bluuebunny/crossref_metadata_2025 huggingface dataset:
Correlation of Thymidine Phosphorylase Expression and Lymphocyte Infiltration Detected by Microarray Analysis of Breast Cancer.
Authors: Ruckhäberle E. , Rody A. , Holtrich U. , Engels K. , Gaetje R. , Turley H. , Hanker L. , Solbach C. , Karn T. , Kaufmann M.
Published: December 2010

Background:Thymidine phosphorylase (TP) expression in carcinoma cells has been described as a prognostic factor in breast cancer and a predictive factor for response to 5-FU chemotherapy in some studies. Analysis from prostate cancer reported a correlation of TP with lymphocyte infiltrates of tumors.Material and Methods:Affymetrix microarray data of n=79 normal tissue samples and n=1781 primary breast cancers were analyzed for TP mRNA expression. To study relationships of TP expression with immune cell infiltration of breast tumors we used several metagenes representing certain types of immune cells. Immunohistochemical analysis of TP protein expression using a monoclonal antibody was applied for validation studies.Results:Among normal tissues highest expression of TP mRNA was observed in cells and tissues of the immune system. The profile of TP expression displayed highest correlation with a metagene representing cells of the myeloid lineage as moncytes, macrophages and dendritic cells. Analysis of microarray data from 1781 breast cancer samples suggests that TP expression detected by this method originates mainly from infiltrating immune cells. In line with this observation TP mRNA expression correlated with a immune cell infiltration score determined by pathological inspection of the tumor. However, we also observed a correlation of TP with a metagene of interferone inducible genes which seem to be expressed by carcinoma cells. When we tried to validate these data on the protein level using immunohistochemistry TP expression was demonstrated both in carcinoma cells and stromal cells of the tumor to a varying degree.Discussion:We were able to confirm previous data from prostate cancer that TP expression is strongly correlated to the presence of an intense lymphocyte infiltrate of the tumor for breast cancer. However, microarray data of a bulk tumor sample cannot reveal the cellular origin of TP expression.Conclusion:Thus from this data it is not clear whether lymphocytes of the myeloid lineage either are the actual source of TP expression or induce its expression in carcinoma cells.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2139.
NF-kB Complex Activation and Association of αB-Crystallin and Vimentin with Poor Prognosis in Primary Breast Cancer.
Authors: Sreseli R. , Quinlan P. , Quinlan R. , Bray S. , Kellok D. , Baker L. , Jordan L. , Purdie C. , Thompson A.
Published: December 2010

Background: Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) is a protein complex found in almost all animal cell types and is involved in cellular responses to stimuli such as stress, free radicals, cytokines et.c. NF-kB plays a key role in regulating the immune response and incorrect regulation of NF-kB has been linked to the development of cancer, several inflammatory and autoimmune diseases. The role of NF-kB, its subunits and pathways in breast cancer and in the surrounding stromal tissues remains uncertain. αB-crystallin is a heat shock protein, which function as stress-induced molecular chaperones to inhibit the aggregation of denatured proteins. Previous studies have identified αB-crystallin as a marker of poor prognosis for breast cancer and have suggested that it is an excellent marker for tumours of basal origin. The aim of this study is to examine, for the first time, NF-kB and αB-crystallin, vimentin and HSPs in the same set of breast cancers and stroma and relate this to clinical parameters including outcome.Methods: Tissue Micro Arrays of 0.6mm cores of 246 breast cancers were stained with antibodies to αB-crystallin, vimentin, HSP27 (antibody ERD5) and HSP27 82P and 12 subsequent antibodies for NF-kB (IKKa, Ser276, Ser32/36, 180/181, P52, 1kkygmma, P65, RELB, P100/P52, IkBa, cREL, and P50) and scored using the Quick Score Method.Results: Positive associations have been found between αB-crystallin and IKBA (P=0.001 Fishers exact test (FET)), RELB (P<0.001 (FET)) and SER32/36 (P=0.03 (FET)) whereas Vimentin was positively associated IKBA (P=0.003 (FET)), IKKA (P=0.001 (FET)), P50 (P=0.002 (FET)), RELB (P<0.001 (FET)) and SER32/36 (P=0.01 (FET)). Positive associations have also been found between HSP27 and P50 (P=0.002 (FET)), SER32/36 (P<0.001 (FET)). Antibodies such as 180/181 (P=0.04 Kaplan Meier Log Rank (KM), cREL (P=0.03 (KM)), IKBA (P=0.03 (KM)) and IKKA (P=0.01 (KM)) was associated with good survival and P50 (P=0.002 (KM)) and P65 (P=0.01 (KM)) to poor survival. Expression of αB-crystallin was associated with vimentin (P<0.001 (FET)). Its expression was linked to a low expression of the estrogen receptor. Vimentin expression was associated with estrogen receptor (ER) negative cancers and poor survival (P <0.001 (FET)), P=0.002 (KM Log Rank) respectively). In contrast to αB-crystallin, low expression of HSP27 was associated with low ER and progesterone receptor (PGR).Conclusions: Associations were found between various antibodies within the NF-kB pathway as well as the heat shock proteins αB-crystallin, vimentin and HSP27 with varying implications for overall survival. We aim to clarify the mechanisms to prevent the aggregation of stress-accumulated misfolded proteins and apoptosis resulting better survival.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2143.
Regulators of Mitotic Transition Determine Paclitaxel Resistance In Vitro and Predict Poor Prognosis in Breast Cancer In Vivo.
Authors: Burrell R. , McClelland S. , Juul N. , Downward J. , Szallasi Z. , Szallasi Z. , Swanton C. , Swanton C.
Published: December 2010

