bluuebunny/crossref_metadata_2025 huggingface dataset:
P1-07-15: The Reliability of Ki-67 Expression Assessment Using Core Needle Biopsy and Surgical Specimens of Invasive Breast Cancer: Can Ki-67 Change Predict Benefits of Preoperative Endocrine Therapy?
Authors: Mizuno Y , Takayanagi H , Sato K
Published: June 2012

Background: The use of Ki-67 change as a predictive marker in breast cancer has been widely investigated in patients treated with preoperative endocrine therapy. It has been assumed that Ki-67 expression determined using core needle biopsy and surgical specimens of invasive breast cancer is concordant. Many studies have suggested the concordance of the results obtained using core needle biopsy and surgical specimens for expression of the estrogen receptor (ER), progesterone receptor (PgR), and human epidermal growth factor receptor type2 (HER2) status in early-stage breast cancer. However, there was few study to compare Ki-67 expression in the core needle biopsy and surgical specimens. We conducted a retrospective study of Ki-67 expression assessment between the two types of specimens to answer this question.

Methods: A total of 269 patients underwent primary operations for early-stage breast cancer at Tokyo-West Tokushukai Hospital from August 2008 to May 2011. Among these patients, 169 patients who received core needle biopsy prior to operation were enrolled.

Patients with ductal carcinoma in situ and those with neoadjuvant chemotherapy were excluded. A cutoff value of 20% was used for Ki-67-positive criteria. We also examined ER, PgR, and HER2 expression and compared it with Ki-67 expression. Statistical significance for concordance rates between the two types of specimens was evaluated by Wilcoxon t-test. To evaluate the consequence of formalin and genetic heterogeneity, parameters such as operative method and tumor size were analyzed by χ2 analysis.

Results: The concordance rate between the two types of specimens for Ki-67 expression was 76%, and this was significantly lower than that for ER expression, which was 96%. The concordance rates for PgR and HER2 expression were 88% and 91%, respectively, and they were not significantly different from the rate for Ki-67 expression. 45 patients(34.6%) had received mastectomy in the Ki-67 concordant group, and 14 patients(34.1%) in the discordant group. 55 patients(42.0%) showed T2∼ tumor size in the concordant group, 13 patients(31.7%) in the discordant group. No significant differences of parameters such as operative method (mastectomy vs breast conserving surgery) and tumor size (T1 vs T2∼) were observed between the two patient groups.

Conclusion: The reliability of Ki-67 concordance rate assessment using core needle biopsy specimens was significantly lower than that of ER expression. A standard pathological assessment of Ki-67 expression might be needed for the possible use of Ki-67 as a predictive marker for preoperative endocrine therapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-07-15.
P1-08-05: Age and Survival in Women with Early Stage Breast Cancer: An Analysis Controlling for Tumor Subtype.
Authors: Partridge AH , Hughes ME , Ottesen R , Wong Y-N , Edge SB , Theriault RL , Blayney DW , Niland JC , Winer EP , Weeks JC , Tamimi RM
Published: June 2012

Background: Previous research has suggested that young age at diagnosis is an independent risk factor for breast cancer recurrence and death in women with early stage breast cancer. However, young women are more likely to have aggressive subtypes of breast cancer. No prior studies have adequately controlled for tumor phenotype, including HER-2/neu (HER2) status, in particular. Recent evidence has suggested that the prognostic effect of young age varies by tumor subtype.

Methods: We examined data from women with newly diagnosed Stage 1–3 breast cancer presenting to one of 8 NCCN centers between January 2000 and December 2007. Multivariate Cox proportional hazards models were used to assess the relationship between age and breast cancer specific survival, controlling for known prognostic factors and treatment. In addition, we conducted stratified analyses by estrogen receptor (ER) and HER2 status.

Results: 19,633 women with Stage 1–3 breast cancer eligible for analysis including 2,177 (11%) who were age 40 years or younger at diagnosis. Younger women were more likely to be non-white or Hispanic, more educated, employed, and to have higher stage, high grade, ER-negative, progesterone receptor (PR) negative, and HER2−positive disease, and treated with chemotherapy and trastuzumab (all variables P< 0.0001 by Chi-Square test). 5-year survival among younger women was 94.1 (95% Confidence Interval [CI] 92.9−95.3) and 96.3 (95% CI 95.9−96.6) for older women. In a multivariate Cox proportional hazards model controlling for sociodemographic, disease, and treatment characteristics, women age < 40 or younger at diagnosis had increased mortality compared to older women (Hazard Ratio [HR] 1.26, 95% CI 1.02−1.56). In stratified analyses, age 40 or less was associated with increased mortality among women with ER-positive disease (HR 1.44, 95% CI 1.01−2.05), but was not among those with ER-negative disease (HR 1.15, 95% CI 0.85−1.55). Younger age was associated with a statistically significant increase in mortality among women with HER2−negative disease (HR 1.29, 95% CI 1.00−1.68), but this difference did not reach statistical significance among those with HER2−positive disease (HR 1.30, 95% CI 0.82−2.09). Conclusions: The effect of age on short-term survival of women with early breast cancer appears to vary by breast cancer subtype, particularly ER status. Further research to elucidate differences in breast cancer biology and efficacy of therapy within tumor types by age is warranted.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-05.
P1-08-11: Differences in Recurrence Dynamics between Lobular and Ductal Invasive Breast Cancer.
Authors: Siesling S , Kwast ABG , Grandjean I , Ho V , van der Sangen MJC , Menke-Pluymers MBE , Tjan-Heijnen VCG
Published: June 2012

Introduction: Many studies investigated disease free survival after breast cancer treatment. A few studies focused on patterns of recurrence risk over time. However, they ignored the impact of histology. This study aimed to determine differences in recurrence dynamics on population-based data between patients with a ductal breast cancer (DBC) and lobular breast cancer (LBC).

