bluuebunny/crossref_metadata_2025_split huggingface dataset:
Molecular profiling of circulating tumor cells in the peripheral blood of patients with primary and metastatic breast cancer
Authors: Fersis N. , Deckwart V. , Leitz A. , Weber M. , Rom J. , Böcher O. , Albert W. H. , Bastert G. , Sohn C. , Kaul S. 
Published: March 2020

20033 Background: The purpose of this study was detection and expression profiling of circulating tumor cells (CTC) in breast cancer patients. Methods: Two separate probes of 5 mL peripheral EDTA-blood from patients with primary breast cancer (n=167) and metastatic disease (n=111) were used for immunomagnetic tumor cell selection. Targets for preanalytical enrichment were the antigens EpCAM and MUC-1. Separated cells were lysed and used for mRNA isolation and c-DNA synthesis. The breast carcinoma-associated transcripts EpCAM, MUC-1, HER-2, claudin7, cytokeratin 19, mammaglobin 1, prostate-specific ets factor (PSE) and survivin were amplified by three separate multiplex RT-PCR reactions. Amplicons were analysed by capillary electrophoresis with the Agilent Bioanalyzer 2100. Specificity of the RT-PCR was confirmed by examination of blood of healthy donors. Results: Sensitivity for every single transcript was adjusted to 2 tumor cells per 5 ml blood. Tumor-associated transcripts were detected in 31 of of 167 (18.5%) patients with primary breast cancer and in 46 of 111 (41%) patients with metastatic disease. The marker with the highest incidence in both groups was MUC-1, with a positivity rate of 81%. Tumor-associated transcripts were heterogenouosly expressed, however multiple markers were identified in more than 50% of the positive samples. Conclusion: Using a combination of preanalytical immunomagnetic tumor cell enrichment followed by a multigen RT-PCR approach we describe a sensitive detection system for breast carcinoma cells. In this study a panel of 8 genes overexpressed at high levels in metastatic breast cancer was selected for the identification of disseminated tumor cells in the peripheral blood of breast cancer patients. HER-2, survivin as a unique member of the inhibitor of apotosis protein family, as well as PSE identified in circulating breast cancer cells may serve as prognostic indicators of tumor progression and could represent valid targets for new individualized therapeutic interventions. No significant financial relationships to disclose.
Differential spatial expression and activation pattern of EGFR and HER-2 in human breast cancer
Authors: Fink-Retter A. , Gschwantler-Kaulich D. , Hudelist G. , Mueller R. , Kubista E. , Czerwenka K. , Singer C. 
Published: March 2020

20041 Background: Overexpression of HER family members is a well-established prognositc factor and identifies potential targets for antibody-based receptor blocking strategies. While several studies have analyzed the expression of HER-2 and other HER-family members in malignant tumors, considerably less is known about their expression and activation in non-involved breast tissue from breast cancer patients. Methods: We have therefore investigated the differential expression of EGFR, HER-2 and their tyrosinekinase activated forms (ptyr-1248 HER-2 and ptyr-845 EGFR) by using immunohistochemistry (IHC) in 64 tumor specimens containing malignant epithelium (a), non-malignant tissue located at the peritumoral margin (b), and univolved breast tissue obtained at a distance of at least more than 5 cm from the malignant tumor (c). Results: We found significantly higher HER-2 protein expression levels in malignant epithelium than in non-malignant epithelium (p = 0.000004, Fisher’s Exact Test). Stromal HER-2 was absent in any of the tissues investigated. Epithelial EGFR expression did not differ between the three tissue types, but stromal EGFR protein was significantly more common in peritumoral and non-involved distant tissues when compared to tumor tissues (p = 0.008, Fisher’s Exact Test). We only found one case of intratumoral epithelial ptyr-1248 HER-2 expression, and interestingley two cases in peritumoral and peripheral normal tissue. Furthermore, we did not observe ptyr-845 EGFR in any of the samples analyzed.We found a significant overall correlation between epithelial and stromal EGFR expression (r = 0.442; p < 0.0001; Spearman’s Rho). Furthermore, we found a significant overall correlation between stromal EGFR expression and normal tissue type (r = 0.170; p < 0.02; Spearman’s Rho), and an inverse correlation between epithelial HER-2 and normal tissue type (r = 0.492; p < 0.0001; Spearman’s Rho). Conclusion: We have found a differential expression pattern of EGFR, HER-2, and activated HER-2 that is dependent on the spatial relation to a malignant tumor. Our observation of decreased intratumoral EGFR expression and the absence of activated EGFR suggest that, in contrast to HER-2, EGFR inhibition might not be an ideal target for antibody therapy. No significant financial relationships to disclose.
Role of metalloproteinase-7 in oxaliplatin acquired resistance in colorectal cancer cell lines
Authors: Almendro V. , Maurel J. , Augé J. , Laus G. , Domingo-Domenech J. , Fernández E. , Gascon P. 
Published: March 2020

