bluuebunny/crossref_metadata_2025_split huggingface dataset:
Critical role of FoxO3a transcription factor in response to therapy with EGFR inhibitors, gefitinib and lapatinib, in breast cancer
Authors: Krol J. , Lam E. , Coombes C. 
Published: March 2020

21018 Background: Inhibition of the EGF-receptor is a potential therapeutic strategy for breast cancer. The PI3-K/PKB signalling pathway plays an important role in cell proliferation, survival and malignant transformation. The Forkhead box group O transcriptional factor, FoxO3a, is a direct downstream target of PKB. Here we studied the role of the FoxO3a in response to the EGFR inhibitors Gefitinib and Lapatinib. Methods: Response of breast cancer cell lines to treatment with Gefitinib and Lapatinib was evaluated by proliferation assay and FACS analysis. Expression of FoxO3a and its downstream targets was analysed using Western blotting. Sub-cellular localisation of FoxO3a was analysed by confocal microscopy. FoxO3a mRNA level was measured by RTq-PCR. siRNA transfections were performed using Oligofectamine reagent. Immunohistological staining was performed on patient samples to validate our findings. Results: Treatment of a panel of breast cancer cell lines with both inhibitors resulted in decreased proliferation due to G1-arrest and apoptosis in two sensitive cell-lines, BT474 and SKBR3. Western blot analysis revealed that response to the treatment was associated with a decrease in PKB and FoxO3a phosphorylation and thus the nuclear relocalisation of FoxO3a. Confocal microscopy confirmed that treatment induced nuclear translocation of FoxO3a only in sensitive but not resistant cells. This relocation of FoxO3a was accompanied by an induction of FoxO3a mRNA and its principal targets, p27kip1 and Bim. Transfection of the sensitive BT474 cells with FoxO3a specific siRNA resulted in a significant reduction in FoxO3a protein expression and Gefitinib-induced cell death. Comparison of immunohistochemical staining of biopsies from breast cancer patients before and after Gefitinib treatment revealed that therapy resulted in a significant increase in nuclear FoxO3a staining. Conclusion: Our results demonstrate for the first time that the EGFR-family inhibitors, Gefitinib and Lapatinib, specifically target FoxO3a to induce cell cycle arrest and apoptosis. This finding helps to define the mechanism of action of Gefitinib and Lapatinib and may provide novel insights into the molecular basis for resistance to these inhibitors. No significant financial relationships to disclose.
Quantitative measurement of HER2 expression and HER2 homo-dimerization in formalin-fixed paraffin-embedded (FFPE) breast cancer tissues using a novel proximity-based assay
Authors: Huang W. , Tan Y. , Jin X. , Mukherjee A. , Sperinde J. , Pannu H. , Prasad U. , Penuel E. , Winslow J. 
Published: March 2020

21023 Background: The best method to assess HER2 status remains controversial. Here we describe a novel assay that provides reliable quantitation of total HER2 (H2T) and HER2 homodimer (H22D) in FFPE tissues. The assay was cross-validated with conventional HER2 tests - immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Characteristics of the new assay and its potential application as predictive test were explored. Methods: Electrophoretic Tag (eTag) is a proprietary proximity-based technology that measures gene and protein expression. We have developed a novel application of the technology to quantify H2T and H22D in FFPE tissues. The validity of the assay was evaluated by analyzing its performance in cell lines and FFPE breast cancers. IHC (clone CB-11, Ventana) was performed in 174 breast cancers, scored according to standard IHC criteria and Histoscore (H-score), and HER2 gene amplification was evaluated by FISH (PathVysion, Vysis) in 19 breast cancers. eTag assays were performed on the same specimens and the results were compared with those of IHC and FISH. Results: H2T and H22D were proportional to the known expression levels in cell lines. H2T demonstrated a continuous measurement over a wide dynamic range (>2 logs), in contrast with conventional IHC (0, 1+, 2+, 3+). The correlation between H2T and IHC categories was significant (correlation=0.84, p<0.0001 by the Jonckheere-Terpstra test for trend), but overlap of H2T values in adjacent IHC categories was observed. H-score showed good correlation within the lower range of H2T, but plateaued at higher range of H2T. HER2 gene amplification (HER2/CEP17 ratio and gene copy #) by FISH was generally correlated with IHC categories and H2T values. There was a good correlation between H22D and H2T (r2= 0.7, P < 0.001). Conclusions: eTag reliably measures H2T and H22D in FFPE tissues. The continuous measure of H2T over a wide dynamic range and the novel H22D measure provide data superior to conventional HER2 tests by IHC and FISH, but clinical utility of the eTag assay remains to be investigated. eTag has potential as a predictive test for targeted HER2 therapy in breast cancer. No significant financial relationships to disclose.
Overexpression of claudin-11 decreases the invasive potential of bladder cancer cells in vitro, but has no effect on their electrical resistance
Authors: Awsare N. S. , Martin T. A. , Kynaston H. G. , Mansel R. E. , Matthews P. N. , Jiang W. G. ,   Metastasis & Angiogenesis Research Group
Published: March 2020