Methods: All surgically treated women diagnosed in 2003–2004 with invasive DBC or LBC, with no distant metastases or second primary breast cancer were selected from the Netherlands Cancer Registry. The recurrence patterns were studied using the life-table method to estimate the hazard rate for the first recurrence with the worst prognosis (locoregional (LRR) or distant metastases), that is, the conditional probability of manifesting recurrence in a time interval, given that the patient is clinically free of any recurrence at the beginning of the interval. The wilcoxon test was used to determine differences in recurrence patterns between DBC and LBC.

Results: Of 16,231 women identified, 87% had DBC and 13% LBC. LRR was found in 4% of the DBC and in 3% of the LBC patients, whereas metastases were found in 11% and 10% of patients, respectively. Notably, in DBC patients a peak in recurrence rate was seen between 1–2 years after diagnosis, both for LRR and distant metastases. On the other hand, in LBC patients the recurrence pattern did not show major peaks: after 2 years the curve revealed an almost steady level for LRR and distant metastases. This recurrence pattern was significantly different (P=<0.001). Moreover, the influence of age, stage and treatment on recurrence differed between DBC and LBC. We also noticed a difference in localization of metastasis.

Conclusion: Our study showed an overall equal number of recurrences after DBC and LBC, but with a totally different recurrence pattern, which could have implications on follow-up of the patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-11.
P1-08-16: Benign Breast Disease (BBD) and Breast Cancer in African American Women.
Authors: Fehmi RA , Cote M , Ruterbusch J , Alosh B , Bandyopadhyay S , Albashiti B , Frost MH , Hartmann LC , Visscher DW
Published: June 2012

African American (AA) women have higher mortality rates from breast cancer (BrCa) and are diagnosed at younger ages than their Caucasian counterparts. Women who have had a benign breast biopsy are at increased risk of the disease, although less is known about the risk of BrCa associated with benign breast disease in AA women. We examined 1428 breast biopsies from AA women which occurred from 1997–2000 and assessed various pathologic markers including: apocrine metaplasia, ductal hyperplasia including atypia, evidence of cysts, duct ectasia, fibrosis, intra-ductal papilloma, sclerosing adenosis, columnar alteration, and involution (atrophy). These women were followed for later BrCa through the Metropolitan Detroit Cancer Surveillance System, part of the Surveillance, Epidemiology and End Results (SEER) program through 2008. Women who developed BrCa were compared to those in the cohort who did not, and to other AA women with BrCa in the SEER registry. AA women in our study were also compared to Caucasian women in the Mayo Clinic cohort. Differences in variables were assessed by chi-squared tests and 95% confidence intervals. Of the 1428 biopsies, 52 (3.6%) subsequent incident breast cancers were reported in SEER. The mean age at diagnosis was 59, and the mean time from biopsy to BrCa diagnosis was 6.1 years. Individuals with atypical ductal hyperplasia at biopsy (n=37, 2.6%) were more likely to develop breast cancer (n=7, 13.5%, p<0.01). No other pathologic variables were associated with increased risk. Women in our cohort with breast cancer did not differ from AA in the SEER database with respect to age at diagnosis, stage at diagnosis, or receptor positivity. Compared to the Caucasian women in the Mayo Clinic BBD cohort, AA women in our study were younger at biopsy (p<0.01), but had similar percentages of involution and atypia (p=0.50 and 0.15, respectively). Our preliminary findings among a relatively small group of AA women with prior benign breast biopsies and incident breast cancers suggest that results from the Mayo Cohort study are likely to apply to AA populations.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-16.
P1-08-20: Parity Interferes with the Effect of Age at Diagnosis on the Frequency Breast Cancers Are Triple-Negative.
Authors: Vannevel V , Brouckaert O , Leunen K , Amant F , Berteloot P , Dieudonne A-S , Laenen A , Wildiers H , Paridaens R , Van Limbergen E , Weltens C , Moerman P , Smeets A , Lambrechts D , Christiaens M-R , Vergote I , Neven P
Published: June 2012

Background Epidemiologic studies show an age related decrease in the frequency that breast cancers are triple negative (TN) (estrogen receptor, progesterone receptor and HER-2 negative). Parity increases the risk of TN breast cancer as a previous full-term pregnancy mainly protects against ER+ breast cancer. It is unknown whether this protective affect appears at all ages of breast cancer diagnosis. We study the frequency of triple negative breast cancers by parity and age at breast cancer diagnosis.

Methods We performed a retrospective case-case analysis including 1583 consecutive female patients with primary diagnosis of invasive breast cancer, < 51 years at diagnosis. We compared the frequency of TN tumors between parous (N = 1271) and nulliparous (N = 312) women in three age categories (21 to 30 yrs, 31 to 40 yrs and 41 to 50 yrs). For statistical analysis we used a logistic regression model.

Results We confirmed a decrease of TN breast cancer with age (p < 0.0001). This decrease is stronger pronounced in the nulliparous group. Parity favors TN tumors only in women > 40 years at breast cancer diagnosis.