20042 Background: Mechanisms responsible for acquired resistance in colorectal cancer (CRC) tumors are not well understood. Anticancer drugs have been shown to enhance FASL expression by NF-kB induction. Additionally Metalloproteinase (MMP)-7 is over-expressed in CRC and has been shown to inhibit apoptosis by cleavage of FASL. We have previously shown in vivo that, sFASL increment was associated with acquired chemoresistance. Therefore we speculate that inhibition of MMP-7 or NF-kB can reverse chemoresistance in CRC cell lines. Methods: We generated an oxaliplatin-resistant cells (HT29R) from a p53 mutated (HT29) cell line. Both cell lines were cultured for 72h with different concentrations of oxaliplatin, BAY11–7085 (inhibitor of NF-kB activation), 0.01 mM of the MMP-7 inhibitor 1,10-Phenanthroline monohydrate (1,10-PM) and 100 ng/ml of DX2 monoclonal antibody. Different drug combinations were performed. Citotoxicity was determined by the MTS method, and cell cycle was analysed at 72h. Cell lines were characterized for MMP-7 expression (ELISA), NF-KB (Western-Blot), Fas expression (immunohistochemistry) and FasL expression (Western-Blot). Results: FAS was down-expressed in HT29R compared to HT29. The HT29R cells showed a IC50 for oxaliplatin 2-fold higher than normal cells. Treatment with 1,10-PM decrease MMP-7 levels (p < 0.005) compared with untreated cells. Additionally, inhibition of MMP-7, restore IC50 values after oxaliplatin treatment in HT29R without changes in NF-KB expression. This oxaliplatin-resistant cell line, presents also sensibility for BAY11–7085, without affecting MMP-7 levels. Finally the addition of oxaliplatin to the MMP-7 inhibitor, increase FAS-mediated apoptosis (induced by DX2 antibody), suggesting that FASL cleavage is responsable of sensitivity. Conclusions: Reversal of oxaliplatin chemo-resistance can be obtained either by MMP-7 or NF-kB inhibition. Both drugs induced sFASL decrement, by inhibiting cleavage or expression, respectively. No significant financial relationships to disclose.
Temozolomide, irradiation and hypoxia promote homing of hematopoietic progenitor cells towards gliomas
Authors: Wick W. , Tabatabai G. , Frank B. , Weller M. 
Published: March 2020

20044 Background: Temozolomide and irradiation are essential parts of the standard therapy and hypoxia is a critical aspect of the microenvironment of gliomas. IN the present study, we aimed at investigating the impact of these stimuli on the previously defined transforming growth factor beta (TGF-β)- and stromal cell-derived factor-1/CXC chemokine ligand 12 (SDF-1α/CXCL12)-dependent migration of adult hematopoietic stem and progenitor cells (HPC) towards glioma cells in vitro and the homing to experimental gliomas in vivo. Hyperthermia served as control. Methods and Results: Cerebral irradiation of nude mice at 21 days after intracerebral implantation of LNT-229 glioma induces tumor satellite formation and enhances the glioma tropism of HPC in vivo. Supernatants of temozolomide-treated, irradiated or hypoxic LNT-229 glioma cells promote HPC migration in vitro. Reporter assays reveal that the CXCL12 promoter activity is enhanced in LNT-229 glioma cells at 24 h after irradiation at 8 Gy or after exposure to 1% oxygen for 12 h. The irradiation- and hypoxia-induced release of CXCL12 depends on hypoxia inducible factor-1 alpha (HIF-1α), but not on p53. Induction of transcriptional activity of HIF-1α by hypoxia and irradiation requires an intact signaling cascade of TGF-β. Conclusions: Thus, we delineate a novel stress signaling cascade in glioma cells involving TGF-β, HIF-1α and CXCL12. Stress stimuli can be temozolomide, irradiation and hypoxia but not hyperthermia. These data suggest that the use of HPC as cellular vectors in the treatment of glioblastoma may well be combined with anti-angiogenic therapies which induce tumor hypoxia. [Table: see text]
Hypoxia effect on non-malignant prostatic urethra during prostatectomy
Authors: Josephson J. W. , Vira M. , Gillespie J. W. , Pinto P. A. , Srinivasan R. , Chuaqui R. F. , Emmert-Buck M. R. , Erickson H. S. , Coleman J. A. 
Published: March 2020