We have previously identified 25 kinases that regulate sensitivity to the microtubule-stabilising drug, paclitaxel (Swanton et al., 2007). Silencing these kinases by small-interfering RNA (siRNA), results in resistance to paclitaxel-induced cytotoxicity and dramatic alterations in DNA ploidy, multinucleation and centrosome amplification independent of drug treatment. These data suggest that the molecular pathways that maintain chromosome stability and mitotic fidelity are required for paclitaxel sensitivity. To further address this, we have investigated the function of the 25 kinases in mitosis, using a combination of fixed and live-cell microscopy. Several of these genes regulate mitotic progression and the formation of a bipolar mitotic spindle, and upon depletion, lead to increased rates of chromosome missegregation during anaphase. This supports the hypothesis that mechanisms of chromosomal instability that are common in cancer cells, predispose to paclitaxel resistance. In order to assess the in vivo relevance of these kinases we examined whether lower expression held prognostic power across several breast cancer datasets. Notably low expression of MAST3, CAMK1 and SKAP1 correlates with poor prognosis in breast cancer in vivo (P<0.05). Interestingly, MAST3 and CAMK1 have strikingly similar mitotic phenotypes after silencing; cells arrest in early mitosis and show an increased frequency of anaphase errors that may contribute to chromosomal instability. In summary, we have identified novel regulators of mitotic transition that may be important for maintaining chromosomal stability. These kinases influence taxane sensitivity in vitro and hold prognostic power in breast cancer patients in vivo.ReferencesSwanton, C., Marani, M., Pardo, O., Warne, P.H., Kelly, G., Sahai, E., Elustondo, F., Chang, J., Temple, J., Ahmed, A.A., et al. (2007). Regulators of mitotic arrest and ceramide metabolism are determinants of sensitivity to paclitaxel and other chemotherapeutic drugs. Cancer Cell 11, 498-512

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2151.
The Hepatitis A Virus Cellular Receptor, HAVcR-1, Reduces the Integrity of Human Endothelial Tight Junctions.
Authors: Martin T. , Harrison G. , Mason M. , Jiang W.
Published: December 2010

Background: Disruption of Tight Junctions (TJ's) in endothelial cells can cause a leaky vascular bed and may therefore lead to vascular spread of cancer cells. Hepatitis A virus (HAV) cellular receptor HAVcR-1, also known as KIM-1/TIM-1, is the cellular receptor for the hepatotropic picornavirus that causes acute hepatitis-A in humans. Although HAVcR-1 is expressed in every human organ, the natural function of HAVcR-1 remains unknown. In the current study, we investigated the location, association and possible functionality of HAVcR-1 in human endothelial cells.Methods: HAVcR-1 protein in human endothelial cells was visualised using immunohistochemistry and mRNA transcript by RT-PCR. A hammerhead ribozyme transgene that specifically targeted human HAVcR-1 was constructed and used to knockdown endogenously expressed HAVcR-1 (HECVHKD); moreover, forced expression was obtained by insertion of a transgene into wild type endothelial cells (HECVHEX).TJ function was assessed using trans-endothelial resistance (TER) and paracellular permeability assays under the influence of HGF (40ng/ml) a cytokine we have previously shown to modulate TJs in human endothelial cells.Results: Human endothelial cells expressed HAVcR-1 at low levels. The location of both endogenous and forcibly expressed HAVcR-1 was at the cell-cell junction, at the region of the TJ. In this study, one of the first to examine the location and binding partners of HAVcR-1, expression of this receptor was targeted to the vicinity of intercellular junctions, via ZO-1, which further was demonstrated to be at the site of the TJ by its co-localisation with ZO-2. This was true of both endogenously and forcibly expressed protein. Moreover, HAVcR-1 was co-precipitated with the regulatory factor Rho C. Although there was no change in function of HECVHEX cells, the TJ function of the HECVHKD cells exhibited significantly reduced response to HGF over 2h (p<0.01). Interestingly, there was no concurrent response in PCP.Conclusion: These results demonstrate for the first time that HAVcR-1 may have a previously undiscovered role in the regulation of TJ integrity in human endothelial cells.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2158.
ER and PR Expression Profile of Circulating Tumor Cells in Metastatic Breast Cancer Patients in Comparison to the Primary Tumor.
Authors: Aktas B. , Mueller V. , Schumacher K. , Tewes M. , Zeitz J. , Kasimir-Bauer S. , Rack B. , Janni W. , Solomayer E. , Fehm T.
Published: December 2010

