Negative Progesterone Receptor Research Articles

Fascin expression predicts an aggressive clinical course in patients with advanced breast cancer

Fascin is an actin cross-linking protein, which regulates actin dynamics and filopodia or spike formation, as well as the epithelial-mesenchymal transition, and has been implicated in cell motility. Although, fascin is pivotal in mediating the aggressive behaviour of various types of cancer, its prognostic significance according to tumour stage has yet to be evaluated. Therefore, the present study investigated fascin expression in 194 patients diagnosed with invasive ductal carcinoma of the breast between 2000 and 2005. Fascin protein expression levels were evaluated by immunostaining on a tissue microarray, and the association between fascin expression and various clinicopathological parameters was analysed. Fascin expression was significantly correlated with various clinicopathological parameters, including high histological grade, tumour necrosis, resistance to adjuvant therapy, high expression of p53 and Ki-67 and specific therapeutic markers (oestrogen and progesterone receptor negativity; all P<0.05). Furthermore, univariate and multivariate analyses identified a significant association between fascin expression, and poor disease-free and overall survival, in late-stage breast cancer (all P<0.05). Therefore, fascin may be crucial in predicting aggressive tumour behaviour, particularly in patients with advanced-stage disease that has acquired the properties of migration and invasion.

Clinicopathologic Characteristics of Oestrogen Receptor-Positive/Progesterone Receptor-Negative/Her2-Negative Breast Cancer According to a Novel Definition of Negative Progesterone Receptor Status: A Large Population-Based Study from China.

PurposeA lack of progesterone receptor (PgR) expression in oestrogen receptor-positive (ER+) tumours is associated with worse survival. PgR status is usually defined as positive or negative using 1% positive nuclei as a cut-off point. In this study, we aimed to assess the clinicopathologic characteristics of ER+/PgR-/HER2- tumours by comparing them with ER+/PgR+/HER2- tumours using a PgR cut-off point of 20% as a divisive criterion.MethodsWe analysed 1,522 patients with primary breast cancer who had undergone surgery at the Cancer Center of Fudan University between 2012 and 2014. Age, grade, tumour size, lymph node status and lymphovascular invasion were assessed. Multinomial logistic regression, linear regression and chi-square test models were applied to assess associations between ER, PR and clinical features.ResultsER+/PgR-/HER2- tumours showed poorer clinicopathologic characteristics relative to ER+/PgR+/HER2- tumours using a PgR threshold of 20% instead of 1%. The clinicopathologic characteristics did not differ between tumours with purely negative PgR expression and tumours with a PgR percentage ranging from 1% to 19%. The prognostic significance of PR expression appeared more pronounced in patients under a high Ki-67 status than those under a low Ki-67 status.ConclusionsBased on these findings, we propose the use of a novel threshold of 20% to define PgR status. Nevertheless, the impact of this new criterion on patient management and clinical treatment requires additional study.

Abstract P2-03-05: The heterogeneous clinical behavior of luminal breast cancers is associated with different mutational landscapes

Abstract Background: Genomic studies in estrogen-receptor (ER) positive breast cancer showed few commonly mutated genes and suggested that tumors with higher background of mutation rate were enriched in intrinsic aromatase-inhibitor-resistant tumors (Ellis MJ Nature 2012). Different combinations of proliferation (PROL) and estrogen-receptor related genes (ERS) allow the prediction of the risk of recurrence and endocrine treatment sensitivity (Bianchini G Breast Cancer Res 2013). The highPROL/lowERS is enriched in intrinsic endocrine resistant tumors. We explored whether different genomic alterations could contribute to the different clinical behavior of groups defined by PROL and ERS. Method: We defined luminal breast cancers by PAM50 and retrieved gene expression and whole-exome sequencing data of 324 luminal breast cancer cases from the Cancer Genome Atlas (TCGA). PROL and ERS were calculated as previously reported. Low and high expression groups were defined by median cut-points (Bianchini G Breast Cancer Res 2013). As a metric of the background mutation rate of each sample, we used the overall mutation frequency in whole exome (Kandoth C Nature 2013). The most frequently mutated genes (TP53, PIK3CA, MAP2K4, MAP3K1, GATA3, CDH1) were also assessed for their association with PROL and ERS defined groups. Results: The average number of mutations (Nmut) was 44.4 (5-366). High and low proliferation groups had different mutation frequency (average Nmut 53.6 and 35.3, respectively; p=6.1E-06). LowERS group also had a higher Nmut (average Nmut 54.4 vs. 34.5 in high ERS; p=1.2E-06). Notably, neither high or low expression of the ER gene (p=0.89), nor positive and negative progesterone receptor by immunohistochemistry (p=0.69) were associated with Nmut. LowERS group was associated with higher Nmut in luminal A (p=0.005) and B (p=0.006) subtypes. The group with highPROL and lowERS had the highest Nmut (average 62.1), which was not significantly different from basal-like tumors. HighPROL/lowERS was significantly associated with higher frequency of mutTP53 (32.3%) and lower rates of mutPIK3CA (28.1%), mutMAP2K4 and/or mutMAP3K1 (7.2%) and mutCHD1 (3.1%). The lowPROL/highERS had the lowest Nmut (average 29.8), with the higher rate of mutPIK3CA (61.4%) and mutMAP2K4 and/or mutMAP3K1 (29.2%), and lower rate of mutTP53 (6.7%). Discussion: In ER-positive breast cancers, the highPROL/lowERS group, which is characterized by the highest risk of early recurrences despite endocrine treatment and chemotherapy, poor down-regulation of Ki67 after neoadjuvant aromatase-inhibitor (AI), but the highest rate of pCR after neoadjuvant chemotherapy, was also associated with the highest frequency of mutations, of the same order of magnitude of basal-like tumors. This data is in line with the observation that the number of mutations is linked to intrinsic resistance to neoadjuvant AIs (Ellis MJ Nature 2012). These data could be also informative for interpreting results of the combination of everolimus and endocrine therapy (exemestane or tamoxifen) that suggest higher efficacy in tumors with acquired endocrine resistance (lower mutation rate) compared to intrinsic or primary endocrine resistance (higher mutation rate). Citation Format: Giampaolo Bianchini, Eugenia Galeota, Tingting Jiang, Maurizio Callari, Milvia Zambetti, Takayuki Iwamoto, Thomas Karn, Christos Hatzis, Lajos Pusztai, Luca Gianni. The heterogeneous clinical behavior of luminal breast cancers is associated with different mutational landscapes [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-03-05.