20048 Background: With changes in technology and surgical technique paving the way to improved prostectomies, attention must be paid to the effects of hypoxia during variable surgical times. Hypoxia has been reported to play various roles that are implicated in the development of spurious molecular mutagenesis and tumorogenesis. However, the specifics leading to alterations and downstream effects in non-malignant tissues are unknown. Further, the consequence of hypoxia on tissues for analysis of tumor markers may be confounded. The objective of our study is to evaluate the effects of iatrogenic ischemia on surrounding non-malignant tissue in patients undergoing prostatectomy. As potential tissue markers to assess putative hypoxia tissue damage, gene expression levels in microdissected cell populations were evaluated. Methods: Normal prostatic urethra tissue was procured from five patients with laparoscopic 5 mm biopsy forceps at three time points during surgery and was immediately imbedded in OCT and placed on dry ice. First biopsy was taken prior to loss of blood supply after anterior bladder neck dissection. Second biopsy was taken directly after prostate was isolated from the patient. Third biopsy was taken ex vivo after vesico-urethral anastomosis. Urethral epithelium samples were normalized using the gold standard cell counting (2,000 microdissected cells) and quantitation of total RNA was conducted using NanoDrop technique. Gene expression analysis was assessed using quantitative RT-PCR (qRT-PCR) of four historically high and low abundant housekeeping genes (18s, ACTB, CYPA, and TFRC). Results: Housekeeping genes were up-regulated with an average four fold increase in transcript RNA at each time point respectively. Conclusion: These data suggest that hypoxia plays a putative significant role in the expression of genes in non-malignant tissues. The additional gene sets for analysis include traditional hypoxia related genes (e.g. HIF-1a, VEGF, AKT, and VHL) and non-hypoxia related genes (e.g. LAMP2, MTR, and STAT5B) to further investigate the prospective global up-regulation of gene expression. The conclusions may elucidate the role of hypoxia in disease processes and potentially its role in confounding or influencing tumor marker gene expression between patient samples. [Table: see text]
Circulating levels of the breast cancer growth factor GP88 in the serum of breast cancer (BC) patients
Authors: Serrero G. , Tkaczuk K. , Tait N. , Golubeva O. , Dai H. , Feldman F. S. , Jones L. 
Published: March 2020

20050 Background: The 88 kDa autocrine growth factor PC-Cell Derived Growth Factor (GP88) plays a critical role in breast tumorigenesis. GP88 expression was low in estrogen receptor positive cells, whereas in ER negative cells, it was constitutively overexpressed. Increased GP88 expression was associated with anti-estrogen therapy resistance in ER+ cells and Herceptin resistance in Her-2 overexpressing breast tumors. Antisense inhibition of GP88 expression in human breast adenocarcinoma lead to inhibition of tumor growth in vivo. Immunohistochemical studies have shown that GP88 was expressed in 80% of invasive ductal carcinomas in correlation with expression of poor prognosis markers whereas normal tissues and benign breast lesions were negative. Since GP88 is secreted by breast cancer cells, we examined whether GP88 was found in the circulation at an elevated level in the sera of breast cancer patients when compared to healthy individuals. Methods: A blood sampling study was conducted to determine the serum level of GP88 in healthy volunteers (HV) and breast cancer patients (BC pts). Ten ml of blood was drawn every three months to obtain serum. GP88 serum concentration was determined in triplicate by quantitative enzyme immunoassay using human GP88 as standard. 126 BC pts were accrued. . In addition, sera from 53 healthy volunteers were obtained to establish a GP88 baseline in HV. BC pts characteristics: race: Caucasian- 61, African American-60, Asian-5; median age: 52.5 (range 26–84), stage I-32, II-34, III-18, IV-42. Results: Circulating GP88 was measurable in the serum. Median level of GP88 was 32.8 ng/ml (range 15.3–42.8) in HV and 43.8 ng/ml (range 15.4–158.4) in BC pts, (p-value = 0.0007). Conclusions: GP88 is measurable in the sera of HV and BC pts. Comparison between the two groups indicates that GP88 level is significantly higher in the sera of BC pts. These studies are important since it identifies GP88 as a measurable biomarker that is also a therapeutic target of malignant transformation or malignant progression of breast carcinoma (BC). Future studies will examine the correlation of GP88 level with BC prognostic factors. Correlation between the serum level of GP88 and therapeutic response to systemic therapy in breast cancer patients will also be assessed. [Table: see text]
geNorm assessment of the efficacy of novel reference genes ALG9, TFCP2 and ZNF410 compared to GAPDH for real time normalization in GI carcinoids
Authors: Eick G. , Kidd M. , Mane S. , Nadler B. , Champaneria M. , Pfragner R. , Modlin I. M. 
Published: March 2020