21038 Background: An increasing body of evidence suggests that tight junctions (TJ) play an important role in invasion and metastasis. Claudins (Cl), a group of integral membrane proteins play an important role in maintaining TJ structure and function. Hepatocyte growth factor (HGF) has been shown to disrupt TJ structure. We investigated the effect of forced overexpression of Cl-11 on TJ function and invasive potential of T24 bladder cancer cells, in response to HGF. Methods: Expression of Cl-11 in bladder cancer cells was determined by RT-PCR. Cl-11 expression construct was made from full length human Cl-11 obtained from normal human cDNA bank, using pcDNA 3.1/ NT-GFP-TOPO expression vector. T24 cells were transfected with the resultant expression cassette (T24Cl-11) or an empty plasmid control (T24GFP). Change in TJ function of stably transfected cells was assessed by trans-urothelial resistance (TUR). In- vitro matrigel adhesion and invasion assays were used to determine the malignant potential of the cells. Cells were treated with HGF at 50 ng/ml for TUR and invasion assay. Immunofluorescence was used to localise Cl-11 within the cell. Results: Successful transfection of T24 cells was confirmed by endogenous GFP fluorescence and by an enhanced Cl-11 mRNA signal in T24Cl-11 compared to wild- type (T24WT) or T24GFP. There was no difference in TUR between the three cell types. Adhesion was significantly higher in T24Cl-11 than in T24GFP [median (IQR); 0.11 (0.096–0.137) vs 0.095 (0.089- 0.115), p=0.045]. T24Cl-11 cells had significantly reduced invasiveness in response to HGF, compared to T24GFP [mean (95% CI); 2.4 (1.8–3.01) vs 3.31 (2.89–3.71), p=0.032]. Immunoflourescence staining with Cl-11 showed redistribution of Cl-11 protein to the cytoplasm and perinuclear regions. Conclusion: Cl-11does not appear to be involved in the TJ function in T24 bladder cancer cells; however, its cytoplasmic location and its effect on the metastatic potential of these cells suggests that it may have a role in modulating the cytoskeletal or signalling network of the cell. Further studies are required to elucidate the exact mechanisms involved. No significant financial relationships to disclose.
Clinical significance of proliferative activity evaluated by MIB-1 in the treatment and postoperative follow-up of early breast cancer
Authors: Nishimura R. , Arima N. 
Published: March 2020

21054 Background: To evaluate a clinical significance of proliferative activity in breast cancer, we studied relationships between proliferative activity and recurrence rate, the time of recurrence or adjuvant therapy. Methods: We analyzed 2448 patients with primary breast cancer between 1987 and 2004 in the Kumamoto City Hospital, and 437 cases out of the patients developed recurrence. Furthermore, the rate of recurrence before and after 1999 when postoperative adjuvant therapy (such as CEF or Taxanes) was started as standard treatment was investigated. Proliferative activity was judged by immunostaining for MIB-1. The fraction of proliferating cells was classified into 3 degrees (=19%, 20–49%, 50%=). Median observation period was 70 months. Results: 1) Distribution of patients by proliferation was as follows; =19%:1215 cases(50%), 20–49%: 870 cases(35%), or 50%=: 363 cases(15%). There was a significant relationship between proliferative activity and tumor size, nodal status, ER, PgR, p53 or HER2 status. 2) Multivariate analysis for disease-free survival revealed that a proliferative activity was one of significant factors in node-negative and positive cases. Recurrence rate was 11.6% in cases with low proliferation and 31.0% in high proliferation. The mean period from operation to recurrence in cases with low proliferation was 50.2 months, whereas 19.9 months in high proliferation (p<0.0001). Moreover, 74% of recurrent cases with high proliferation recurred within 2 years after operation, and there were few recurrences from the fifth year. 3) Patients with low proliferation frequently developed bone metastasis. In local recurrence, diffuse skin recurrence was often seen in cases with high proliferation. 4) The prognosis of patients in the later period (standard therapy group) was significantly better than that of patients in the earlier period, especially in high proliferation group. Conclusions: Proliferative activity might reflect aggressive behavior of breast cancer and predict the time of recurrence. The standard adjuvant therapy was effective in inhibiting early recurrence with high proliferation. It is important to take proliferative activity into consideration in the treatment and follow-up of breast cancer. No significant financial relationships to disclose.
Relevance of circulating tumor cells (CTC) in the progression of metastatic breast cancer
Authors: Altimari A. , Quercia S. , Benedettini E. , Rosati M. , Capizzi E. , Bernardi A. , Fiorentino M. , Zamagni C. , D'Errico Grigioni A. , Martoni A. A. 
Published: March 2020