Abstract P3-04-04: The prognostic effect of a negative progesterone receptor (PgR) by immunohistochemistry (IHC) in luminal HER-2 negative breast cancer (BC) by age at diagnosis: 10 years follow-up study

Abstract Background After menopause, an absent PgR predicts distant metastases (DM) in patients with ER positive HER-2 negative invasive BC. Yamamoto et al. (JCO 2013) questioned the prognostic effect of PR in premenopausal patients, as ovarian estrogens might affect PgR expression. We investigated the effect of PgR on DM free interval (DMFI) and BC specific survival (BCSS) by age at diagnosis as surrogate marker for menopause. Patients and methods Retrospective data of consecutive patients with a primary operable ER positive HER-2 negative BC (1/1/2000 - 31/12/2009) were retrieved from our prospectively managed database. Cases that received neo-adjuvant chemotherapy or were operated in another hospital, were excluded. BC with missing values for PgR, grade or lymph node status and/or lost to follow-up were excluded in some subgroup analyses. A multivariate (MV) competing risk model for DMFI and BCSS was established considering age at diagnosis (age ≤50 yrs vs &amp;gt;50 yrs), PgR status, tumor grade (1-3), tumor size (mm) and lymph node status (neg/pos). Steroid receptors were considered positive if ≥1% stained on IHC. Differential prognostic effects of these variables according to PR or age were tested by means of interaction effects. Only significant interactions were included. Results We included 3326 BCs (8 with missing PgR status); 2911 (87.5%) PgR positive [870 (26.2%) ≤50 yrs and 2041 (61.4%) &amp;gt;50 yrs] and 407 (12.5%) PgR negative [68 (2.0%) ≤50 yrs and 339 (10.2%) &amp;gt;50 yrs]. In absolute numbers and compared to PgR positive BCs, PgR negative cases &amp;gt;50yrs had more DM if grade 3 and more BCSS for all grades. In BC ≤50 yrs these differences were not found, but only 7.2% were PgR negative (Tab.1). Results from the MV models showed a significant interaction with PgR on DMFI and BCSS (respectively p=0.01 and p=0.002) resulting in higher risk of DM (HR, 1.9; 95% CI, 1.4-2.7; p=0.001) and BCSS (HR, 2.4; 95% CI, 1.6-3.7; p&amp;lt;0.001) when PgR was absent for BCP &amp;gt;50 yrs. This difference in DMFI and BCSS according to PgR status was not found in BC ≤50 yrs (Tab.2). Table 1. Number of BCP with distant metastasis and BCSS according to age (yrs), PgR status(+/-) and tumor grade.Distant Metastatic DiseaseBreast Cancer Specific Survival≤50 yrs&amp;gt;50 yrs≤50 yrs&amp;gt;50 yrsPgR+PgR-PgR+PgR-PgR+PgR-PgR+PgR-Grade 1-245/607 (7.4%)5/42 (11.9%)66/1512 (4.4%)11/226 (4.9%)28/607 (4.6%)2/42 (4.8%)33/1511 (2.1%)7/226 (3.1%)Grade 350/263 (19.0%)4/26 (15.4%)69/529 (13.0%)35/113 (31.0%)31/263 (11.8%)2/26 (7.7%)44/529 (8.3%)26/113 (23.0%) Table 2. Results of MV model including significant main effects and interaction effects between tumor characteristics, age and PgR status on DMFI and BCSS. 95% Confidence interval InteractionsHazard ratioLower limitUpper limitP-valueA. BMFIPgR Neg. vs Pos. |-50yrs0.7840.3951.5560.4869PgR Neg. vs Pos. |+50yrs1.9371.3862.7090.0001B. BCSSPgR Neg. vs Pos. |-50yrs0.5750.2081.5880.2854PgR Neg. vs Pos. |+50yrs2.4391.6203.672&amp;lt;.0001 Conclusions A negative PgR is only prognostic for DMFI and BCSS in women aged &amp;gt;50 at diagnosis. There was no difference of PgR for these endpoints in patients ≤50 yrs at diagnosis but PR negativity is rare in this age group. Citation Format: Siel JAR Olbrecht, Kathleen Van Asten, Annouschka Laenen, Chantal Remmerie, Wildiers Hans, Floris Guiseppe, Christiaens Marie-Rose, Vergote Ignace, Neven Patrick. The prognostic effect of a negative progesterone receptor (PgR) by immunohistochemistry (IHC) in luminal HER-2 negative breast cancer (BC) by age at diagnosis: 10 years follow-up study [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-04-04.