20052 Background: Robust quantitation of potential clinical marker genes using quantitative real-time PCR (Q RT-PCR) is critically dependent on accurate normalization. Although GAPDH has historically been used for normalization, its expression has been shown to vary widely between different tissues and experimental conditions. Additionally, conventional normalization strategies based on a single housekeeping gene can lead to large normalization errors. The determination of a panel of genes that have robust expression in the experimental system being studied is therefore essential to ensure accurate normalization and interpretation of results. Methods: Based upon the availability of large-scale gene databases, we developed methodology to identify highly expressed genes (mean log-transformed expression levels: 4–8 in all samples) with low variability (S.D. < 0.22) in a Affymetrix U133A dataset of 36 gastrointestinal tumors and normal tissues. Eight novel candidate reference genes were identified and their expression levels established by Q RT-PCR in an independent set of GI tissue samples (n = 24). The geNorm tool was used to identify the most stably expressed set of genes amongst the 8 candidate genes. The expression levels of 3 potential GI tumor marker genes, namely the adhesin MAGE-D2, the metastasis-associated MTA1, and the mitosis regulator, NAP1L1, were normalized to GAPDH or geNorm and compared. Results: geNorm identified 3 genes, ALG9, TFCP2 and ZNF410, as the most robustly expressed control genes. GAPDH, in contrast, exhibited the highest variability and was considered the least stable gene of the nine evaluated. Two previously-identified target genes, MAGE-D2 and MTA1, were significantly elevated (p < 0.05) in malignant tumor samples (vs normal GI samples) when normalized by geNormATZ but not when normalized using GAPDH. NAP1L1 was only over-expressed in small intestinal carcinoids (normalized to geNormATZ). Expression levels of this gene were high in normal gastric mucosa. Conclusions: We provide a robust basis for the establishment of a reference gene set using GeneChip data and provide evidence for the clinical utility of identifying reference genes using the geNorm approach in GI neuroendocrine tumors. No significant financial relationships to disclose.
What is the role of QTc prolongation assessment in new drugs development phase I oncology trials?
Authors: Curigliano G. , Cipolla C. , Sessa C. , Noberasco C. , De Pas T. , Colombo N. , Spitaleri G. , Putzu C. , Milani A. , De Braud F. 
Published: March 2020

2006 Background: QT interval prolongation associated with “torsades de pointes” (TdP) has been a common cause of withdrawal from the market for several promising drugs. We determined the degree of QT prolongation in patients treated within a phase I study with a thioxanthone derivative known to have arrhythmogenic potential. Methods: Clinical data and serial ECGs from 31 patients with advanced tumors who received 86 courses of investigational drug were prospectively reviewed. The drug has been administered intravenously over 24 hours every 3 weeks. Patients have been on a 24 hour Holter monitor until 24 hours after infusion was complete. Three baseline ECGs were done and compared to those every 6 hours during therapy and once 6 hours after the infusion was complete. ECGs were read at a central lab according to a standard protocol. All QT measurements were then corrected for heart rate (QTc) using Bazett’s formula (QTc = QT interval divided by the square root of the R-R interval). Results: Overall,843 ECG tracings were obtained, all evaluable for analysis. No basal ECG showed significant abnormalities. Prolonged QT intervals developed in 2 patients without clinical symptoms (1 patient had intervals 500 milliseconds).In both cases it was associated with the maximum concentration of the drug. Compared with baseline, the QTc interval was prolonged by 30 to 60 milliseconds in 20% of total tracings, and by more than 60 milliseconds in 2% of ECGs. In patients receiving multiple courses, QTc intervals returned to pretreatment levels before the second course. Conclusions: The assessment of QTc prolongation was a major effort in this study but produced an accurate information about such event. In phase I study such an effort is justified when arrhythmogenicity is suspected. The timing of collection of ECGs should be guided by the available preclinical information about the pharmacokinetic profile of the drug. Nevertheless uncertainty remains regarding the specific relationship between the degree of QT prolongation and the risk of life-threatening arrhythmias. The decision to use the drug ultimately has to be based on an estimation of the perceived risk relative to expected benefits for patients. No significant financial relationships to disclose.
Survival in women with breast cancer having children with pediatric tumors—-A nationwide cancer register study
Authors: Olsson H. , Bladström A. 
Published: March 2020