21066 Background: metastatic breast cancer is an incurable disease. Molecular tumor staging methods are required for a better therapeutic approach. Patients and Methods: eight mL blood samples were drawn from 20 patients (pts) (mean age 61.9±9) with metastatic breast cancer at the time of the beginning of a new chemo- or hormonal treatment line. Median follow-up was 7±4.5 months. Twenty- five female healthy blood donor volunteers were used as controls. Blood samples were screened for the presence of circulating tumor cells (CTC) with a quantitative PCR method using molecular probes for human mammaglobin (hMAM) and cytokeratin 19 (CK19). Relative quantification of hMAM and CK19 RNA was obtained using a blood sample infected with known concentrations of MDA-MB453 cells as calibrator. Results: none of the healthy controls was hMAM positive while the cut-off for CK19 was calculated as the mean value of the controls plus two standard deviations. Nine of 20 (45%) pts were hMAM positive, 15/20 (75%) CK19 positive, while 18/20 (90%) pts were positive for at least one marker (CTC positive). Among the 14 (70%) pts who had a disease progression 7 died. Seven of 9 hMAM positive (78%) and 11/15 (73%) CK19 positive pts had disease progression. All the 14 pts who had disease progression were CTC positive while the only 2 CTC negative pts were free from progression at the time of last follow-up. Conclusions: (1) Multiple (hMAM, CK19) rather than single marker detection should be preferred for CTC blood screening in breast cancer. (2) Combined hMAM and CK19 evaluation might identify pts at higher risk of tumor progression and might be useful for stratification and decision making in metastatic breast cancer pts. No significant financial relationships to disclose.
Comparison between epidermal growth factor receptor (EGFR) gene in primary non-small cell lung cancer (NSCLC) and in fine needle aspirates from metastatic sites
Authors: Bozzetti C. , Tiseo M. , Lagrasta C. , Nizzoli R. , Leonardi F. , Gasparro D. , Spiritelli E. , Franciosi V. , Rindi G. , Ardizzoni A. 
Published: March 2020

21081 Background: Epidermal growth factor receptor (EGFR) gene copy number obtained by fluorescence in situ hybridization (FISH) has been recently proposed to predict which non-small cell lung cancer patients (NSCLC) are expected to benefit from EGFR tyrosine kinase inhibitors. However, it is still unknown whether EGFR status differs in metastases compared to primary NSCLC. In all studies, FISH have been performed on histological material. The possibility to perform FISH analysis on cytological material obtained by fine-needle aspiration from superficial and deep metastases would allow to know the real EGFR status when the metastatic site is not accessible for biopsy. Methods: EGFR gene copy number was analyzed by FISH on fine-needle aspirates (FNAs) obtained from 18 patients with metastatic NSCLC (11 lymphnod, 2 liver, 1 abdomen, 2 skin, 2 pleural effusion) and results were compared with those obtained on corresponding paraffin histological sections from the primary tumor. Results: The feasibility of EGFR-FISH on cytology was 89%; 2/18 samples were not evaluable because of lack of hybridization. EGFR-FISH was positive (= 4 EGFR copy number; Cappuzzo F. et al, JNCI 2005) in 69% (11/16) of the metastases and in 38% (6/16) of the primary tumors. Five of the 16 cases were EGFR positive on both primary tumor and metastatic site and 4 were negative on both primary tumor and metastasis. Seven cases (44%) showed primary tumor vs. metastasis discordance: in 6 cases EGFR was positive in the metastatic site but not in the primary tumor, while in one sample EGFR was positive in the primary tumor but not in the metastasis. Conclusions: EGFR-FISH can be reliably assessed on FNAs obtained from NSCLC metastases. Possible changes in EGFR status during the metastatic progression as well as cancer heterogeneity may account for discrepancies observed between primary tumor and metastatic sites. These findings should be considered in future studies which will be design to better elucidate the predictive role of EGFR-FISH in NSCLC. No significant financial relationships to disclose.
Relationship between BRCA1 promoter methylation and sensitivity of breast cancer cell lines to cisplatin and paclitaxel
Authors: Collins C. , Huo D. , Xu J. , Bleibel W. K. , Dolan M. E. , Olopade O. I. , Nanda R. 
Published: March 2020

