bluuebunny/crossref_metadata_2025_split huggingface dataset:
Plasma DNA: A molecular marker for the pattern of dissemination of esophageal cancer
Authors: Banki F. , Hagen J. A. , Mason R. J. , DeMeester S. R. , Oh D. , Yacoub W. N. , Danenberg P. V. , DeMeester T. R. 
Published: March 2020

21015 Background: We previously showed that free DNA is present in blood of normal subjects and that the plasma DNA level is higher in patients with esophageal cancer compared to normal subjects. These observations suggest that the plasma DNA level may be a useful molecular marker in the diagnosis of esophageal cancer. We hypothesize that lymphatic and hematogenous spread may result in different plasma DNA levels despite both representing advanced disease. Our aim was to measure and compare plasma DNA levels in normal subjects, patients with localized esophageal cancer, patients with lymph node metastases, and patients with hematogenous spread to solid organs (systemic metastases). Methods: Plasma DNA was measured using PCR in 44 normal subjects, 25 patients with localized esophageal cancer (T1–3, N0, M0), 18 patients with lymph node metastases, and 7 patients with systemic metastases. Results: The 95th percentile of plasma DNA level in normal subjects was 19 ng/ml. The median plasma DNA levels were higher in all 3 groups of patients with esophageal cancer compared to control subjects. While there was no difference between the plasma DNA level in patients with localized esophageal cancer and those with lymph node metastases (median of 5 nodes involved, range: 1–31), patients with systemic metastases had a significantly higher level of plasma DNA compared to patients with lymph node metastases and those with localized esophageal cancer. Conclusion: Plasma DNA levels are elevated in patients with esophageal cancer, whether localized or associated with lymph node metastases. In patients with systemic metastases, a significant additional increase in the plasma DNA level occurs. This suggests that measuring the plasma DNA level is a useful molecular diagnostic tool for detection of disseminated esophageal cancer. [Table: see text] No significant financial relationships to disclose.
The use of CA15.3 in breast cancer: A single institution experience
Authors: Sheikh R. , Behbehani A. T. , Peader M. , McCaffrey J. 
Published: March 2020

21017 Background: Breast cancer is a significant cause of morbidity and mortality worldwide. Recent advanced its treatment are promising. Gathering prognostic information may optimise treatment delivery. In addition to clinical parameters, the use of tumour markers may aid this process. CA 15.3, is a transmembrane glycoprotein.. Its clinical application varies from detecting disease recurrence in asymptomatic patients, assessing response in metastatic setting and monitoring the clinical course of the disease. Methods: A retrospective analysis was conducted evaluating medical record data of all the patients treated for breast cancer in our institution between 1999- 2005. CA15.3 was carried out using Roche 2010 (Elecsys system). Results: A total of 176 patients were identified. Table (1) Group A, one hundred patients remained disease free at follows up (58%). 95 patients had their CA15.3 within normal range, a diagnostic specificity of 95%. Group B, 61 patients developed metastatic/recurrent disease at follow up. 39 patients (64%) developed metastatic disease later. Group C, 15 patients had metastatic disease at diagnosis. We found that CA15.3 has a sensitivity value of 64% and a specificity of 95% in our institution.. This correlates well with ASCO review 67% and 92% respectively. In patients with no evidence of metastatic disease at diagnosis, a postive CA15.3 during follow up is strongly suggestive of disease recurrence. Giving a positive predictive value of 89%. In Two patients (5%) CA15.3 was the first predictor of recurrence. Radiology was the first indicator of recurrence in six cases (15%), while both radiological and serological correlation was noted in 31 patients (80%). Conclusions: We found CA15.3 a reliable marker in assessing response for therapy. In clinical practice, raised CA15.3 can be incorporated in decision making to change current chemotherapeutic regimen whenever there is a delay in obtaining prompt radiological evaluation. [Table: see text] No significant financial relationships to disclose.
Qualitative age interactions (or effect modification) suggest divergent pathways for early-onset and late-onset breast cancers
Authors: Anderson W. F. , Brinton L. A. , Chen B. , Devesa S. S. 
Published: March 2020