Discussion: Although several other factors may affect the frequency breast cancers are TN, age at breast cancer diagnosis interacts with the effect of a previous pregnancy on the frequency breast cancers are TN. Breast cancers in young women (< age 40) may have been initiated long before pregnancy can induce a protective effect against ER-positive cases. The international BCAC has recently accepted to validate our UZ Leuven findings.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-20.
P1-08-24: Clinical and Pathologic Characteristics of Diabetic Breast Cancer Patients in a Tertiary Care Safety Net Hospital.
Authors: Loch MM , Eapen A , Ross AA , Rosenberg C , Blanchard RA
Published: June 2012

Background: Recent studies have indicated an increased risk of breast cancer in women with diabetes, and an increase in disease specific mortality in patients with diabetes and breast cancer. Clinical and pathologic features of diabetic women with breast cancer have not been described. We examine tumor characteristics in diabetic women in our ethnically diverse tertiary care hospital that may help explain the increased mortality seen in diabetic women diagnosed with breast cancer.

Methods: We expanded a pre-existing database to include all women diagnosed with breast cancer from 1998–2011. From the electronic medical record we documented age, body mass index (BMI), place of birth, self-identified race/ethnicity, date of diagnosis, T, N, M stage, grade, ER, PR, HER2 expression, presence of diabetes (DM) or thyroid disease at diagnosis, recurrence and treatment data including neoadjuvant, surgical, adjuvant and radiation therapy and adequacy of treatment. We determined associations using contingency tables and multivariate logistic regression.

Results: 480 cases were identified. Patients were racially diverse: 35% white [W], 41% black [B], 14% Hispanic and 10% other. 23% had DM, 47% were obese (body mass index [BMI] >30). Age in DM vs. non-DM were 63.327 vs. 57.729 (p= < 0.0001) and BMI in DM vs. non-DM were 32.749 vs. 29.433 (p<0.001). When exploring odds ratios (OR) of different tumor subtypes, none were statistically significant when controlling for age and BMI. The OR calculated were as follows: Triple negative 1.00 (confidence interval [CI]: 0.6−1.666, p = 0.9996), ER+ 1.063 (CI: 0.68−1.662, p=0.7877), PR+ 1.005 (CI: 0.67−1.506, p=0.9811), HER2+ 1.011 (CI: 0.578−1.768, p= 0.8535), recurrence: 1.090 (CI: 0.634−1.873, p= 0.7558). There were no significant interactions with DM and T, N stage when controlling for age and BMI. T stage: p=0.6962, N stage: p=0.7349 and recurrence p=0.5469 on recurrence controlling for age, BMI and grade: p=0.5462. There was no significant association between grade in DM vs. no DM with rates of grade in DM as follows: grade 1: 10.81%, grade 2: 49.55%, grade 3: 39.64% and in non-DM grade 1: 13.55%, grade 2: 45.8% and grade 3: 40.65%. Mean score difference p=0.7588. Conclusions: We did not find significant differences in our DM women compared with non-DM women when evaluating tumor subtype, T stage, N stage, grade or recurrence. In our patient population these factors do not play a role in the increased cause specific mortality in DM patients with breast cancer. Other factors must play a role in this and should be explored further.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-24.
P1-09-05: A 3′UTR Functional Variant in BRCA1: A Predictor of Poor Outcome in Breast Cancer.
Authors: Dorairaj JJ , Miller N , Newell J , Kerin MJ , Weidhaas JB
Published: June 2012

Background: MicroRNAs (miRNAs) are class of gene regulators which exert their effects through binding with partial complementarity to sequences in the 3'untranslated region (3'UTR) of the target mRNA. Single nucleotide polymorphisms (SNPs) in the 3'UTR of target mRNA have the potential to disrupt or create new illegitimate miRNA targets and have been associated with cancer predisposition as well as tumor biology. An association between rs8176318 and risk for familial breast and ovarian cancer as well as risk for triple negative breast cancer in African American women has previously been established. We aimed to evaluate the phenotypic effect of rs8176318 in a west of Ireland breast cancer cohort, and also its role as a biomarker of prognosis.

Methods: DNA from 727 unselected breast cancer cases and 387 controls were extracted from whole blood and genotyped for rs8176318 in the 3'UTR of the BRCA1 oncogene. The association with disease specific parameters and outcome were evaluated.

Results: Overall, there was a significant difference in the distribution of the three genotypes between cases and controls (p=0.035). The dominant variant model was predictive of breast cancer (OR=1.4, 95% CI 1.1−1.8). Fifty-two percent of breast cancer cases had the variant, with similar prevalence between subtypes: Luminal A (279 [54%] of 519 cases), Luminal B (37 [43%] of 85 cases), HER2 (21 [53%] of 40 cases) and triple negative breast cancer (41 [49%] of 83 cases). Comparing the prevalence of the variant within respective subtypes with controls however, Luminal A breast cancer was most strongly associated with rs8176318 (OR=1.5, 95%CI 1.1−1.9). The variant was not significantly associated with disease free survival (DFS) in all cancer cases (Log-rank test=0.084). However, rs8176318 was predictive of a poorer DFS (Log-rank test=0.041) in Luminal A patients with a 10-year DFS of 54% (95% CI 0.4−0.7) and 77% (95%CI 0.7−0.9) for the variant versus the non-variant, respectively. In addition, patients with Stage IV disease had a 6-fold increased risk of carrying the variant (p = 0.035), with 17 (73%) of 23 patients with metastasis at presentation positive for the variant compared to 324 (50%) of 642 patients without metastasis (p=0.034). Luminal A patients with Stage IV disease had a 13-fold risk of carrying the variant (p=0.043) in regression analysis, controlling for all other clinicopathological variables. Similarly, the variant was associated with distant metastasis in Luminal A patients, with 11 (92%) of 12 patients with metastasis positive for the variant compared to 242 (52%) of 463 patients without metastasis (p=0.028).