Abstract P6-08-05: Prognostic value of the progesterone receptor by proliferation rate in patients with luminal HER2 negative breast cancer

Abstract Background Estrogen receptor (ER) positive, HER2 negative breast cancer (BC) can be classified into luminal A and luminal B-like tumors according to tumor grade. Several evidences point to the fact that IHC expression of the progesterone receptor (PR) has prognostic value. In this study, we assess to what extent a negative PR in luminal A BCs increases the risk of distant metastasis and whether or not luminal B BC might have a good prognosis if PR is positive. Patients and methods Women with primary operable ER positive, HER2 negative BC treated at University Hospitals Leuven between 2000 and 2009 were retrieved from our database. So called luminal A tumors were defined as grade 1-2 BC, whereas so called luminal B BC were defined as grade 3 BC. Distant metastasis free interval (DMFI) and breast cancer specific survival (BCSS) were investigated by their PR status. PR was considered negative if the semi-quantitative Allred score was 0-2. Before 2003 the semi-quantitative H-score was used and a score &amp;lt;50 was considered negative. Statistical analysis for DMFI was performed using the cox proportional hazards regression. For BCSS we used Fine &amp; Gray’s competing risk regression. Covariates were age at diagnosis, tumor size, lymph node status, and therapy. Results In total, 3294 patients from Leuven were analyzed. From this cohort, 285 patients experienced metastases (8.7%) and 172 patients died of BC (5.2%). Details are shown in Table 1. The median age at diagnosis was 58 years with ages ranging from 22 to 95 years, 2358 patients (71.6%) were aged above 50 at diagnosis. The median follow-up period was 8.1 years. Table 1: Number of patients that metastasized and died of BC by luminal subgroup and PR status. MetastasesBC-related deathluminal APR positive110/2103 (5%)61/2103 (3%) PR negative16/267 (6%)9/267 (3%)luminal BPR positive120/786 (15%)75/786 (10%) PR negative39/138 (28%)27/138 (20%)BC: breast cancer, PR: progesterone receptor In Leuven, the reduction in risk of metastasis in patients with PR positive luminal A and luminal B BC was respectively 14% (Hazard ratio (HR): 0.86, 95% confidence interval (CI) 0.52-1.51) and 47% (HR: 0.53, 95% CI 0.37-0.78) compared with PR negative tumors. PR positive luminal A and luminal B BC patients had a 16% (HR: 0.84, 95 % CI 0.42-1.69) and 53% (HR: 0.47, 95% CI 0.30-0.75) reduction in the risk of BC-related death compared with PR negative tumors respectively. The same analysis was also carried out for postmenopausal patients (older than 50 years) only. In this subcohort, PR positivity was associated with a 9% (HR 0.91, 95% CI 0.50 to 1.83) and 59% (HR 0.41, 95% CI 0.28 to 0.63) reduction in the risk of metastatic events in luminal A and luminal B lesions, respectively. For BCSS, a 31% (HR 0.69, 95% CI 0.30 to 1.57) and 66% (HR 0.34, 95% CI 0.20 to 0.57) reduction in the risk of BC-related death for respectively PR positive luminal A and luminal B BC patients was found. Conclusion These results suggest that the prognostic effect of PR in primary operable BC depends on the tumor grade. Compared with luminal PR negative BC, PR positivity improves outcome more in luminal B than in luminal A lesions. Citation Format: Kathleen Van Asten, Ben Van Calster, Anneleen Lintermans, Olivier Brouckaert, Giuseppe Floris, Hans Wildiers, Patrick Neven. Prognostic value of the progesterone receptor by proliferation rate in patients with luminal HER2 negative breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P6-08-05.

Clinical pathological characteristics of breast cancer patients with secondary diabetes after systemic therapy: a retrospective multicenter study.

The objective of this study was to investigate the clinical pathological characteristics of breast cancer (BC) patients with secondary diabetes after systemic therapy without preexisting diabetes. A total of 1434 BC patients received systemic therapy and were analyzed retrospectively. Fasting plasma glucose (FPG) levels were monitored prior to the treatments, during the course of systemic therapy, and at the follow-up visits. Cox regression models were used to estimate the associations between the clinical pathological characteristics of BC and the cause-specific hazard of developing secondary diabetes. Among the 1434 BC patients, 151 had preexisting type 2 diabetes. Of the remaining 1283 patients with normal FPG levels prior to the systemic therapy, 59 developed secondary diabetes and 72 displayed secondary impaired fasting glucose (IFG) over a mean follow-up of 41 months. The prevalence of secondary type 2 diabetes in BC patients was 4.6 % (59/1283), which was obviously higher than that of the normal control group (1.4 %, P < 0.001). The percentage of older patients (P < 0.05), menopausal patients (P < 0.001), and obese patients (P < 0.01) tended to be lower in the secondary diabetic group. In addition, these patients with secondary diabetes had later pathological stages (P < 0.01), more lymph node metastasis (P < 0.05), negative estrogen receptor (ER) expression (P < 0.05), and smaller size of tumors (P < 0.05). After adjusting for age and BMI, the risk of developing secondary diabetes and IFG in subjects with later pathological stage BC (hazard ratio (HR) = 1.623; 95 % confidence interval (CI) 1.128–2.335 (P < 0.01)), negative progesterone receptor (PR) expression (HR = 0.530; 95 % CI 0.372–0.755 (P < 0.001)), positive human epidermal growth factor receptor 2 (HER2) expression (HR = 1.822; 95 % CI 1.230–2.700 (P < 0.01)), and more lymph node metastasis (HR = 1.595; 95 % CI 1.128–2.258 (P < 0.01)) was significantly higher. In conclusion, this study shows that an increase in the incidence of diabetes among breast cancer survivors after systemic therapy, especially the patients with later pathological stages, more lymph node metastasis, negative hormone receptor expression, and positive HER2 expression. Our study suggests that greater diabetes screening and prevention strategies among breast cancer patients after systemic treatment are needed in China.