Background: Introduktion: Several studies have indicated that the expression of predictive markers including ER and PR can change during course of disease. Therefoore, reassesment of of the prediktive markers at the time of disease progression might help to optimize treatment decisions. Metastatic tissue may be difficult to obtain for repeated analysis. In this context, characterization of circulating tumor cells (CTCs) could be of relevance. Therefore, the purpose of the present study was: (1) to reevaluate the ER/PR expression by circulating tumor cells and (2) to compare the hormone receptor status expression profile of CTC with the primary tumor. Materials and Methods: We ecaluated, 166 blood samples from metastatic breast cancer patients at the time of first diagnosis of first metastatic disease or disease progression. All samples underwent immunomagnetic enrichment using the AdnaTest BreastCancerSelect (AdnaGen AG, Germany) within 4 hours after blood withdrawal followed by RNA isolation and subsequent gene expression analysis by reverse transcription and Multiplex-PCR in separated tumor cells using the AdnaTest BreastCancerDetect. CTC were analyzed for the three breast cancer associated markers: GA733-2, Muc-1, Her-2 and β-actin as an internal PCR control. Expression of the estrogen (ER) and progesterone (PR) receptor was assessed in an additional RT-PCR. The analysis of PCR products was performed by capillary electrophoresis on the Agilent Bioanalyzer 2100. Results: The overall detection rate for CTC was 38% (63/166 patients) with the expression rates of 79% for EpCAM (50/63 patients), 77% for MUC1 (49/63 patients), 49% for HER2 (31/63 patients), 23% for ER (15/65 patients) and 11% for PR (7/65 patients), respectively. Comparisons with the primary tumor were only performed in CTC+ patients (n=65). In 41 of 65 (63%) patients with ER+ tumors, CTC were ER- and 38/65 (58%) patients with PR+ tumors did not express PR on CTC. Primary tumors and CTC displayed a concordant ER and PR status in only 34% and 61% of cases, respectively. Conclusion: Most of the CTC were ER/PR-negative despite the presence of an ER/PR-positive primary tumor. The predictive value of hormone receptor status expression profile of CTC for palliative endocrine therapy has to be prospectively evaluated.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3009.
Circulating Epithelial Cells as a Tool for Monitoring Treatment Success of Primary Chemotherapy with Simultaneous or Sequential and without Trastuzumab.
Authors: Camara O. , Kroll T. , Oppel S. , Runnebaum I. , Pachmann K.
Published: December 2010

In spite of ample prognostic markers available in breast cancer, still a considerable proportion of patients with good prognostic markers suffers relapse whereas patients with poor prognostic markers may remain disease free. It would, therefore, be desirable to control, at the individual patient level, whether the applied therapy is effective. Our previous work indicates, that in cancer patients most of the epithelial cells circulating in peripheral blood (CETC) are part of the tumor and that the response of these cells reflects the response of the tumor to the applied therapies.Therefore monitoring the decrease or increase in numbers of these cells providing a unique tool for therapy surveillance was used to monitor neoadjuvant chemotherapy in 26 her2/neu positive breast cancer patients with either IHC3+ or FISH confirmed Her2/neu positive breast cancer. Patients were prospectively analysed for the number of CETC before therapy, before each new cycle of chemotherapy and during maintenance therapy at each visit initially every three months and subsequently at more extended intervals. 1ml of blood was drawn into EDTA vials, red blood cells lysed and the white blood cell pellet stained with FITC-labelled anti-Epcam. Green fluorescent cells were detected by image analysis and dead cells excluded due to red PI fluorescence.After an initial variable reduction of CETC during neoadjuvant chemotherapy, tightly connected to tumor reduction we regularly observed a massive release of cells from the shrinking tumor. Although these cells may not be able to settle in tumors with low metastatic potential, in 4/4 patients with Her2/neu positive tumors who did not receive trastuzumab the number of CETCs further increased after termination of therapy and surgery followed by rapid distant relapse indicating that the cells released in these patients may be highly aggressive with a high potential to settle and grow into metastases.It is, however not clear, whether sequential or simultaneous addition of trastuzumab to chemotherapy is preferential. In our hands 65% of the patients receiving simultaneous trastuzumab showed increasing CETCs and all have suffered relapse whereas all patients with decreasing CETCs are still in complete remission indicating that in the neoadjuvant situation during maintenance therapy an increase in CETC is the earliest indicator of imminent relapse.6/6 patients who received trastuzumab sequentially to the neoadjuvant treatment all showed decreasing numbers with all of them still without signs of disease after 4 years of follow up. Thus, trastuzumab was highly effective in this treatment and even if the tumor cells were not eliminated immediately, trastuzumab contributed to prevent them from settling and growing into metastases and CETC monitoring favours sequential trastuzumab.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3016.
Single Circulating Tumor Cell Detection and Overall Survival in Non Metastatic Breast Cancer.
Authors: Bidard F. , Mathiot C. , Mefti F. , Delaloge S. , Giacchetti S. , Salmon R. , Mignot L. , Sigal-Zafrani B. , Vincent-Salomon A. , Tembo O. , Marty M. , Pierga J.
Published: December 2010