21083 Background: While BRCA1 germline mutations are uncommon, and contribute to fewer than 5% of breast cancer cases, epigenetic alterations in BRCA1 occur more frequently. BRCA1 promoter methylation has been detected in 10–30% of breast tumors. Given the role of BRCA1 in DNA repair and cell cycle regulation, we hypothesize that cells with decreased expression of BRCA1 secondary to promoter methylation will be sensitive to DNA damaging agents and resistant to microtubule inhibitors, as has previously been shown for cells deficient in BRCA1 secondary to mutation. Methods: BRCA1 methylation was determined using methylation specific PCR (MSP) as previously described (Wei et al, Cancer Research 2005). The relative sensitivities of BRCA1 methylated, mutated and competent cells to cisplatin and paclitaxel were determined in five representative breast cancer cell lines using the AlamarBlue cytotoxicity assay. Exponentially growing cells were treated with increasing concentrations of cisplatin and paclitaxel for 96 hours. IC50 values and 95% confidence intervals (CI) were calculated from sigmoidal dose response curves fitted with SAS 9.1 Proc NLIN. Western blot analysis for BRCA1 was performed on each cell line. Results: Conclusions: Only one of the two BRCA1 methylated cell lines studied (UACC3199) was sensitive to cisplatin and resistant to paclitaxel, as hypothesized. While both cell lines are methylated, western blot analysis revealed that both express BRCA1, but to a lesser degree than unmethylated cells. BRCA1 methylation, as assessed by non-quantitative MSP, does not correlate with sensitivity to cisplatin and resistance to paclitaxel. Quantification of BRCA1 promoter methylation may better predict chemosensitivity. Identification of the degree of BRCA1 methylation which does correlates with sensitivity to cisplatin and resistance to paclitaxel could improve treatment selection for patients with breast cancer. This work was supported by the US Army Grant W81XWH-04–1-0545. [Table: see text] No significant financial relationships to disclose.
Expression of periostin in human breast cancer: Biological and clinico-pathological correlations
Authors: Puglisi F. , Minisini A. , Andreetta C. , Russo S. , Pegolo E. , Pascoletti G. , Mansutti M. , Fasola G. , Damante G. , Di Loreto C. 
Published: March 2020

21090 Background: Mesenchyme-specific genes are highly expressed by various types of human cancers. Recent lines of evidence suggest that periostin, a myoepithelial/basal gene, may play a role in the biology of breast cancer. Periostin is involved in metastatic process by taking part in mechanisms of adhesion and migration of epithelial cells. Furthermore, periostin seems to have a role in tumor angiogenesis through the up-regulation of vascular endothelial growth factor receptors in endothelial cells. We aimed at investigating the immunohistochemical expression of periostin in breast cancer and its correlation with established biological and prognostic factors. Methods: We performed immunohistochemical analysis of periostin in a consecutive series of 206 tumor samples from 200 patients with early breast carcinoma. To this aim, we used a specific antiperiostin antibody (Biovendor Laboratory Medicine, Inc., Palackeho 56, 612 00 Brno, Czech Republic). The intensity of immunoreactivity was scored as 0, 1, 2 or 3 denoting negative, weak, moderate and strong staining, respectively. Therefore, for statistical analysis, the periostin expression level for each case was putatively defined as positive if the predominant intensity was =1. We also analyzed immunohistochemical expression of Ki-67 (MIB-1), HER-2/neu, VEGF, VEGFR-1 and VEGFR- 2. Results: Periostin was found to be highly expressed by carcinoma cells, whereas it was absent in normal breast tissues. The pattern of expression was mainly cytoplasmic, although in some cases (11%) a nuclear reactivity was observed. A cytoplasmic expression was found in 108 out of 189 (57%) evaluable cases. Interestingly, a significant correlation was found between periostin expression and VEGF and VEGFR-1 (Spearman's rank test rho =0.37, p <0.0001 and rho=0.26, p=0.0005, respectively). Conclusions: These results strengthen the hypothesis of a relationship between periostin and vascular endothelial growth factor system. The role of periostin in angiogenesis and in other relevant molecular pathways in breast carcinoma deserves to be further investigated. No significant financial relationships to disclose.
Cancer incidence and mortality in the Caribbean
Authors: Phillips A. A. , Jacobson J. S. , Magai C. , Consedine N. , Horowicz-Mehler N. C. , Neugut A. I. 
Published: March 2020