Conclusion: These findings suggest that rs8176318, a variant in the 3'UTR of BRCA1, is a genetic marker for modest breast cancer risk but aggressive tumor biology in breast cancer, and highlights the need for further clinical and biological evaluation of such markers.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-09-05.
P1-09-08: Association of Hypoxia-Inducible Factor-1 with Breast Cancer Risk: A Meta-Analysis of Published Studies.
Authors: Yin W , Liu G , Lu J , Shen Z , Shao Z
Published: June 2012

Background: Hypoxia-inducible factor-1 (HIF-1) plays an important role in the development and progression of breast cancer. However, conflicting results have been yielded for the association between HIF-1 polymorphisms and breast cancer risk. Therefore, we carried out a meta-analysis to assess the association of HIF-1 1772 C/T and 1790 G/A polymorphisms with breast cancer.

Material and methods: Computerized and manual searches were performed to identify eligible studies comparing allelic and genotypic frequency between cases and controls in breast cancer. Odds ratios (ORs) were used to estimate the association between HIF-1 polymorphisms and breast cancer risk. The fixed-effects or random-effects model was used to combine data. Subgroup analyses by ethnicity were also performed.

Results: This meta-analysis for 1772 C/T polymorphism included 1934 breast cancer cases and 1848 controls, and for 1790 G/A polymorphism included 602 breast cancer cases and 479 controls. We found that 1772C/T polymorphism increased the risk of breast cancer in the recessive model (OR = 2.273, 95% CI: 1.061−4.872, P = 0.035), whereas similar effect failed to be observed in the dominant model (OR = 1.075, 95% CI: 0.717−1.613, P = 0.725). There was a significant increase in breast cancer risk among Asian women (OR = 4.162, 95% CI: 1.508−11.484, P = 0.006) but not Turkish patients in the recessive model. For the 1790G/A polymorphism, we found that it had no exact effect on the decreased risk of breast cancer in both recessive and dominant model.

Discussion: This meta-analysis indicates that the polymorphism of HIF-1 1772C/T rather than 1790G/A polymorphism might be one of the high risk factors for breast cancer. The effect of the 1772C/T polymorphism on breast cancer risk is completely generated by the Asian women.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-09-08.
P1-11-04: Breast Cancer Treatment Resources and Guideline-Concordant Treatment in Appalachia.
Authors: Yao N , Hillemeier MM , Anderson RT , Matthews SA
Published: June 2012

Background: Appalachia has poorer cancer outcomes, but little research has been done regarding availability of cancer care resources in this region and how resource availability may relate to cancer outcomes. This study 1) examines associations between radiation therapy resources and receipt of radiotherapy after BCS in counties within Kentucky — a SEER state; and 2) describes spatial patterning of breast cancer treatment resources in all 13 Appalachian states.

Methods: For the Kentucky analyses, county-level data from the Area Resource File and SEER registry are analyzed. Bivariate analyses and spatial lag regression using a 6-nearest neighboring counties matrix are conducted. The sample includes stage I or II primary breast cancer patients age 18+ years diagnosed in Kentucky during 2000–2007. The dependent variable is the county-level percentage of patients received BCS without radiation; independent variables include density of radiation therapy providers and facilities and other socioeconomic covariates. For the analyses of entire Appalachian region, descriptive analyses and exploratory spatial data analysis are conducted including 420 Appalachian counties and 644 non-Appalachian counties in 13 states.

Results: In Kentucky 16.44% of 17,227 early stage breast cancer patients received BCS without radiation therapy (21.08% in Appalachia versus 14.80% in non-Appalachia, p<0.001). Appalachian Kentucky had significantly fewer radiation oncologists and radiation therapy facilities per capita than non-Appalachian Kentucky. The number of radiation therapy facilities per capita is negatively associated with rates of BCS without radiation when controlling for covariates. Analysis of 13 Appalachian states shows that Appalachian counties, especially in the Central and Southern regions, had significant fewer physicians per capita in Surgery, Anesthesia, Clinical Pathology, and Radiation Oncology. Clustering of scarce breast cancer care resources was observed in Central Appalachia.

Conclusions: Appalachian counties, especially in central Appalachia, have fewer breast cancer treatment resources than non-Appalachian counties, and resource availability is associated with cancer health disparities.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-04.
P1-11-12: Patterns of Care of Newly Diagnosed Patients with Breast Cancer in Mexico.
Authors: Chavarri-Guerra Y , Liedke PER , Symecko H , Hammond EE , Higgins MJ , Finkelstein D , Goss PE
Published: June 2012

Background: Breast cancer is the most common form of cancer and the leading cause of cancer death in women worldwide. More than 55% of breast cancer deaths occur in low and middle income countries. Although incidence rates for breast cancer are lower in developing countries, mortality rates are higher. This phenomenon has been attributed to limited access to care for breast cancer patients, including screening and early diagnosis as well as primary surgical, radiation and systemic therapies. Similar to trends in other poor and middle income countries, breast cancer mortality in Mexico is rising. The goal of this survey of physicians caring for patients with breast cancer in Mexico is to obtain information about current treatment patterns of newly diagnosed patients and to describe their clinical characteristics. Methods: A web-based closed survey has been sent to 854 physicians providing care to newly diagnosed breast cancer patients across Mexico, including medical oncologists and breast cancer surgeons. The survey instrument contains 35 questions assessing demographic data, access to diagnosis and treatment in a variety of clinical patient scenarios. The responses will be anonymous and entered automatically into a secure database for analysis. Fisher exact test will be used for the frequency analysis. Chi-squared statistics and Kendall correlation will be used for nominal and ordinal variables respectively. Results: The results will be presented at the 2011 San Antonio Breast Cancer Symposium.