Loss of heterozygosity in thyroid hormone receptor beta in invasive breast cancer.

Loss of heterozygosity (LOH) on chromosome arm 3p, where the gene of thyroid hormone receptor beta (THRB) is located, has been reported in breast cancer. Although some studies performed in vitro have suggested that THRB could act as a tumor suppressor in breast cancer development, there is still no unequivocal evidence to support this. To determine the role of LOH in breast tumor development, the LOH of THRB and its proximal microsatellite markers D3S1293, D3S3659, D3S3700, D3S2307 and D3S2336 was investigated in a genomic region spanning ~3.3 Mb in tumor specimens and in corresponding normal tissues of 74 invasive breast cancer patients. The association was analyzed between LOH in microsatellite markers and clinicopathological characteristics. LOH was detected in D3S1293 (36.7%), THRB (59.4%), D3S3659 (37.5%) and D3S3700 (55.2%) among the informative cases, while LOH was not detected in D3S2307 and D3S2336. Cases exhibited LOH of 52.8%-71.4% if any 2 markers were combined and analyzed out of the first 4 microsatellite markers. LOH in THRB was associated with negative estrogen receptor (ER), negative progesterone receptor (PR), both negative estrogen receptor and progesterone receptor (HR) and human epidermal growth factor receptor-2 (HER2) and lymph node metastasis (p = 0.0001, p = 0.005, p = 0.001 and p = 0.018). The association with negative PR in LOH in THRB and/or D3S1293 was pronounced (p&lt;0.0001). LOH in D3S3700 showed an association with lymph node metastasis (p = 0.014). This association was enhanced if D3S3700 was combined with THRB or D3S3659 (p = 0.0004, p = 0.0002). LOH in THRB and its proximal microsatellite markers may play a role in tumorigenesis and development in invasive breast cancer.

Lysophosphatidylcholine acyltransferase 1 (LPCAT1) upregulation in breast carcinoma contributes to tumor progression and predicts early tumor recurrence.

Breast cancer is the most common cancer in women worldwide. Aberrant lipid metabolism is an established hallmark of cancer cells. The recently isolated lysophosphatidylcholine acyltransferase 1 (LPCAT1), the most important enzyme in membrane biogenesis, has been currently implicated in cancer development and progression. The published literature lacks comprehensive reports on LPCAT1 expression in breast cancer and its impact on patients' outcome. We evaluated the immunohistochemical expression of LPCAT1 in 80 primary breast carcinomas, 24 metastatic lymph nodes, and 30 non-neoplastic breast tissue specimens and statistically analyzed the association between LPCAT1 expression and clinicopathological variables and patients' outcome. LPCAT1 protein was significantly upregulated in primary breast carcinoma and showed a significant ascending pattern being the lowest in normal breast tissues, relatively increased in fibrocystic disease, and the highest in primary carcinoma. LPCAT1 expression was significantly higher at tumor's advancing edge and correlated positively with tumor's grade and TNM stage. Compared to primary tumor, LPCAT1 expression was significantly lower in ductal carcinoma in situ and significantly higher in metastatic lymph nodes. LPCAT1 overexpression was significantly associated with increased proliferative activity, negative estrogen receptor (ER) and progesterone receptor (PR) status, positive human epidermal growth factor receptor 2 (HER2) status, as well as triple-negative and HER2 disease molecular subtypes. Multivariate analysis showed that advanced stage, high grade, and LPCAT1 overexpression were independent predictors of early tumor recurrence. We conclude that LPCAT1 is implicated in breast cancer pathogenesis, evolution, and progression and appears to play a potentially crucial role as a determinant of local invasiveness and metastasis. LPCAT1 is an independent predictor of early tumor recurrence of breast carcinoma and represents a novel prognostic biomarker that reflects underlying biological alterations and thus constitutes a potentially promising target for new therapeutic strategies.

Associations of polymorphisms in microRNAs with female breast cancer risk in Chinese population.