21110 Background: Nearly 10% of immigrants to the United States come from the Caribbean region. In this paper, we analyzed incidence and mortality rates of the major cancers in the Bahamas, Barbados, Cuba, the Dominican Republic, Haiti, Jamaica, Puerto Rico, and Trinidad and Tobago, and compared them with US patterns. Methods: We obtained age-standardized, sex-specific cancer incidence and mortality rates for cancers of the bladder, breast, cervix, esophagus, large bowel, liver, lung, pancreas, prostate, and stomach for eight Caribbean countries and the US from the GLOBOCAN program of the International Agency for Research in Cancer (IARC) and for the US population from the Surveillance, Epidemiology, and End Results (SEER) Program of the NCI. Results: GLOBOCAN incidence and mortality rates for the overall US were lower than but correlated with overall SEER rates. Based on GLOBOCAN data, the incidence and mortality rates of cancers of the breast, prostate, large bowel, and lung, and, among males, bladder cancer were lower in the Caribbean countries than the US. Caribbean countries had higher rates of cancers of the cervix, esophagus, liver, and stomach. Haiti had the highest incidence and mortality rates of cervix and liver cancers. Jamaica and Haiti had the highest rates of stomach cancer. Conclusions: Cancer incidence and mortality in the Caribbean generally follow known patterns of association with economic development, infectious agents, and racial/ethnic origin. Studying these patterns and how immigration changes them may yield clues to cancer etiology. A better understanding of cancer incidence and mortality rates may help health policymakers to implement state-of-the-art treatment and preventive services for people of Caribbean descent both in their native countries and in immigrant communities in the US. [Table: see text] No significant financial relationships to disclose.
Receptor for advanced glycation end-products (RAGE) - soluble form (sRAGE) and gene polymorphisms in patients with breast cancer
Authors: Tesarova P. , Kalousova M. , Jachymova M. , Mestek O. , Petruzelka L. , Zima T. 
Published: March 2020

21113 Background: Receptor for advanced glycation end products (RAGE) may be involved in the pathogenesis of the cancer progression and metastasis. Pathological effects mediated via RAGE are physiologically inhibited by soluble RAGE (sRAGE), so the higher sRAGE levels may confer the patients with cancer with better outcome.. Our aim was to study sRAGE and RAGE gene polymorphisms in patients with breast cancer. Methods: We studied sRAGE and RAGE polymorphisms in 120 patients with breast cancer (subdivided based on the clinical stage, histologic grading, expression of hormonal and C-erb B2 receptors) and in 92 healthy controls. Results: Despite higher serum concentrations of AGEs, serum concentrations of sRAGE were lower in patients with breast cancer compared to healthy controls (1581 ± 777 vs. 1803 ± 632 ng/ml, p < 0.05). Serum levels of sRAGE were higher in patients with advanced breast cancer (stage III), lower grade and positive estrogen receptors and intermediate positivity of C-erb B2 (Her-neu) receptors and were also influenced genetically (G82S and 2184 AG polymorphisms of the RAGE gene). Conclusions: Decreased sRAGE levels in patients with breast cancer may contribute to the progression of the disease. Patients with better outcome (with low grade and positive estrogen receptors) have higher sRAGE levels. Progression of the disease, may, however, increase sRAGE levels, possibly as a compensatory mechanism to counteract further progression. No significant financial relationships to disclose.