21036 Background: Notwithstanding some recent declines, breast cancer incidence rates have risen for decades, though not equally for all age groups. We used the National Cancer Institute's SEER program to further explore the effect of aging upon breast cancer incidence. Materials and Methods: The SEER program collected data on n=494,543 in-situ + invasive female breast cancer cases, newly diagnosed during 1974–2003. Temporal trends by race, stage, and grade were stratified by age at diagnosis in decades: 20–29 to 80+ years. Results: We observed age interactions over time. For example, as the specification of grade improved from 1974–2003, temporal trends for high and low grade tumors varied with age. Among women ages <40 years, high grade lesions were more common than low grade tumors for all time periods. Among women ages 40+ years, high grade lesions were more common during the early years, and then rates crossed, after which low-grade tumors were more common than high grade lesions. Conclusion: Age at diagnosis was both a quantitative (non- crossover) and qualitative (crossover) effect modifier. The crossing of rates from high to low grade tumors among women ages 40+ years in the 1980s is consistent with more aggressive breast cancer screening, with mammography preferentially detecting low grade tumors among women targeted for screening, i.e., ages 40–80 years. Though once thought to be rare or artifactual, qualitative interactions or effect modification suggest etiologic heterogeneity in an otherwise homogeneous disease process. Indeed, if true, qualitative age interactions imply divergent pathways for early-onset and late-onset breast cancers. No significant financial relationships to disclose.
Women with Her2 unamplified but chromosome 17 hyperploid metatstatic breast cancer (MBC) respond to traztuzumab
Authors: Brufsky A. , Chow S. , Kapoor A. , Bhargava R. , Surti U. , Rosenzweig M. 
Published: March 2020

21051 Background: Guidelines for the interpretation of Her2 testing by FISH in determining therapy for Her2 positive metatstatic breast cancer generally suggest that only women with Her2 to chromosome 17 (CEP 17) amplification ratios of greater than 2.0 will benefit from traztuzumab. However, hyperploidy of chromosome 17 may lead to increased Her2 copy number despite a normal HER2/CEP 17 ratio, which may also predict response. We sought to examine hyperploid frequency in a well defined Her2 positive MBC population. We also sought to determine the time to progression (TTP) on first chemotherapy for MBC in such women, as well as their overall survival (OS) with MBC. Methods: A tissue mircoarray (TMA) with 3-fold redundancy was constructed using 0.6mm cores from primary or metatstatic paraffin embedded tumor from 124 cases which were Her2 2+ or 3+ by IHC, or amplified by FISH, on pathology report. Four micron thick TMA sections were used for HER2 IHC and HER2 FISH analysis. HER2/CEP 17 ratios of 2.0 or more was considered as amplification. If the tumor was 2+ or 3+ by IHC, greater than or equal to 3 signals for chromosome 17 were seen in a majority of cell nuclei, and the HER2/CEP17 ratio was less than 2.0, the tumor was considered hyperploid Her2 FISH negative. Results: Tissue blocks from 52 tumors delivered clearly interpretable amplification for Her2 by FISH. An additional 10 tumors (8%) were hyperploid, IHC positive (2+ or 3+), and FISH negative. Median survival (Kaplan-Meier) was 39 months (95% CI, 21–62) for women with hyperploid FISH negative tumors and 41 months (95% CI, 25–53) for women with FISH positive tumors (p=0.63, NS, Wilcoxon). Median time to progression (Kaplan-Meier) on first therapy for metastatic disease was 14 months (95% CI, 8–20) for hyperploid FISH negative tumors and 15 months (95% CI, 12–18) for women with FISH positive tumors (p=0.41, NS, Wilcoxon). Conclusions: In this analysis, women with hyperploid Her2 FISH negative IHC positive tumors had similar TTP and OS as women with Her2 FISH positive tumors. These women, although Her2 FISH negative, appeared to respond well to traztuzumab. These provocative results should be repeated on larger data sets. No significant financial relationships to disclose.
A novel rapid genetic diagnosis for peritoneal recurrence in gastrointestinal cancer patients
Authors: Iinuma H. , Okinaga K. , Watanabe T. , Fukushima R. , Inaba T. , Mimura T. , Matsuda K. , Ohnaka S. , Hayashi T. 
Published: March 2020