Polymorphisms in microRNA (miRNA) have been discussed to be associated with breast cancer risk; however, the conclusions were always inconsistent in different ethnicities. This case-control study enrolled 450 breast cancer cases, and 450 health controls was carried out to investigate the association between six polymorphisms in miRNAs and breast cancer risk. Sequenom MassARRAY was used to detect the polymorphisms in miRNAs, and the immunohistochemistry assay was applied to detect the expression of estrogen receptor (ER) and progesterone receptor (PR) in cancer tissue. The data showed that the 3746444 GG was associated with increased breast cancer risk (adjusted odds ratio (OR) = 1.71, 95 % confidence interval (CI) = 1.16-2.52) and that rs2292832 CC (adjusted OR = 0.54, 95 % CI = 0.34-0.85) was associated with decreased breast cancer risk. In addition, menopausal status subgroup analysis revealed that rs3746444 GG (adjusted OR = 2.34, 95 % CI = 1.31-4.15) and GA/GG (adjusted OR = 1.60, 95 % CI = 1.08-2.37) genotypes were associated with increased breast cancer risk for the subgroup of women with premenopausal status, respectively. Moreover, rs2910164 GG (adjusted OR = 1.84, 95 % CI = 1.07-3.15) and CG/GG (adjusted OR = 1.47, 95 % CI = 1.01-2.15) genotypes were associated with increased breast cancer risk in the postmenopausal status subcohort, respectively. Furthermore, rs3746444 AG (adjusted OR = 1.61, 95 % CI = 1.06-2.45) and AG/GG (adjusted OR = 1.49, 95 % CI = 1.02-2.18) genotypes were observed to be associated with increased risk of lymph node involvement and breast cancer with negative PR expression, separately. In short, rs3746444 was associated with breast cancer risk, especially for women with premenopausal status, and rs2910164 CG and CG/GG genotypes were associated with breast cancer risk for the women with premenopausal status.

Aberrant methylation of imprinted genes is associated with negative hormone receptor status in invasive breast cancer

Epigenetic regulation of imprinted genes enables monoallelic expression according to parental origin, and its disruption is implicated in many cancers and developmental disorders. The expression of hormone receptors is significant in breast cancer because they are indicators of cancer cell growth rate and determine response to endocrine therapies. We investigated the frequency of aberrant events and variation in DNA methylation at nine imprinted sites in invasive breast cancer and examined the association with estrogen and progesterone receptor status. Breast tissue and blood from patients with invasive breast cancer (n = 38) and benign breast disease (n = 30) were compared with those from healthy individuals (n = 36), matched with the cancer patients by age at diagnosis, ethnicity, body mass index, menopausal status and familial history of cancer. DNA methylation and allele-specific expression were analyzed by pyrosequencing. Tumor-specific methylation changes at IGF2 DMR2 were observed in 59% of cancer patients, IGF2 DMR0 in 38%, DIRAS3 DMR in 36%, GRB10 ICR in 23%, PEG3 DMR in 21%, MEST ICR in 19%, H19 ICR in 18%, KvDMR in 8% and SNRPN/SNURF ICR in 4%. Variation in methylation was significantly greater in breast tissue from cancer patients compared with that in healthy individuals and benign breast disease. Aberrant methylation of three or more sites was significantly associated with negative estrogen-alpha (Fisher's exact test, p = 0.02) and progesterone-A (p = 0.02) receptor status. Aberrant events and increased variation in imprinted gene DNA methylation, therefore, seem to be frequent in invasive breast cancer and are associated with negative estrogen and progesterone receptor status, without loss of monoallelic expression.

Prognostic Impact and Clinicopathological Correlation of CD133 and ALDH1 Expression in Invasive Breast Cancer.

PurposeCD133 and aldehyde dehydrogenase 1 (ALDH1) expression are reliable poor-prognosis markers associated with the presence of adverse biomarkers and subtypes of breast cancer. The aim of our study was to investigate and compare the clinical impact of CD133 and ALDH1 expression in invasive breast cancer.MethodsA total of 291 consecutive patients with invasive breast cancer who underwent breast cancer operations from 2005 to 2010 at a single institution were included in this retrospective review. CD133 and ALDH1 expression were determined by immunohistochemistry.ResultsCD133 and ALDH1 expression were positive in 24.7% and 22.0% of the patients, respectively, and were associated with tumor size, cancer stage, estrogen receptor negativity, nonluminal subtype, triple-negative breast cancer, and recurrence. CD133 expression was significantly associated with lymph node metastasis, progesterone receptor negativity, human epidermal growth factor receptor 2 positivity, chemotherapy, and poor disease-free (p=0.002) and overall survival (p=0.014), but ALDH1 expression was not. Cancer stage (p<0.001) was an independent prognostic factor for disease-free survival in multivariate analysis. Cancer stage (p<0.001) and receipt of radiotherapy (p=0.045) were independent prognostic factors for overall survival in multivariate analysis.ConclusionCD133 or the combination of CD133 and ALDH1 expression were more widely associated with the presence of adverse biomarkers and subtypes of breast cancer, compared to ALDH1 expression alone, and these markers may have a potential predictive role and be a helpful tool in the management for patients with invasive breast cancer.

Correlation of carbonic anhydrase IX expression with clinico-morphological parameters, hormonal receptor status and HER-2 expression in breast cancer.