Conclusions: The results of this survey will highlight potential disparities in care received by breast cancer patients across the full geographic and socioeconomic spectrum of Mexico in order to highlight the need for uniform, quality based approaches for the diagnosis and treatment of breast cancer patients in Mexico, and will serve as an example of how one middle income country faces challenges and unmet medical needs regarding access to care of women with breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-12.
P1-12-23: HER4 Coexpression Is Associated with Improved Recurrence Free Survival in HER2−Positive, Herceptin Treated Patients.
Authors: Brockhoff G , Machleidt A , Piendl G , Seegers S , Sassen A , Diermeier-Daucher S , Buchholz S , Ortmann O
Published: June 2012

Background: Individual therapy efficiency of HER2−positive metastatic and pre-metastatic breast cancer patients varies significantly and spans from effectual responsiveness over acquired insensitivity to complete resistance from the outset. Thus no predictive information can be deduced from HER2 diagnostics so that molecular biomarkers indicative for sensitivity / resistance to Herceptin are needed to be identified. The HER2 related HER4-receptor has been shown to have ambivalent (pro-apoptotic or pro-proliferative) activity and consequently represents a prime candidate to affect HER2 activity under Herceptin treatment. We retrospectively analyzed potential her4 gene amplification and HER4 protein expression in HER2−positive, Herceptin treated patients. Patient's overall and recurrence free survival was evaluated as a function of HER2/HER4 expression. Methods: Using dual color Fluorescence in-situ Hybridization (FISH probes, Zytovision, Bremerhaven, Germany) and qPCR (LC480, Roche, Penzberg, Germany) we quantitatively investigated primary breast cancer tissues from nearly 50 (FISH) and 160 (PCR) patients who received Herceptin treatment. We quantified the her4 gene copy numbers and evaluated the protein expression profile of all four known HER4 isotypes (JM-a/CYT1, JM-a/CYT2, JM-b/CYT1, JM-b/CYT2). Results: FISH analysis revealed a positive and independent prognostic marker in Herceptin treated breast cancer patients with respect to overall survival. Moreover by quantitative PCR analysis we found a significant variability of HER4 protein expression (JM-a/CYT1 and JM-a/CYT2; no JM-b isotypes) in HER2 positive breast cancer tissues, whereas HER2/HER4 positive patients show a significant better recurrence free survival compared to HER2 positive but HER4 negative patients (p = 0,003).

Conclusions: HER4 has been demonstrated to potentially exert tumor suppressing activity and in turn to have a favourable impact on the course of breast cancer disease. We show here that HER4 expression prolongs in particular recurrence free survival of Herceptin treated patients which indicates a functional integration of HER4 into anti-HER2 targeting. Complementing functional studies allowing for isotype specific function of HER4 will elucidate the special role of this receptor tyrosine kinase in the context of Herceptin treatment and might facilitate individualized anti-ErbB-receptor targeting with higher efficiency.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-23.
P2-01-10: Blockage of Autocrine TGF-b Signaling Reduces Tumorigenecity and Lung Metastasis in a Murine Breast Cancer Cell Line, NMuMG-ST.
Authors: Liu Z , Robert NW , Wang L , Bandyopadhyay A , Sun L-Z
Published: June 2012

Transforming growth factor beta (TGF-b) is known to have a dualistic role in breast cancer cells, acting as a tumor suppressor or a tumor promoter. This dualistic property varies in different cells and at different progression stages. Studies show that autocrine TGF-b is necessary for the growth and survival of MDA-MB-231 cells, and was also known to enhance the survival of MCF-7 cells. However, the role of autocrine TGF-b in spontaneously transformed cells is still less understood. We have retrovirally transduced a dominant negative TGF-b Type II receptor (DN RII) into a spontaneously transformed murine mammary gland epithelial cell line NMuMG-ST (DN RII cells). The expression of DN RII reduces TGF-b sensitivity of NMuMG-ST cells in a gene transcription assay and a growth inhibition assay. Interestingly, the autocrine TGF-b supports the survival of NMuMG-ST cells, although the exogenous TGF-b inhibits the growth of NMuMG-ST cells. Moreover, more apoptosis were observed in the DN RII cells compared to the control cells in both in vivo and in vitro experiment. We found that AKT and ERK pathway mediates autocrine TGF-b's survival signal. Results of western immunoblot for several stem cell markers and mammosphere formation assay suggest that autocrine TGF-b signaling is essential for the maintenance of stem like cell subpopulation in NMuMG-ST cells. By using immunocytostaining, we found that the DN RII cells expressed more E-cadherin and less Vimentin when compared to the control NMuMG-ST cells, which indicate that blocking autocrine TGF-b signaling pathway could inhibit the process of EMT in NMuMG-ST cells. Notably, when the cells were inoculated orthotopically in nude mouse mammary glands, tumors formed by DN RII cells grew at a similar rate as those formed by control cells, but caused fewer lung metastases. Our finding identified that the autocrine TGF-b signaling is important for the survival and metastatic ability of NMuMG-ST, which could provide an important foundation for further investigation on the role of autocrine TGF-b signaling in breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-10.
P2-01-24: Triple-Negative Breast Cancer Emerges from the Luminal Progenitor Compartment.
Authors: Nogi H , Kamio M , Kato K , Kawaase K , Toriumi Y , Takeyama H , Uchida K
Published: June 2012