21052 Background: The prediction of peritoneal recurrence in gastrointestinal cancer patients is extremely important in order to prevent an unfavorable prognosis. The aim of our study was to evaluate the usefulness of a novel rapid RNA amplification system with transcription-reverse transcription concerted reaction (TRC) to detect isolated tumor cells in peritoneal washes of gastrointestinal cancer patients as an intra-operative genetic predictor of peritoneal recurrence. To clarify the characteristic qualities of TRC, we compared the results with those of one-step real-time RT-PCR. Methods: A total of 325 cytology-negative gastrointestinal cancer patients (164 gastric cancer, 161 colorectal cancer) were included. This study was approved by the institutional review board of our hospital and all patients provided written informed consent. Peritoneal washes of Douglas cavity were collected from patients. One-half of each wash was used for cytopathology and the other half was used for molecular diagnosis. Total RNA was extracted from cells which were separated from washes, and TRC and one-step real-time RT-PCR of multiple marker genes (CEA, CK20 mRNA) were performed using a TRCRapid-160 and LightCycler, respectively. Results: The rapidity and simplicity of the TRC is superior to that of one-step real-time RT-PCR. The detection time of marker genes by TRC was only 20 minutes as compared to 70 minutes of real-time RT-PCR. The sensitivity and reproducibility of the TRC were equal to those of real-time RT-PCR. Molecular detection of CEA and/or CK20 transcripts of the peritoneal washes by TRC showed a significant correlation to the depth of invasion, venous invasion, lymph node invasion, lymph node metastasis, peritoneal metastasis and stage. The peritoneal recurrence- free and overall survival rates of CEA and/or CK20 transcript-positive patients were significantly shorter than those of marker gene-negative patients. CEA and/or CK20 transcripts in peritoneal washes were independent prognostic factors of peritoneal recurrence-free survival and overall survival. Conclusion: TRC may be more useful than real-time RT-PCR due to its rapidity and simplicity for the intra-operative genetic diagnosis of peritoneal recurrence in patients with gastric and colorectal cancer. No significant financial relationships to disclose.
Suppression of thymidine phosphorylase expression by promoter methylation
Authors: Blanquicett C. , Guarcello V. , Naguib F. , el Kouni M. 
Published: March 2020

21100 Background: Thymidine phosphorylase (TP), an important enzyme in the pyrimidine salvage pathway, is also responsible for the activation of capecitabine (CAPE), a 5-FU prodrug. TP expression has been associated with response to CAPE. The regulatory mechanisms of TP expression in human tumor cells remain unclear. Since promoter methylation of CpG sites is a known mechanism of reversible silencing of gene expression, we investigated whether TP promoter methylation is involved in the regulation of TP expression. Methods: TP mRNA levels were determined in 5 cancer cells (SKBR-3, 786-O, HT-29, MDA-231, DLD-1) by Real- Time Quantitative PCR (RT-Q-PCR).The degree of methylation in these cells was identified by examining methylated CpG sites with restriction PCR and bisulfite sequencing. To analyze the TP promoter region, two fragments were PCR amplified from genomic DNA. Minimal promoter activity was determined by Luciferase reporter assays, and DNA-protein interactions were evaluated by Electrophoretic Mobility Shift Assays (EMSA). Results: SKBR-3 cells showed high TP expression, while DLD-1 cells did not express TP, as confirmed by examination of TP mRNA, protein, and enzyme activity. 786-O, HT29, and MDA-231 cells demonstrated intermediate TP expression. Methylation in the promoter, intron 1 and exon1 regions of the TP gene in SKBR-3 was lacking, while in DLD-1 cells, extensive promoter methylation was observed. Treatment of DLD-1 and SKBR-3 cells with the methylation-inhibitor, 5-aza-2-dC, resulted in a concentration-dependent increase in TP mRNA as well as protein levels in DLD-1 but not SKBR-3 cells. Trichostatin-A (TSA), a histone deacetylase inhibitor, improved the 5-aza-2dC induced TP reactivation. Gel shift assays demonstrated that methylation of the TP promoter significantly inhibited transcription factor binding. Supershift Analyses suggest that the Sp1 transcription factor has a role in the regulation of TP expression and to a lesser degree, so does Sp3. Conclusions: These results establish methylation of site-specific CpG in the TP promoter to be a putative mechanism of down-regulated TP expression in cancer cells. Insight into the TP regulatory mechanisms may have implications in CAPE therapy optimization and in modulating prognosis of cancer patients treated with CAPE. No significant financial relationships to disclose.
Evaluation of clinical significance of basal-like subtype of breast cancer identified by immunohistochemistry
Authors: Cassol L. B. , Graudenz M. , Zelmanowicz A. , Cancela A. , Coral C. , Yser G. , Garicochea B. 
Published: March 2020