Background: Circulation of cancer cells in the blood is a necessary step of hematogeneous metastasis while circulating tumor cells (CTC) have been reported to have a low metastatic efficiency in preclinical animal models. After a median follow-up of 18 months, we previously reported that CTC detection influences the distant metastasis-free survival (DMFS) in non-metastatic breast cancer (BC) patients (pts) treated by neoadjuvant chemotherapy (NACT) in a multicenter prospective trial. Updated results are presented here, focusing on overall survival (OS) and predictors of metastatic relapse.Methods: In 115 localized BC pts, CTC were prospectively screened (CellSearch) before and after NACT (REMAGUS02). We analyzed their outcome after a median follow-up of 36 months.Results: At baseline, 23% of pts were CTC-positive, but only 10% had more than 1 CTC per 7.5ml of blood. At an individual level, CTC detection before chemotherapy, used as a test to predict metastatic relapse, exhibited a global accuracy of 77%, higher than that of tumor grade (54%), tumor size (57%), lymph node invasion (40%), triple negative phenotype (76%) and pathological complete response (27%). Multivariate analyses for OS and DMFS showed that CTC detection before chemotherapy was a strong independent prognostic factor for both DMFS (p=0.01, RR=5.0, 95%CI[1.4-17]) and OS (p=0.007, RR=9, 95%CI[1.8-45]), along with tumor size and triple-negative phenotype, while post-chemotherapy CTC detection had a lower significance for both endpoints (p=0.07 and p=0.09 respectively).Conclusion: Biologically, the metastatic efficiency of CTC could be higher than previously thought. Clinically, besides confirming our previously reported results, this study shows that CTC detection may become the main prognostic factor in BC pts treated with NACT. Implementing this technique in everyday management might help to identify high-risk pts in whom innovative strategies should be investigated.Supported by PHRC AOM/2OO2/02117, Pfizer inc., Roche, sanofi-aventis.ISRCTN10059974

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3017.
Relevance of Traditional CEA and CA15-3 and Bone Turnover Markers in Predicting Bone Relapse in Breast Cancer Patients.
Authors: Ibrahim T. , Sacanna E. , Mercatali L. , Scarpi E. , Ricci M. , Ricci R. , Serra P. , Tison C. , Amadori D.
Published: December 2010

Background: Over 50% of breast cancer patients who relapse systemically develop bone metastases. The RANK/RANKL/OPG axis governs osteoclastogenesis and bone reabsorption. In particular, osteoprotegerin (OPG) is a decoy receptor of RANK-L (receptor activator of nuclear factor kappa B ligand) and seems to prevent bone destruction by blocking the binding of RANKL with its receptor. We studied the efficacy of circulating OPG compared to the traditional markers CEA and CA15-3 for the early detection of bone lesions.Methods: The study was carried out on peripheral blood from 72 patients with breast cancer, of whom 33 were disease-free and 39 had bone metastases. The OPG transcript was determined using quantitative PCR analysis, and the traditional markers were quantified by immunoenzymatic assays.Results: The OPG median value was higher in disease-free patients (median = 1.7, 0.4-8.9) than in bone-relapse patients (median = 0.6, 0.1-5.2) (p<0.001). CEA and CA15-3 median values were 1.3 (0.0-8.3) ng/ml and 10.6 (0-45.7) U/ml, respectively, in disease-free patients and 4.8 (0.0-90.6) ng/ml and 69.8 (7-1538) U/ml, respectively, in patients with bone metastases (p< 0.0001 for both markers). Considering 0.9 as cut off of OPG relative expression, our analyses revealed a specificity of 88% with respect to 97% for CEA and 94% for CA15-3. OPG sensitivity was 73% compared to only 49% for CEA and 67% for CA15-3. When OPG was considered in combination with the traditional markers, sensitivity increased from 49% to 83% for CEA and from 67% to 88% for CA15.3.Conclusions: Our results on a preliminary series of breast cancer patients indicate that the bone turnover marker OPG greatly increases CEA and CA15-3 sensitivity in predicting bone metastases.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3027.
Prospective Evaluation of Serum HER2 and HER2 Expression on Circulating Tumor Cells in Patients with Metastatic Breast Cancer within the DETECT Study.
Authors: Mueller V. , Riethdorf S. , Rack B. , Janni W. , Fasching P. , Pantel K. , Gebauser G. , Solomayer E. , Aktas B. , Kasemier-Bauer S. , Fehm T.
Published: December 2010

Background: Assessment of HER2 status at the time of metastatic disease might help to optimize treatment decisions in patients with no HER2 status determined at first diagnosis and could also be helpful since HER2 status may change during disease progression. In this context, determination of serum HER2 or the evaluation of HER2 status on circulating tumor cells (CTC) is of potential relevance especially for optimizing HER2-directed therapy. The aim of this study was therefore to determine the serum HER2 status and the HER2 status of CTC from corresponding patients.Methods: Blood samples were obtained in a prospective multicenter setting from 251 patients with metastatic breast cancer at the time of disease progression. Serum HER2 was determined using the commercial HER2 ELISA-kit (Oncogene Science). CTC were detected and HER2 expression was assessed with the CellSearch™ system (Veridex).Results: 119 of 251 (47%) metastatic patients had elevated serum HER2 levels above 15 ng/mL. Serum HER2 was elevated in 49 (66%) of 74 patients with HER2 positive primary tumors, in 57 of 148 (39%) with HER2 negative primary tumors and in 13 of 29 (45%) patients with unknown HER2 primary tumor status. CTC were detected in 133 of 243 evaluable patients (5 or more cells), in these HER2 overexpression on CTC (strong overexpression in at least one cell) was detected in 52 cases (39%). Only 20 of these patients had a HER2 positive primary tumor. In patients with HER2 positive CTC, serum levels were above the cut off in 34 of 52 (65%), while in those with HER2 negative CTC 44 of 81 (54%) showed elevated HER2 serum levels. Concordance between HER2 status of circulating tumor cells and elevated serum HER2 was seen in 71 of 133 patients (53%).Conclusions: Our study demonstrates with a prospective design that also in patients with initially negative HER2 tumor status elevated serum HER2 levels and/or HER2 positive CTC can be detected at the time of metastatic disease. This is of clinical relevance since these patients in current practice do not have access to HER2 targeted therapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3050.
Amplification of mRNA from Formalin-Fixed Paraffin-Embedded (FFPE) Breast Cancer Tissues without 3' Bias and Multiplex Expression Analysis on Flow-Through Microarrays.
Authors: Englert D. , Wilson D. , Laken S.
Published: December 2010