Background: Triple-negative breast cancers (TNBC) are defined as tumors that lack expression of estrogen receptor (ER), progesterone receptor (PgR), and human epidermal growth factor receptor type 2 (HER2). Majority of TNBC (approximately 80%) are basal-like breast cancers. The question of whether there is a specific, identifiable cell in the normal breast form which basal-like breast cancers arise is controversial. We evaluated the frequency and receptor status of ductal carcinoma in situ (DCIS) and ductal components associated with T1 invasive ductal carcinoma (IDC) to clarify the developmental pathway of TNBC.

Methods: From our institutional archives between April 2000 and April 2011, we retrieved 758 cases of DCIS and T1 invasive ductal carcinoma without prior operative treatment. Immunohistochemistry for ER, PgR and HER2 was performed on ductal components and invasive components. Statistical analysis was performed using Fisher Extract Test.

Results: The frequencies of triple-negative (TN) subtype at each size were not seen significant differences. Ductal components were seen in 51 (84%) TN-IDC cases. Within 51 cases, the ductal components were TN in 49 cases. The remaining 3 cases were positive for ER and PgR, but negative HER2.

Conclusion: The resemblance between ductal components and invasive components and the coincident frequency of TNBC at each size sustain the hypothesis that TNBC arise from the luminal progenitor compartment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-24.
P2-02-03: MAP3K3, Amplified in Human Breast Cancer, Promotes Breast Tumor Progression and Defines Poor Prognosis.
Authors: Dong J
Published: June 2012

Gene amplification in the 17q chromosomal region is observed frequently in breast cancers. An integrative bioinformatics analysis nominated MAP3K3 gene, located in 17q23, as a potential therapeutic target in breast cancer. This gene encodes the mitogen-activated protein kinase kinase kinase 3 (MEKK3), but has not yet been associated with cancer-causal genetic aberrations. We found that MAP3K3 was amplified in approximately 8–20% of breast carcinomas, and that its over-expression was an independent prognostic marker for poor outcome with respect to relapse-free and overall survival, especially among the estrogen receptor-positive breast cancer patients. shRNA-mediated knockdown of MAP3K3 expression significantly inhibited cell proliferation and colony formation of MAP3K3-amplified breast cancer cell lines MCF7 and MDA-MB361, and promoted breast cancer cell apoptosis induced by TNFα, TRAIL, or a doxorubicin. In addition, ectopic expression of MAP3K3, in collaboration with Ras, induced colony formation in both primary mouse embryonic fibroblasts and immortalized mammary epithelial cells (MCF-10A). Together, these results suggest that MAP3K3 is a potential biomarker indicating poor prognosis, contributes to resistance to therapy, and is an oncogene in breast carcinogenesis. Therefore, therapeutic targeting of MAP3K3 may be attractive in breast cancer patients with MAP3K3-amplified breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-02-03.
P2-02-04: Novel Functions of vps34 in Non-Transformed Epithelial Cells: Regulation of Cell Proliferation and Tumorigenesis.
Authors: Su H , Xu T , Ganapathy S , Yuan Z-M
Published: June 2012

Autophagic and endocytic pathways are tightly regulated membrane rearrangement processes that are crucial for homeostasis, development and disease. A list of proteins regulating these cellular processes show altered expression in cancers, suggesting derailed autophagy or endocytosis as an emerging feature of cancer. Class III phosphatidylinositol 3-kinase (PI3kinase) Vps34 regulates both autophagic and endocytic systems by recruiting proteins containing PtdIns3P binding domains. Recently, many new insights into the biology of class III PI3Kinase have been reported. Most of these studies, however, used mainly transformed cells. It is unclear whether class III PI3Kinase works similarly in non-transformed cells. In the recent study, we explored this by depleting both transformed and non-transformed mammary epithelial cell lines of Vps34 using shRNA. In consistent with previous finding, vps34-depleted cells are defect in formation of FYVE-GFP puncta, and showed altered early and recycling endosome dynamics. In a shark contrast to transformed cells where knockdown of Vps34 was associated with decreased growth, Vps34 deficiency in non-transformed cells resulted in a marked increase in cell proliferation. The deregulated autophagy function found in Vps34-depleted cells appeared to be not involved in cell growth regulation as silence of either Atg7 or Beclin1 expression did not affect the growth rate. On the other hand, reporter luciferase-based ten-pathway screens suggested an altered activity of the MAPK pathway. Indeed, biochemical analysis indicated a sustained ERK activation resulting from the impediment of endosome-mediated EGFR turnover in Vps34 deficient cells. A contribution of the endosomal pathway to Vps34-mediated regulation of cell proliferation was further confirmed by using Rab5 and Rab11 mutants that deregulated early and recycling endosome, respectively. Importantly, inactivation of Vps34 compromised the cellular barrier against oncogene-mediated transformation, as demonstrated by the increased susceptibility of Vps34 depleted cells to Ras-induced colony formation in vitro and tumor development in mice. All together, our data uncover a novel role for the class III PI3Kinase-endosomal pathway in regulation of cell proliferation.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-02-04.
P2-02-08: Int6 Regulates Both Proteasomal Degradation and Translation Initiation and Is Critical for Proper Formation of Acini by Human Mammary Epithelium.
Authors: Suo J , Snider SJ , Lloyd RE , Chang EC
Published: June 2012