The hypoxia-inducible protein carbonic anhydrase IX is widely expressed in most human cancers, including breast carcinomas. CA IX attracts significant interest due to its strong association with neoplasms and its absence from corresponding normal tissues, suggesting its potential to serve as apromising diagnostic biomarker. This protein comes into the limelight also as avaluable prognostic and predictive parameter. Immunohistochemically, we examined the expression of this protein in 84 cases of invasive breast carcinoma to determinate the association with clinico-morphological and biological parameters such as age of patients, grade, stage and size of primary tumor, lymph node metastasis, vascular invasion as well as hormone receptor status and HER-2 expression. In each case, the subcellular localization of CA IX antigen, the intensity of staining and the percentage of labeled cells were assessed. Overall, CA IX was expressed in 34 cases (40.5%). The statistical analysis revealed asignificant correlation between subcellular localization of CA IX and the age of patients. Furthermore, significant correlations were also found between the grade, estrogen and progesterone status and all immunohistochemical characteristics of CA IX expression (the subcellular localization of CA IX antigen, the intensity of staining and the percentage of labeled cells). We point out that mostly membrane or combined membrane and cytoplasmic positivity together with ahigher intensity of CA IX immunoreactivity are associated with poor prognostic parameters, such as tumor grade 3 and also with negative estrogen and progesterone receptor status which may influence therapeutic approach. However, no significant correlations were shown with remaining clinico-morphological and biological parameters. We next investigate the relationship between CA IX expression in the group of invasive ductal carcinomas and the group of invasive lobular carcinomas and other less frequent types of breast carcinoma. There was, however, no significant difference. Our results suggest that moderate to strong membrane and combined membrane and cytoplasmic localization of CA IX may represent avaluable tumor biomarker as well as apromising prognostic and predictive parameter in invasive breast cancer. breast carcinoma, carbonic anhydrase IX, immunohistochemistry, clinico-morphological and biological parameters.

Association of clinical, histopathological and immunohistochemical prognostic factors of invasive breast tumors and thermographic findings

BackgroundThe purpose of the paper was to analyse and compare infrared thermographic findings with clinical, histopathological and immunohistochemical prognostic parameters in female patients with invasive breast tumor. Methods: A pre-operative thermography was made in 75 female patients with breast invasive tumors. The study analysed an individual impact of each clinical, histopathological (HP), and immunohistochemical (IHC) factors on thermographic findings, the joint impact of all factors and combined impacts according to the IHC phenotypes. Results: Statistically significant difference of thermographic findings between healthy and affected breast was found for positive human epidermal growth factor receptor-2 (HER-2) tumors, negative progesterone receptors (PR−) and a high proliferative activity (Ki-67>30%) (p<0.05). Dependent on the IHC phenotype, temperatures varied from the coldest (ER+, PR+, HER-2−) tumor towards the warmest tumors (ER+, PR−, HER-2+).Conclusions: According to the results of this study, the increased temperature was more dependent on the IHCphenotypes of the tumor than on other clinical and histopathological prognostic factors. Moreover, tumor size had no influence on increased temperature.

Protein Expression and Codon 72 Polymorphism of TP53 Gene in Triple Negative Breast Cancer

A subgroup of tumor that has received attention is triple-negative breast cancer (TNBC), which presents phenotype of negative estrogen receptor, negative progesterone receptor and has no overexpression of HER2. TP53 acts as a tumor suppressor limiting the proliferation of damaged cells. A polymorphic site (rs1042522) of TP53 encodes either an arginine or a proline amino acid, but its biological significance remains unclear. This study aimed to investigate this variant and its expression in search for a possible involvement in TNBC susceptibility and clinical outcome. Genetic polymorphism was evaluated in 50 patients and 115 controls by PCR based methodology and immunohistochemistry was done with monoclonal antibody. Case-control study showed no positive or negative association (OR= 0.95; CI95%= 0.48-1.89). Comparison of genotypes and clinical outcome showed no significant results. Despite most of patients presented p53 positive staining by immunohistochemistry, there was no significant association in relation to prognostic parameters. Results demonstrated a lack of association between codon 72 polymorphism, susceptibility and prognosis of TNBC. Immunohistochemistry analysis should be done more carefully, since most of the patients had the somatic mutation of p53, which could be an indicator of prognostic value in TNBC.

Abstract B32: Heterogeneity of breast cancer clinical characteristics and outcomes in U.S. black women - effect of place of birth

Abstract Background: Breast cancer mortality in black women is disproportionately high; reasons for this phenomenon are still unclear. In addition to socioeconomic factors, the biology of the tumor may play a role. Methods: We analyzed 1,097 incident invasive breast cancer cases diagnosed between 2000 and 2010 in black US women from Long Island and Brooklyn. Kaplan-Meier curves were calculated and fitted for cancer mortality; when data were stratified, the log-rank test was used to assess statistical differences in survival across strata. A multivariable Cox hazard model was applied to assess the independent contributions to survival of several factors such as age, histology, stage at diagnosis, and hormone receptor status. Results: Thirty-five percent of women had an estrogen receptor (ER) negative tumor, 46% a progesterone receptor (PR) negative tumor. ER, PR negative tumors were diagnosed at an earlier age (55.8 vs 55.3 yrs), at a later stage (p= 0.06), were larger in size (p=0.04), and more frequently treated with neoadjuvant chemotherapy (p=0.06) than ER, PR positive tumors. Determinants of shorter survival were: ER, PR negativity (HR: 2.2, 95% CI: 1.4-3.4), age, and stage at diagnosis (HR: 2.0; 95% CI: 1.5-2.7). ER, PR negative women born outside of the US experienced significantly worse survival than ER, PR negative women who were born in the US. There were no differences in survival based on therapeutic strategy (radiation, chemotherapy, or surgery). Conclusions: ER, PR negative tumors in black women born outside the US, mainly in the Caribbean, are biologically more aggressive than the same tumor size and age matched tumors in black women born in the US. Our study suggests that environmental exposures in the country of origin may impact on host cancer interactions and cancer outcome. Citation Format: Marlene Camacho-Rivera, Tricia Kalwar, Jasotha Sanmugarajah, Iuliana Shapira, Emanuela Taioli. Heterogeneity of breast cancer clinical characteristics and outcomes in U.S. black women - effect of place of birth. [abstract]. In: Proceedings of the Sixth AACR Conference: The Science of Cancer Health Disparities; Dec 6–9, 2013; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2014;23(11 Suppl):Abstract nr B32. doi:10.1158/1538-7755.DISP13-B32