21107 Background: At present time, prognostic evaluation of initial breast cancer is mostly based on patients’ clinical and tumoral histological features. Recently, molecular subtypes of invasive breast cancer were recognized through DNA microarray profiling studies. This new classification can potentially improve the prognostic evaluation but this technology is still not widely available. The possibility of identifying the molecular subtypes through simpler and cheaper methods is promising. The identification of basal-like breast cancer (BLBC) is particularly important because it has a poor outcome and fewer treatment options available. Additionally, Epidermal Growth Factor Receptor (EGFR) expression seems to be more frequent in this subtype. Methods: A retrospective cohort of 112 consecutive patients with pathologic stage I or II primary breast carcinomas, treated in the same institution between 1995 and 2000, was studied. Histological and clinical features as well as clinical outcome and survival were reviewed. Immunohistochemical analysis was carried out in representative blocks of tumors with antibodies against Estrogen Receptor (ER), Human Epidermal Growth Receptor - type 2 (HER2), CK5/6 and EGFR. The primary endpoint was to determine the prevalence of BLBC (ER and HER2-negative and CK 5/6 and/or EGFR-positive) in this population. Results: 13 of 112 tumors (11,6 %) were BLBC. Their mean age was 49 years; 77 % were stage II and 100 % were invasive ductal carcinomas. There was no prognostic difference between BLBC and the subtypes luminal (ER-positive and HER2-negative); HER2-overexpressing (HER2-positive) and undetermined (four markers negative) relating to disease-free and overall survival. This is probably related to limited number of patients in this study and good prognosis of initial-stage patients. 11 of 112 cases (10%) were positive for EGFR. EGFR-positive tumors tended to be ER- negative (23 % vs 5 %; p=0,01). Conclusions: Our results suggest that it is possible to identify BLBC by immunohistochemical analysis of ER, HER2, CK 5/6 and EGFR. Besides, EGFR expression seems to be more frequent in ER-negative tumors, which suggest EGFR-targeted drugs may benefit this population. No significant financial relationships to disclose.
Employing a blood based gene expression signature to detect early stage breast cancer
Authors: Tobin D. , Lindahl T. , Hagen N. , Bårdsen K. , Jensen M. , Sharma P. , Lönneborg A. , Børresen-Dale A. , Aarøe2 J. , Sæbø S. , Skåne P. 
Published: March 2020

21117 Background: Existing methods to detect breast cancer (BC) in asymptomatic patients have limitations, and there is a need to develop more accurate and convenient methods. We recently demonstrated the potential use of gene expression profiling in peripheral blood cells (PBC) for early detection of BC (1) and repeated this with a larger study using the Agilent platform with an accuracy of 75± 7%. Objective: 2 studies are presented that investigate: i) whether effective normalization of experimental conditions can improve diagnostic accuracy, ii) whether a blood based signature developed for BC can discriminate other forms of cancer, and iii) whether an expression signature developed using stage 0 patients can be used to predict BC in stage I disease, and vice versa. Material and Methods: Study I enrolled 60 females with BC and 60 healthy females. Study II enrolled 20 females with early stage BC (10 stage 0 and 10 stage I), 20 healthy females, and 8 females with colon cancer. Gene expression analysis was conducted using the ABI HGSM v2.0 with 32,878 oligo probes. Expression data were analyzed by PLSR for model building and results validated using cross-validation and test set validation. Results: Effective normalization of the data led to improved diagnostic performance. The signature developed using 20 BC and 20 non-BC samples classified 7/8 colon cancer patients as non-BC. The signature developed using stage 0 vs non-BC detected cancer in stage I patients, and the signature developed for stage 1 detected cancer in stage 0 patients. Conclusion: A blood-based gene expression signature can be developed for early stage breast cancer, which is specific and able to distinguish between other forms of malignancy such as colon cancer. The gene expression pattern is systemically affected in early stage BC patients in which there is typically no direct contact of blood cells with cancer cells. References: 1. Sharma P et al. (2005) Breast Cancer Res. 7 (5): R 634–44 2. Aaroe J et al (2006), poster no:125, 97th AACR Annual Meeting, Washington DC, USA Some RT-PCR analyses were performed by Marion Hirt IMGM Laboratories, Martinsried, Germany. [Table: see text]
Breast cancer submicrometastasis detection in sentinel lymph node by molecular biology techniques: A new tumor staging method?
Authors: Perazzo F. , Denninghoff V V. , Avagnina A. , Paesani F. , Allende D. , García A. , Crimi G. , Abalo E. , Elsner B. 
Published: March 2020