The mammalian INT6/EIF3E gene has been implicated in breast tumor formation, but its functional activities remain poorly defined. Consistent with the fact that Int6 is frequently down-regulated in human breast tumors, we found that repressing INT6 expression induced transformed properties in normal human mammary epithelial cells (MCF10A) in both 2D and 3D cultures. Int6 contains a PCI domain, found in several components of both the proteasome and the translation initiation factor eIF3, suggesting that it can functionally interact with both. Indeed, our data show that Int6 associates with proteasomes in human cells, and that INT6 knockdown prevents proper assembly of active proteasomes. In addition, we show that down-regulation of INT6 reduces translation initiation (both polysome formation and global protein synthesis). However, Int6 regulates translation selectively, reducing translation from cap-dependent and Bcl2- IRES reporters, but stimulating translation from the CVB3 IRES. These data collectively suggest that Int6 controls both protein synthesis and protein degradation, and is thus capable of fine-tuning protein levels. Abnormal levels of regulatory proteins caused by Int6 abnormalities may thus be responsible for disrupting normal morphogenesis of mammary epithelial cells and causing transformed phenotypes.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-02-08.
P2-05-05: Receptor-Like Protein Tyrosine Phosphatase Kappa (PTPRK) and Its Biological Role in Angiogenesis.
Authors: Sun P-H , Ye L , Mason MD , Jiang WG
Published: June 2012

Introduction: Protein Tyrosine Phosphatases (PTPs) are known as signalling molecules which affect cell growth, differentiation and oncogenic transformation. PTPs have been indicated in tumourigenesis and progression of various solid tumours. PTPRK, receptor-type protein tyrosine phosphatase kappa, has been shown to down-regulate transcriptional activity of beta-catenin and impact distribution of beta-catenin/E-cadherin complexes on cancer cell membrane. However, the role played by PTPRK in angiogenesis remains unknown. In present study, the effect of PTPRK knock-down on functions and tubule formation ability of HECV cells was investigated. (or In the present study, the effect of PTPRK on angiogenesis process was investigated.)

Methods: anti-PTPRK ribozyme transgenes were constructed to knock-down PTPRK expression in vascular endothelial cells, HECV cells. The subsequent effect upon in vitro cell growth, adhesion, migration and microvascular tubule formation was examined using a variety of functional assays.

Result: Knock-down PTPRK in HECV cells (HECVΔPTPRK) resulted in a decrease of cell growth in vitro. The growth rate of HECVΔPTPRK was 276.3±16.4, p=0.01 compared with HECVpEF (314.8±21.9) controls. However, knock-down of PTPRK in HECV cells increased cell motility. The cell migration distance of HECVΔPTPRK was 83.8±19.8μm, p=0.008 compared with HECVpEF (61.8±11.8μm) controls. No effect on cell adhesion by PTPRK knockdown was seen in HECVΔPTPRK (81.2±6.6), compared with HECVpEF control (78.8±15.6). Furthermore, knock-down of PTPRK suppressed tubule formation in HECV cells, the length of total tubules (μm) in HECVΔPTPRK was 2167.4±943.2, p<0.05 compared with HECVpEF (3130.6±386.9) control.

Conclusion: Knock-down of PTPRK reduced the growth and tubule formation abilities of vascular endothelial cells. It suggests that PTPRK is pivotal regulator for angiogenic process. Further investigations are required to identify the downstream pathways involved in these impacts.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-05-05.
P2-05-06: Role of Repulsive Guidance Molecule b (RGMb) in HGF Mediated Angiogenesis.
Authors: Sanders AJ , Ye L , Mason MD , Jiang WG
Published: June 2012

Introduction: Hepatocyte Growth Factor (HGF) has been widely documented as playing a key role in enhancing the aggressive nature of cancer through its ability to promote cellular processes such as migration, invasion and angiogenesis. Development of a blood supply is vital to advanced tumour growth and increased metastatic potential. In the current study we identified RGMb (DRAGON), a member of the Repulsive Guidance Molecule family, as being upregulated by HGF and assessed its potential to contribute to HGF mediated proangiogenic traits.

Methods: Expression of RGMb was identified as being upregulated in human endothelial HECV cells following 4 hour treatment with HGF (40ng/ml) using micro array analysis and Q-PCR. Subsequently, the expression of RGMb was targeted through the transfection of HECV cells with a plasmid containing a ribozyme transgene specifically targeted to RGMb. The role of RGMb in HGF mediated cellular migration and tubule formation in vitro was examined.