Correlation of Primary Tumor FDG Uptake with Clinicopathologic Prognostic Factors in Invasive Ductal Carcinoma of the Breast.

The purpose of this study was to investigate the correlation of primary tumor FDG uptake to clinicopathological prognostic factors in invasive ductal carcinoma of the breast. We retrospectively reviewed 136 of 215 female patients with pathologically proven invasive ductal breast cancer from January 2008 to December 2011 who underwent F-18 FDG PET/CT for initial staging and follow-up after curative treatment with analysis of estrogen receptor (ER), progesterone receptor (PR) and human epithelial growth factor receptor 2 (HER2). The maximum standardized uptake value (SUVmax) of the primary breast tumor was measured and compared with hormonal receptor and HER2 overexpression status. The high SUVmax of primary breast tumors is significantly correlated with the clinicopathological factors: tumor size, histologic grade, TNM stage, negativity of ER, negativity of PR, HER2 overexpression and triple negativity. The recurrent group with non-triple negative cancer had a higher SUVmax compared with the non-recurrent group, though no significant difference in FDG uptake was noted between the recurrence and non-recurrent groups in subjects with triple-negative cancer. Lymph node involvement was the independent risk factor for cancer recurrence in the multivariate analysis. In conclusion, high FDG uptake in primary breast tumors is significantly correlated with clinicopathological factors, such as tumor size, histologic grade, TNM stage, negativity of the hormonal receptor, HER2 overexpression and triple negativity. Therefore, FDG PET/CT is a helpful prognostic tool to direct the further management of patients with breast cancer.

Methylation profiling of ductal carcinoma in situ and its relationship to histopathological features.

IntroductionDNA methylation is a well-studied biomarker in invasive breast cancer, but its role in ductal carcinoma in situ (DCIS) is less well characterized. The aims of this study are to assess the methylation profile in DCIS for a panel of well-characterized genes that are frequently methylated in breast cancer, to investigate the relationship of methylation with pathological features, and to perform a proof-of-principle study to evaluate the practicality of methylation as a biomarker in diagnostic DCIS material.MethodsPromoter CpG island methylation for a panel of 11 breast cancer-related genes was performed by methylation-sensitive high resolution melting (MS-HRM). Formalin-fixed, paraffin-embedded (FFPE) biopsies from 72 samples of pure DCIS (DCIS occurring in the absence of synchronous invasive carcinoma), 10 samples of mixed DCIS (DCIS adjacent to invasive carcinoma), and 18 samples of normal breast epithelium adjacent to a DCIS lesion were micro-dissected prior to DNA extraction.ResultsMethylation was seen for all the tested genes except BRCA1. RASSF1A was the most frequently methylated gene (90% of DCIS samples) and its methylation was associated with comedo necrosis (p = 0.018). Cluster analysis based on the methylation profile revealed four groups, the highly methylated cluster being significantly associated with high nuclear grade, HER2 amplification, negative estrogen receptor (ER) α status, and negative progesterone receptor (PgR) status, (p = 0.038, p = 0.018, p <0.001, p = 0.001, respectively). Methylation of APC (p = 0.017), CDH13 (p = 0.017), and RARβ (p <0.001) was associated with negative ERα status. Methylation of CDH13 (p <0.001), and RARβ (p = 0.001) was associated with negative PgR status. Methylation of APC (p = 0.013) and CDH13 (p = 0.026) was associated with high nuclear grade. Methylation of CDH13 (p = 0.009), and RARβ (p = 0.042) was associated with HER2-amplification.ConclusionsDNA methylation can be assessed in FFPE-derived samples using suitable methodologies. Methylation of a panel of genes that are known to be methylated in invasive breast cancer was able to classify DCIS into distinct groups and was differentially associated with phenotypic features in DCIS.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-014-0423-9) contains supplementary material, which is available to authorized users.