21120 Background: The purpose of this study was to analyze by molecular biology techniques (MBT) the presence of RNAm of mammaglobin (MAG) A and B in breast sentinel lymph nodes (SLN) by RT-PCR, and to correlate these findings with breast tumor characteristics. Methods: SLN from 51 patients with breast tumors smaller than or equal to 3 cm, with no palpable axillary lymphadenopathies were included. Hematoxylin-eosin (HE), immunohistochemistry study (IHC) and MBT were performed. Patients’ staging was assessed according to Saint Gallen's Consensus. Twenty-five patients had clinical follow-up. Results: HE showed metastasis in 4/51 cases, 5/51 were detected with IHC, and one of them negative for HE. MBT allowed the detection of 18/51 positive SN, which included the 5 above-mentioned cases. The other lymph nodes herein studied (33/51) showed no metastases with MBT. Patients’ mean age was 58 years, tumor's mean size was 0.96 cm. The average follow-up was 15 months (range: 1–29 months). The patient with a recurrence at 23 months of clinical follow-up was found to be the only one with HE (+), IHC (+), MAG-A (+) and MAG-B (+). Conclusions: SN study with HE fails to identify all SN with metastases. Immunohistochemistry and MBT increase metastasis detection. Multiplex RT-PCR technique for A and B MAG proves to be specific and sensitive. No direct relationship was found between the risk factors analyzed and the expression of MAG. Therefore, MAG could be used independently as a prognostic factor. The presence of hidden metastasis plays a significant role mainly in the overall survival rate of pre- and postmenopausal patients. This study will be useful to formulate hypotheses. Further research, including a larger population and a longer follow-up period, will be required to confirm these hypotheses. Should our findings be confirmed in the future, molecular biology determinations could modify patients staging and treatment. No significant financial relationships to disclose.
Molecular analysis of hormone refractory prostate cancer biopsies supports the rationale of using mTOR inhibitors
Authors: Stoss O. , Zielinski D. , Czeloth K. , Middel P. , Henkel T. , Albers P. 
Published: March 2020

21126 Background: The aim of our group is to identify molecular signatures in samples of hormone refractory prostate cancer (HRPC) patients (pts) that lead to novel rationales for medical treatment. We have previously shown that transurethral resections (TUR) of the prostate in HRPC pts are useful specimens for gene expression profiling using microarrays (ASCO 2006). Aim of this study was to prove the feasibility of gene expression profiling on prostate biopsies and to develop a standardised tissue handling protocol in order to facilitate multicenter research. Methods: Biopsy material and corresponding TUR chips or classical specimens from 8 pts with HRPC, 13 pts with localized PCA, 6 pts with benign prostatic hyperplasia (BPH) and 11 pts without cancer or BPH were investigated and compared. The tumor type and content was evaluated by a pathologist. Tissues were preserved in liquid nitrogen or RNAlater. Different tissue lysis and RNA purification methods were compared by the quantity (NanoDrop measurement) and quality (Bioanalyser, Agilent) of isolated RNA. Gene expression profiling occurred on Affymetrix HG-FOCUS arrays. Results: Most reliable gene expression results were obtained by biopsy lysis in Trizol using the QIAshredder. A total of more than 1 μg RNA was isolated from one biopsy. RNA quality fulfilled pre-defined criteria such as a 28S/18S rRNA ratio of > 0.8, an area under the curve of > 10% and a RNA integrity number > 6.5. A comparison of HRPC and PCA samples clearly confirmed previous results of a deregulation of protein biosynthesis (translation initiation and elongation factors, ribosome biogenesis) and PI3K signalling pathway components. Conclusions: Gene expression profiling supports the induction of the PI3K-AKT-mTOR pathway in HRPC. A standardised protocol for gene expression profiling from prostate biopsy samples applicable for translational research programs within multicenter clinical trials is now available. As a part of a clinical phase II trial that aims to investigate survival benefits on HRPC pts treated with docetaxel ± RAD001, a translational research program is now set up in parallel to identify biomarkers for response prediction using microarray gene expression analysis from prostate biopsies. No significant financial relationships to disclose.