Results: Significant increases in RGMb expression in HECV cells were observed on the mirco-array following 4 hour treatment with HGF (P = 0.004) and this trend was also identified using Q-PCR. Transfection with the ribozyme transgene brought about substantial reductions in RGMb expression at both transcript and protein levels as assessed using RT-PCR, QPCR and Western blot analysis. Knockout of RGMb brought about a significant increase in migration rates, compared to HECV cells transfected with a closed pEF6 plasmid only (HECVpEF6), following a 90minute period (p = 0.034 at 90 minute time point vs pEF6 control). Treatment of HGF enhanced migration rates of HECV control cells with significant differences between untreated and HGF treated pEF6 cells obvious following 60 minutes (p < 0.05 at 60 and 75 minutes, p = 0.002 at 90 minutes). In contrast to this, HECV cells transfected with the RGMb ribozyme transgene (HECVRGMbKO) were unaffected by HGF treatment, with no significant differences observed between treated or untreated HECVRGMbKO cells at any time points. A similar trend was observed in the angiogenic tubule formation assay, where treatment of HECVpEF6 cells with HGF could significantly enhance the levels of tubules formed (mean tubule perimeter, untreated 8087+/− 632 vs HGF treated 13131+/− 988, p = 0.001). Treatment of HECVRGMbKO cells with HGF did not significantly enhance levels of tubule formation (mean tubule perimeter, untreated 7523+/−2458 vs HGF treated 11050+/−1512, p = 0.21).

Conclusions: Targeting of RGMb in endothelial cells appears to reduce their sensitivity to the promotional effect of HGF on cell migration and tubule formation, important traits in the angiogenic cascade. Our data suggests that RGMb may be one molecule involved in the process through which HGF enhances angiogenic potential and targeting this molecule may be a useful strategy in a number of cancer types to interfere with HGF promoted angiogenesis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-05-06.
P2-05-10: Brain-Derived Neurotrophic Factor, BDNF, and Its Biological Impact on Vascular Endothelial Cells.
Authors: Jiang WG , Ye L , Yang X , Mansel RE
Published: June 2012

Background: BDNF, Brain-derived neurotrophic factor is a member of the neurotrophin superfamily and is a known growth factor for both central and peripheral nervous systems. While BDNF has been shown to play a role in the development of a number clinical condition including epilepsy, Alzheimer's and Parkinson's disease, it has recently been implicated in human cancers, primarily in neurological related malignancies, as well as ovarian cancer, lung cancer and skin malignancies. We have recently reported a pivotal relationship between BDNF and disease progression and clinical outcome in human breast cancer. Certain members of the neurotrophin family have also been shown to play a role in the regulation of angiogenesis. In the present study, we examined the impact of BDNF on the biological behaviour of vascular endothelia cells.

Materials and methods. Human vascular endothelial cells were used in the present study. A human BDNF expression plasmid was constructed from a cDNA library of normal tissues. Anti-BDNF transgenes were constructed based on the secondary structure of human BDNF and used to knockdown the expression of BDNF from the cells. Vascular endothelial cells with differential expression of BDNF were generated using the expression construct. Cell-matrix adhesion, cell migration and growth were determined.

Results. Human vascular endothelial cells (HECV) expressed very low levels of BDNF, traceable by PCR method. The cells were transfected with BDNF expression plasmid, which allowed establishment of BDNF over-expressing sublines, HECVBDNFexp. Expression of BDNF had a marked influence on matrix adhesion of the endothelial cells (adhesion indices for control and HECVBDNFexp cells being 3.3± 0.83 and 2.18± 0.94, respectively, p=0.01). It is interesting to note that the change of adhesion was likely to involve the focal adhesion (FAK) pathway as a FAK inhibitor, PF573228, had a profound impact on the adhesion of both control and HECVBDNFexp cells. Manipulation of BDNF in endothelial cells had a similar effect on the cellular migration, although to a lesser degree. Conclusions. Brain-derived neurotrophic factor, BDNF, has a profound impact on the biological functions of vascular endothelial cells. This may have implications in the angiogenic process seen in solid tumours.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-05-10.
P2-09-08: FDG-PET in the Staging of the Axilla in Women with Breast Cancer Treated with Primary Chemotherapy.
Authors: Lao Romera J , Puértolas Hernandez T , Alvarez Alejandro M , Godoy Molias A , Hernandez Garcia A , Millastre Bocos E , Garcia Mur C , Ubieto Laguarda MA , Artal Cortes A , Anton Torres A
Published: June 2012

Background

FDG-PET has proven to be useful during the initial staging of several tumor types. The aim of the present study was to assess the accuracy of this test in the assessment of initial axillary status in patients referred to treatment with induction chemotherapy.

Material and Methods: Women with stage II-IIIA (tumors measuring ≥3cm or positive-staged axillary involvement) to be treated with neoadjuvant Docetaxel-Doxorubicin-Cyclophosphamide (TAC scheme) were included. In every patient a physical exam, MRI, ultrasound-guided FNAB and FDG-PET were performed prior starting chemotherapy. The criteria to consider the axilla to be initially positive were positive surgical staging or type-D pathological response (Miller-Payne) in the axilla. The ability of FDG-PET to predict pathological response in patients with breast cancer was assessed and compared with other routine procedures.

Results: 42 patients (p) were included. Median age was 45 years (30-66). 11p (26.2%) were triple-negative. 2p (4.8%) were Her-2 +. By clinical stage, 13p (30.9%) were IIA, 17p (40.4%) IIB and 12p (28.2%) IIIA. 41p (97.6%) underwent surgery after induction chemotherapy. According to surgical staging 31p (75.6%) were deemed to be N+ from the diagnosis (19p with pathologic N1-3a and 12p with type-D nodal response). PET scan positively detected deposits in the axilla in 26p (83.9%) of those patients compared to 22p (70.9%) with ultrasound-FNAB, 19p (61.3%) physical exam and 14p (45.2%) MRI.

Conclusions: In this study FDG-PET could predict axillary involvement more precisely than other routine staging procedures in these patients submitted to be treated with induction chemotherapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-09-08.