PP105-SUN: Body Mass Index and Breast Cancer Subtype in Pre-Postmenopausal Women

Rationale: Obesity has a complex relationship to breast cancer risk that differs in premenopausal and postmenopausal women. Methods: This study aims to determine the association between Body Mass Index (BMI) and the distribution of breast cancer subtypes defined by Estrogen Receptor (ER), Progesterone Receptor (PR) and in preand postmenopausal breast cancers. A total of 105 female breast cancer patients were included in the analyses. The histological type of the tumor, ER and PR expression, age, body weight, height and menopausal status at diagnosis were investigated retrospectively. The patients with normal body weight had BMI 24.9 kg/m2, overweight had BMI between 25.0 and 29.9 kg/m2, and obese patients had BMI 30.0 kg/m2. Results: Median BMI was 27.098 kg/m2 (11.72 35.55) in the postmenopausal and 25.781 kg/m2 (14.84 40.60) in the premenopausal group (P< 0.065). BMI at diagnosis did not differ significantly between the menopausal age groups (P= 1.636). BMI was similar between the menopausal age both in preand postmenopausal and subtype breast cancer (P= 1.556 and P= 0.831, respectively). Premenopausal women with a BMI of 25.0 kg/m2 showed a propensity towards negative Estrogen Receptor and Progesterone Receptor (ER /PR ) tumors when compared to premenopausal women with a BMI of <25.0 kg/m2 (P= 0.648). Conclusion: The risk of breast cancer may be not associated with BMI in preand postmenopausal breast cancer patients in our study. However, obesity might be related to an increased risk of premenopausal hormone receptor-negative breast cancer. Further studies, with large figures, are needed for explanation of the credible mechanisms in the pathogenesis of premenopausal hormone receptor-negative breast cancer.

T3 levels in relation to prognostic factors in breast cancer: a population-based prospective cohort study.

BackgroundThe issue of a potential association between thyroid conditions/hormones and breast cancer has been studied extensively during the last decades but the results have been inconclusive and almost no studies have investigated breast cancer aggressiveness. We have previously found a positive association between prospectively measured levels of triiodothyronine (T3) and breast cancer incidence as well as breast cancer mortality. We now investigated prediagnostic T3 levels in relation to specific prognostic factors in breast cancer.MethodsThe Malmö Preventive Project is a population-based prospective cohort including 2185 women in whom T3 levels were measured at baseline. That is, total T3 levels were measured before a potential diagnosis of breast cancer. Mean follow-up was 23.3 years and 149 women in the study population were diagnosed with invasive breast cancer. Tumours were classified according to selected prognostic factors of breast cancer; i.e. grade, tumour size, lymph node metastasis, and hormonal receptor status. T3 was handled both as tertiles and as a continuous variable. A Cox’s proportional hazards analysis yielded hazard ratios with 95% confidence intervals. All analyses were also restricted to postmenopausal women.ResultsOverall there was a statistically significant association between T3 and “all” breast cancers. The adjusted Hazard Ratio (HR) in the third tertile, as compared to the first, was (1.61:1.07-2.43). There was a statistically significant positive association between the third T3 tertile and large tumours, i.e. > 20 mm, (3.17:1.20-8.36) and the occurrence of lymph node metastases, (4.53:1.60-12.83). Other prognostic factors positively associated with T3 were negative oestrogen receptor (ER) status, (3.52:1.32-9.41) and negative progesterone receptor (PGR) status, (3.52:1.42-8.75). The analyses of T3 as a continuous variable and analysis restricted to postmenopausal women, confirmed the results but also showed an association with smaller tumours and in postmenopausal women a contemporary association with negative lymph nodes.ConclusionsThis prospective study of serum T3 levels in relation to breast cancer aggressiveness is the first of its kind. We found statistically significant positive associations between higher prediagnostic T3 levels and larger tumours, occurrence of lymph node metastases, and negative ER and PGR status.

The expression and significance of five types of miRNAs in breast cancer.

BackgroundThis study aimed to investigate the expression and significance of 5 types of miRNAs in breast cancer to provide a theoretical and practical foundation for using these miRNAs in the diagnosis and treatment of breast cancer, thereby improving medical services.Material/MethodsStem-loop real-time RT-PCR was used to detect the expression levels of miR-145, miR-21, miR-10b, miR-125a, and miR-206 in 35 cases of breast cancer and adjacent normal breast tissues, and to analyze the relationship of miRNAs expression with clinicopathological features of breast cancer. The expression levels of estrogen receptor (ER) and progesterone receptor (PR) were examined by immunohistochemistry. Fluorescence in situ hybridization was used for the detection of HER-2 and TOP 2A.ResultsThe expression levels of miR-145, miR-125a, and miR-206 in breast cancer were lower than those in adjacent normal tissues. MiR-145 was negatively correlated with tumor size, lymph node metastasis, ER, HER-2, and TOP 2A (P<0.05), regardless of age, menstruation, and PR. MiR-125a was correlated with negative node status, negative HER-2 status (P<0.05), whereas tumor size, age, menstruation, ER, and PR were independent factors. MiR-206 expression was correlated with negative ER status, negative PR status, and negative HER-2 status (P<0.05), regardless of age, menstruation, lymph node metastasis, and TOP 2A. MiR-21 and miR-10b expression in breast cancer tissues was significantly higher than that in adjacent tissues (P<0.05). MiR-21 in post-menstrual patients with lymph node metastasis was highly expressed (P<0.05), and had no correlations with tumor size, ER, PR, and TOP 2A expression. MiR-10b expression was positively correlated with breast cancer tumor size, lymph node metastasis, and TOP 2A status (P<0.05), but had no correlations with age, menstruation, ER, PR, and HER-2.ConclusionsMiR-145, miR-21, miR-10b, miR-125a, and miR-206 may play important roles in breast cancer development and invasion.