Background: FFPE samples are an abundant sample resource for validation of expression-based assays and are will suited for routine clinical testing. Analysis on microarrays enables the analysis of many transcripts from small samples. However, mRNA in FFPE sections is degraded and cross-linked, with RNA fragment sizes of a few hundred bases or less, and consequently standard methods of microarray sample preparation are not reliable unless probes are very strongly biased to the 3' end. It is not always possible to design high quality probes within this restricted region of mRNAs. Amplification of target sequences any place within transcripts would permit greater flexibility in probe design and optimization of probe sets for reliable multiplex analysis.Material and Methods: Total RNA extracted from FFPE samples was subjected to two rounds of amplification with the ExpressArt TRinucleotide kit (AmpTec), which preferentially primes cDNA synthesis near the 3'-ends of mRNAs (rather than at poly-A sites) and selects for mRNAs against ribosomal RNAs. Biotin-labeled anti-sense RNA (aRNA) was prepared from the second round cDNA with the Illumina® TotalPrep™ RNA kit (Applied Biosystems). The labeled aRNA was hybridized to probes immobilized in singlet on flow-through microarrays on the Ziplex Automated Workstation (Xceed Molecular). Performance was assessed by the analysis of titration mixtures prepared from mRNA from breast cancer and colon cancer FFPE tissue blocks.Results: Median CVs of technical replicates were less than 25%. Linear responses were observed in the analysis of the titration mixtures, and all correlations between predicted and observed expression values of the titration mixtures had R2 values greater than 0.8. The data indicated that the minimum detectable expression difference was less than two-fold. Similar signal intensities were observed for 3'-biased probes and probes several hundred bases from the 3'end, confirming the lack of 3' bias in the amplification.Discussion: The results demonstrate the feasibility of amplifying and quantifying sequences at any position within transcripts in degraded mRNA from FFPE samples. Expression differences of less than two-fold may be analyzed with tens or hundreds of probes for translational research and clinical assay development on the Ziplex Workstation. Many probes may be tested in parallel to optimize probe sets for specific tests.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3052.
Genetic Polymorphisms in EGFR Gene: No Evidence for an Association with the Risk of Breast Cancer.
Authors: Gorlov I. , Gorlova O. , Jupe E.
Published: December 2010

EGFR expression is an important predictor of prognosis for breast cancer. An association of the EGFR expression breast tumor with the prognosis suggest that germline polymorphisms in the gene also may be associated with the risk of breast cancer. The published studies on the association of the genetic polymorphisms in the EGFR gene are conflicting.We genotyped 15 SNPs in the EGFR gene region: rs11506105, rs11543848, rs12674036, rs1558544, rs2072454, rs2075109, rs4947986, rs6965469, rs6969537, rs759171, rs7801956, rs4947963, rs747322, rs17172451, and rs1554718 in 500 Caucasian breast cancer patients and 500 matched controls to assess the association of the SNPs with the risk of breast cancer. SNPs were selected based on the LD pattern in the region and how close they are located to exons and promoter. Additionally we analyzed an extremely polymorphic CA repeat (rs11568315) located in the regulatory region of the gene. This polymorphic repeat has 11 alleles (alleles were defined by the number of the CA repeats, 14-23).We conducted pooled analysis and also stratified subjects by menopausal status. We have not found any significant association of the polymorphism with breast cancer risk. We were not able to confirm the previous finding that the increased number of the CA repeats in the rs11568315 was associated with elevated breast cancer risk. In conclusion, despite the fact that the expression of EGFR in breast cancer is an important predictor of progression, we found no evidence that germline polymorphisms in the gene are associated with the increased risk of breast cancer. The study was supported by 1R03CA128025-01A1 grant.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3060.
Differences in Breast Cancer Subtypes among Asian-American Women with Invasive Breast Cancer in New York City.
Authors: McCarville K. , Flam A. , Forst M. , Swistel A. , Osborne M. , Moore A. , Vahdat L. , Klein P. , Christos P. , Mazumdar M. , Chuang E.
Published: December 2010