20071 Background: Breast cancer is a heterogenous disease with a minoruty of cases having a hereditary background. Hereditary factors could affect disease prognosis and it has been postulated that breast cancer patients having p53 germline mutations have an especially bad prognosis. In the present study survival has been compared in women with breast cancer with and without having a child with a pediatric tumour. Methods: Using and linking the population based fertility registry and cancer registry the offspring of all women with breast cancer diagnosed between 1961–1999 was traced in Sweden. All women with breast cancer with a pregnancy were included. Further all childhood cancer cases < 21 years of age were identified among the offspring. Likewise using census data the women’s overall survival was followed. Results: Patients with breast cancer having a child with a childhood malignancy (n = 254) had a significant worse survival compared with other women with breast cancer (n = 74,781), RR = 1.25, 95% CI 1.02–1.55, p < 0.04, adjusted for age at diagnosis, year of diagnosis, parity and time since last pregnancy. Mothers with children with pediatric sarcomas or acute myeloid leukemias seemed to be strongest associated with a worse survival in the mother (RR = 1.38 and RR = 1.69). The worse survival of the mother was present regardless if the breast cancer diagnosis was before or after 50 years of age. Conclusion: The results suggest that genetic syndromes such as the Li-Fraumeni syndrome and possibly other childhood/adult cancer syndromes if present worsens the survival for breast cancer patients. This familial factor may be of importance for future treatment stratification. It is unknown whether other tumor diseases in the mother may be associated with a worse survival if their children have had cancer. No significant financial relationships to disclose.
Validation of clinical prognostic factors for survival in 1,043 Japanese patients with metastatic renal cell carcinoma (mRCC)
Authors: Tomita Y. , Naito S. , Ozono S. , Terachi T. , Naito S. 
Published: March 2020

20075 Background: Increasing number of patients and death of RCC has been reported in many countries including US and Japan. For choosing optimal treatment, determining risk of patients is essential. Although several prognostic factors of mRCC has been investigated, it was based on data of Caucasian. Since racial difference in biological character of cancer has been identified in several types of cancer, we validated the relationship between reported prognostic factors and survival in patients with mRCC in Japanese patients. Methods: We reviewed the medical records of 1,043 patients who had mRCC in the period to follow up, or at the first visit examination between 1988 and 2001 of 26 center in Japan. Results: Average survival of the 708 patients who died for renal cell carcinoma was 20.84 months. On the other hand, histological high grade (> WHO grade 2), metastasis at the first visit examination, short period from the first visit to recognition of metastasis (within 12 months), brain, bone or, liver metastasis, a number of metastatic sites (>1), high performance status (ECOG PS >0), high corrected serum calcium, low hemoglobin, and high CRP were prognostic factors for survival. However, LDH were not related. The median time to death with zero risk factor was 38.5 months, but those with >3 risk factors was 10 months. The patients with >4 risk factors was 4 months. Conclusions: Almost all prognostic factors reported in previous studies mainly based on Caucasian data had same impact on Japanese patients with mRCC. However, elevation of LDH did not have relationship to prognosis. Moreover, the time to death from diagnosis of mRCC in patients with poor prognostic factors was likely shorter compared with other studies. These results suggest Japanese with mRCC might have different character from those in US or European countries though further study in patient’s and tumor characteristics is warranted. No significant financial relationships to disclose.