KAI1 transcript was observed in invasive ductal breast tumours as compared to normal tissues. Expression of KAI1 protein was higher in normal tissues as compared to tumour samples. Though no significant difference of KAI1 expression between different grades of tumour was observed (p=0.064), significant correlation of TNM staging with KAI1 expression has been observed in invasive ductal breast cancer patients (p=0.045). Additionally, it was also observed that patients showing higher expression of KAI1 had a longer 10-year survival rate as compared to a low level or completely negative expression KAI1 (p=0.0136). KAI1 inverse correlation with tumour progression may be used as a strong prognostic marker.

P53 overexpression is a prognostic indicator in poorly differentiated, node negative and T1/T2 invasive ductal breast cancer patients

The present study evaluates the prognostic value of p53 protein overexpression assessed on histological sections of a retrospective series of 353 invasive breast carcinomas (294 invasive ductal carcinomas not otherwise specified, eight mucinous and 51 invasive lobular carcinomas), with a median follow-up of 10 years, using the polyclonal antibody CM1 and a cut-off score of > 10% of stained nuclei. p53 overexpression was not associated with invasive lobular or mucinous carcinomas (1.9% and 0% positive cases, respectively). In invasive ductal carcinomas (20.4% positive cases), there was a significant association between p53 overexpression and overall survival (P = 0.002), disease free interval (P = 0.005) and histological grade (P < 0.0001). Moreover, a statistically significant negative correlation between p53 immunoexpression and survival was found in high-grade (P = 0.016), pT1/2 (P = 0.001/0.018), and pN0 (P = 0.001) tumours, indicating its usefulness as a marker of prognosis in these subgroups of invasive ductal breast carcinoma.