Background: Specific breast cancer (BC) subtypes conferring distinct phenotypic and prognostic outcomes have been defined by molecular gene expression arrays. A recent study suggested that differences in BC subtypes exist among ethnic subgroups of Asian women in California. It has also been reported that there are differences in clinical outcomes among Asians with BC in the US, with Filipinos having a worse prognosis compared with other Asians. We sought to determine whether differences in BC subtypes exist among Asians in New York.Methods: Using outpatient registration records from Weill Cornell Breast Center and St. Vincent's Medical Center in New York City, we identified patients who were diagnosed with stage I, II, or III invasive BC between 1997 and 2007 who were of Chinese, Filipino, Japanese or Korean ethnicity. We reviewed pathology records according to an IRB approved protocol and recorded patient data for age, stage, grade, estrogen receptor (ER), progesterone receptor (PR) status, and HER-2/neu at diagnosis. Immunohistochemical surrogates for the four molecularly defined breast cancer subtypes were used according to accepted definitions (Luminal A: ER+Her2-; Luminal B: ER+ Her2+; Her2/neu: Her2+ ER-; Basal-like: ER-PR-, Her2-) Using chi-square analysis, we explored the relationship between ethnicity and BC subtypes.Results: 346 Asian women were identified with invasive breast cancer. Among the Chinese subgroup, 67% were of the Luminal A subtype, 15.3% were Luminal B, 10% were the Her2/neu subtype and 8.5% were Basal-like. Filipinos had a higher proportion of Luminal B cancers compared to the other ethnic groups. Filipinos and Koreans had a higher proportion of the Her2+/ER- subtype compared to Chinese and Japanese (P=0.004 by chi-square test). When considering all Her2/neu positive cancers, Filipinos had a significantly higher proportion of HER-2/neu positivity (46%) compared to Chinese (25%), Japanese (14%) and Korean (29%) groups (P=0.002). Filipinos and Koreans had a higher proportion of ER negative cancers and Grade III cancers compared to Chinese and Japanese (P=0.001 for ER status and P=0.01 for grade). In our series, Korean subjects were significantly younger than the other three ethnic groups (P<0.0001 by AVOVA test).Conclusions: Differences in BC subtypes exist among Asian women with invasive breast cancer in New York. Filipino women are significantly more likely to have HER-2/neu positive BC compared with Chinese, Japanese, and Korean women. Our results are supportive of results from a study in which differences in distribution of Her2/neu positive cancers among Asians in California were seen. Furthermore, these findings provide one explanation for the worse clinical outcome for Filipinos compared with other Asians that has been previously reported.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3067.
Prognostic Value of Lymphogenic Micrometastasis of Patients with Breast Carcinoma: A Multicenter Cohort Study.
Authors: Gobardhan P. , Madsen E. , Elias S. , van Wely B. , van den Wildenberg F. , Theunissen E. , Ernst M. , van der Pol C. , Borel Rinkes I. , Wijsman J. , Vles W. , de Hooge P. , Ruitenberg H. , Perre C. , van Dalen T.
Published: December 2010

Introduction: Since the introduction of sentinel lymph node biopsy (SLNB) for staging breast cancer, lymphogenic micrometastases are commonly detected. The prognostic meaning of these small lymph node metastases and the consequences regarding the indication for adjuvant systemic treatment is under debate. Currently, robust data with a long time follow up are lacking.Method: Between January 2000 and December 2002 1411 patients with a cT1-2N0 breast carcinoma underwent surgery in seven hospitals in the Netherlands. Pathological examination of the sentinel node consisted of serial sectioning of SLN's and H&E and IHC staining. Based on the presence of lymph node metastases patients where divided into four groups: pN0 (n = 922), pN1micro (n = 103), pN1a (n = 285) and pN≥1b (n = 101). Median follow up was 77 months.Results: At the end of follow-up 184 (13.0%) patients had died. Breast cancer recurred in 244 (17.3%) patients, 165 of them having distant metastases (11.7%) patients. The 5- years overall survival was 93% for patients with pN0 disease, 94% for pN1micro, 88% for pN1a and 78% for pN≥1b (p < 0.001). The 5-years disease free survival was 87%, 90%, 85% and 72% (p < 0.001) respectively.Following adjustment for possible confounding characteristics and for adjuvant systemic treatment, overall survival was similar for pN0 en pN1micro patients and significantly worse for pN1a and pN≥1b (HR 1.18; 95% CI 0.58-2.39, HR 2.47; 95% CI 1.69-3.63, HR 4.36; 95% CI 2.70-7.04 respectively). Disease free survival was similar too in the pN0 and pN1micro group, and worse for pN1a and pN≥1b (HR 0.96; 95% CI 0.56-1.67 vs. HR 1.64; 95% CI 1.19-2.27, HR 2.95; CI 1.98-4.42). Distant metastases were more commonly observed in the pN1micro group than in the pN0 group (HR 1.22; 95% CI 0.60-2.49), but not significantly and far less than in the pN1a and pN≥1b group (HR 2.26; 95% CI 1.49-3.40, HR 3.49; 95% CI 2.12-5.77).Conclusion: After a relative long time of follow up disease free and overall survival for patients with micrometastasis in SLNs is comparable to patients without lymphogenic metastasis and more favourable than patients with macrometastasis. The presence of micrometastatic disease in the SLN is in itself not an indication for adjuvant systemic therapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 307.
Importance of Providing Tailored Resources to Patients with Metastatic Breast Cancer: Results of the Global BRIDGE Survey.
Authors: Mayer M. , Hunis A. , Oratz R. , Glennon C. , Spicer P. , Caplan E. , Fallowfield L.
Published: December 2010

IntroductionLife expectancy for women with metastatic breast cancer (MBC) has increased as advances have been made in treatment and management. Consequently, when measuring the burden of MBC, womens' experiences, quality of life, and gaps in information and support needs should be considered. The BRIDGE Survey (Bridging Gaps, Expanding Outreach – Metastatic Breast Cancer Patient Survey) was conducted in the US, Europe, Latin America and Africa to evaluate patients' (pts) own perceptions of living with MBC; results are presented here.MethodsA Steering Committee of 3 international oncology thought leaders, 4 pt advocates, and a global public health organization was set up with the support of Pfizer Inc. to develop a quantitative survey (Harris Interactive®). To provide a balance between low and high-resource countries, women in the US, UK, France, Spain, Poland, Belgium, Argentina, Mexico, and Egypt aged ≥18 years with stage IV/metastatic disease were recruited and interviewed. Survey methods included telephone, mail, and in-person interviews, dependent on country and pt preference. The 34-item questionnaire took ∼25-mins to complete. Topics included MBC resources, support networks, attention given to MBC vs. early breast cancer (EBC) and personal impact of the disease. Pts were also asked if they had ever participated in, or searched for information about, clinical trials.ResultsIn total, 950 women completed the survey, referred by ∼20 physicians in each of the 9 participating countries. Most women (53%) felt that MBC was given too little consideration compared with EBC. 75% actively sought information on MBC but 51% felt that available information did not address their needs. Written materials were most frequently used, although many women found the internet most helpful. Those women who sought information were more likely to take an active role in treatment decisions and be more optimistic in outlook. Globally, 18% of women reported that they had participated in a clinical trial, with encouragement to do so by a healthcare professional being the most common reason given (77%). Numerous country-specific differences were observed, most notably in ease of finding information, whether available information meets pt needs, need for medical vs. non-medical information, and clinical trial participation. There were also country-specific differences in womens' level of optimism on diagnosis.ConclusionsThese results suggest that women with MBC have different needs from those of pts with EBC and that these needs are not always being met. Women are asking for information relevant both to their culture and to their disease stage. Further results will be presented, and will be discussed in the context of a recently published MBC Consensus Report compiled by the MBC Advocacy Working Group (Community Oncology 2008;5[12]; Breast 2009 [In Press]).

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3085.
EGFR Gene Copy Number and Response to Lapatinib/Capecitabine in HER2 Positive Metastatic Breast Cancer Patients Pretreated with Trastuzumab.
Authors: Fabi A. , Graziano V. , Merola R. , Metro G. , Papaldo P. , Cianciulli A. , Mottolese M. , Sperduti I. , Vico E. , Cognetti F.
Published: December 2010

Background: Lapatinib (L) is a potent and selective dual inhibitor of EGFR and HER2. In HER2 positive metastatic breast cancer (MBC) pretreated with trastuzumab-based therapies, the association of L + Capecitabine (C) showed superior response rate (RR) and progression free survival (PFS) over C alone. We analyzed the correlation between EGFR gene copy number and clinical outcome in a population undergoing treatment with L 1250 mg daily plus C 2000 mg/m2 days 1-14 every 21. Materials and methods: EGFR gene copy number was assessed by FISH analysis in paraffin-embedded tissue blocks from 30 HER2 positive patients who had failed at least one trastuzumab-based therapy in the metastatic setting. EGFR gene status was scored as: the mean value of EGFR gene copy number (GCN) per nucleus, the mean ratio between EGFR and Chromosome 7 and the percentage of cells displaying more than 5 EGFR GCN. A receiver operating characteristic (ROC) analysis was set up to define the cut-off of mean EGFR GCN. A value greater than 3.36 was able to discriminate responders versus non-responders. The primary end point of the study was identification of the EGFR FISH score that best predicts RR Results: Twenty-five patients were evaluable for EGFR FISH analysis. No sample displayed a homogeneous amplification of the EGFR gene by ratio (EGFR gene/CEP 7 ≥2). Patients characteristics were as follows: median age 52.4 (range 32-70), visceral disease in 20 patients (80%), brain metastases in 11 patients; median number of previous trastuzumab-based therapies was 2 (1-4). Twelve patients achieved a partial response for a RR of 48% (95% CI: 28.8-64.5). At a median follow up of, PFS was 7 months (95%CI: 5-8). Median overall survival has not been reached at the time of analysis. Mean EGFR GCN was > 3.36 in 10/25 patients. These patients showed a significantly higher RR (n=8/10; 80%) as compared to those with mean EGFR GCN ≤3.36 (n= 4/15; 27%), (p=0.009). No significant difference in terms of PFS was seen. Conclusion: high EGFR GCN might predict for an increased likelihood of responding to L. This is the first report associating increased EGFR GCN with sensitivity to L. These results warrant further investigation on a larger sample of patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3095.
Curcumin Modulates Tamoxifen Response in Resistant Breast Cancer Cells.
Authors: De Gasperi M. , Cavazos D. , deGraffenried L.
Published: December 2010

Despite significant improvement in both detection and treatment, breast cancer remains the second leading cause of cancer deaths for women in the United States. Tamoxifen is the most commonly used treatment for estrogen receptor positive (ER+) breast carcinoma, yet up to 50% of patients with metastatic disease present with de novo resistance to tamoxifen, and almost all patients will eventually become resistant to tamoxifen treatment. The mechanism(s) for this resistance appear to be largely dependent upon activation of growth factor signaling pathways. Aberrant activation of the PI3K/Akt and MAPK pathways and their cross talk with both the genomic and nongenomic activity of the ER is implicated in tamoxifen resistance. Breast cancer cell lines with constitutively activated Akt are characterized by high levels of the pro-survival transcription factor Nuclear Factor kappa B (NF-kB), estrogen-independent growth, and resistance to tamoxifen treatment. Studies have shown that curcumin, a polyphenol derived from the rhizome of the perennial herb Curcuma longa, is able to inhibit the activation of NF-kB, which is implicated as one mechanism by which breast cancer cells become resistant to tamoxifen treatment. Previous studies in our laboratory have demonstrated that inhibition of NF-kB is sufficient to sensitize MCF-7 cells that have constitutively activated Akt and elevated NF-kB levels to tamoxifen treatment. However, clinical NF-kB inhibitors such as Velcade generally involve suppression of the proteosome, leading to severe toxicities. Therefore, identification of alternative approaches for suppressing NF-kB activity in the absence of toxicity could prove beneficial for enhancing response to tamoxifen. In the present study, we assessed the efficacy of curcumin at inhibiting breast cancer cell proliferation and survival both as a single agent and in combination with tamoxifen in MCF-7 cells that expressed both “normal” levels of Akt (Control) activity as well as those that expressed the constitutively active form of Akt (myrAkt). As has been previously reported, curcumin treatment inhibited survival and proliferation in the Control MCF-7 cells, but importantly curcumin was also able to suppress survival and proliferation in the myrAKT MCF-7 cells. Curcumin and tamoxifen co-treatment was found to synergistically inhibit survival in myrAKT MCF-7 cells. These findings demonstrate that curcumin can sensitize myrAKT MCF-7 cells to tamoxifen treatment. This preliminary data suggests that the combination of natural NF-kB inhibitors such as curcumin with tamoxifen may be a viable strategy to either prevent tamoxifen resistant disease or to re-sensitize refractory disease to tamoxifen treatment.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3098.
Evaluation of Costs and Impact in Quality of Life in Patients with Surgical Site Complications after Breast Cancer Surgery.
Authors: Arrieta-Joffe P. , Cerezo O. , Bargallo E. , Cruz E. , Lozano A. , Vilar-Compte D.
Published: December 2010

Background: Surgical site complications (infection, dehiscence, necrosis) continue to be an important cause of morbidity in patients undergoing breast cancer surgery. It seems to have a negative impact in quality of life and an increase in health-care costs in comparison with patients with a satisfying outcome.Meterial and Methods:Using two questionnaires to evaluate quality of life from the European Organization for the Research and Treatment of Cancer (EORTC), QLQ-C30 and BR-23, we analyzed 90 breast cancer patients after surgery. We obtained information about direct costs (gloves, gauze, antibiotics, wound dressing, hospital admission) and indirect costs (travel time, transport, day off at work) related to post-surgery consults. We analyzed dissimilarities concerning self-perception of quality of life between patients with and without surgical complications and differences in health-care treatment costs.Results: Twelve (11.76%) patients developed surgical site infection (SSI), 6 (5.88%) necrosis and 9 (8.8%) dehiscence.Self-perception of quality of life was decreased in patients with surgical site complications.The crude median costs of complicated patients was $1,368 US dollars compared to $992 US dollars in non-complicated patients. After adjusting costs according to type of complications only statistical significance was found in patients with SSI and dehiscence (95% confidence interval, $203-$301 and $94-$170 respectively) and the direct costs showed the most important difference.Discussion: The direct and indirect costs was superior in complicated than in non-complicated patients and there was a decrease in self-percieved quality of life, which is consistant with other findings. This should awaken interest in applying new methods to reduce risk of surgical site complications. There is an important lack of evidence regarding quality of life and costs related to post-surgery complications in any health care facilities in our country, which wil make this the first study to do it, specially in breast cancer patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3110.
Lymph Node Transplantation in Breast Reconstruction Using Perforator Flaps.
Authors: LoTempio M. , Studinger R. , Vaisille J. , Chen C. , Levine J. , Allen R.
Published: December 2010

Introduction: For many women undergoing breast reconstruction, the negative impact of upper extremity lymphedema outweighs the benefits of breast reconstruction.Patients with lymphedema present a challenging problem, and typically is not addressed during the reconstruction process. Lymph node dissections have decreased secondary due to the advent of the sentinel node biopsy. However, 7% of patients undergoing sentinel lymph node biopsy develop lymphedema. Previous treatments options have not proved to be highly successful, and include lymphatic massage and lymphaovenous anastomosis. We introduce a combined treatment for breast reconstruction using deep inferior epigastric perforator flap (DIEP) along with lymph node transplantation for lymphedema.Methods: Beginning in 2007, 23 patients have undergone bilateral breast reconstruction using the DIEP along with simultaneous lymph node transplantation. Each patient had demonstrated upper extremity lymphedema by measurements and subjective findings. Of the 23 patients 18 had radiation therapy and five had sentinel node dissection. The lymph node flap was harvested surrounding the superficial circumflex vessels in conjunction with the DIEP. The combined DIEP and lymph nodes were anastomosed to the internal mammary artery. Preparation of the axilla included removal of scar tissue, and fixation of the lymph nodes into the axilla.Results: Each patient had an uncomplicated postoperative course. These patients started to experience resolutions of lymphedema as early as ten days after surgery and continued up to six months. Currently, 15 patients no longer need lymphatic massage nor wear compression garment. One patient had a recurrence of breast cancer. Each patient reported their arm circumference had improved and decreased morbidity associated with lymphedema.Conclusion: Lymph node transplantation is a relatively new and exciting option for the management of lymphedema. Women who undergo breast reconstruction using perforator flaps can have simultaneous lymph node transplantation with minimal morbidity. These patients experienced an overall 89% improvement of symptoms.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3112.