bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tenascin C to regulate the function of sHER3, an endogenous inhibitor of neuregulin signaling.
Authors: Cai Chunlin , Zhu Mojun , Maihle Nita J. 
Published: September 2018

e23002 Background: HER3 expression is associated with poor prognosis in melanoma patients but the mechanistic basis has not been determined. An endogenous secreted isoform of HER3, p85-sHER3, has been shown by us to inhibit neuregulin-1 mediated, Akt-dependent breast and ovarian cancer cell growth in vitro. Here we demonstrate that p85-sHER3 also regulates neuregulin-dependent melanoma cell migration via an Akt- independent, tenascin C-dependent mechanism. Methods: Immunoblot analysis and RT-PCR were used to study HER3 and p85-sHER expression. Cell proliferation and chamber assays were conducted to determine the effect of p85-sHER3 on melanoma cell growth and migration. Mass spectrometry (MS) was performed to identify sHER3-interacting proteins. CRSPR in vitro editing methods were used to knock out tenascin C (TNC) expression. Results: HER3 and p85-sHER3 were expressed in 5 of 6 human melanoma-derived cell lines tested, with exception of SK-MEL-103 which did not express either gene product. Exogenous addition of 10 nM p85-sHER3 inhibited growth in 3 cells lines, although Akt phosphorylation was not reduced. Protein extracts from M14-MEL cell conditioned media were characterized by MS, and identified p85-sHER3 and TNC. Their association was confirmed by co-immunoprecipitation. sHER3 was further shown to inhibit neuregulin stimulated cell migration in 2 cell lines which exhibited high levels of TNC. CRSPR knockout of TNC in these cell lines (but not in control lines) eliminated sHER3 inhibition of neuregulin stimulated melanoma cell migration. Conclusions: Our results suggest that p85-sHER3, predominantly secreted by HER3-expressing melanoma cells, inhibits melanoma cell proliferation and migration via an Akt-independent mechanism, which is likely achieved through interaction(s) with TNC. TNC promotes an intermediate adhesive state favoring cell migration and this state appears to be mediated by Rho-associated kinase signaling. The Rho inhibitor, CCG-203971 has been shown to inhibit melanoma cell migration. Together, these results suggest the existence of a TNC and HER3-dependent signaling pathway in melanoma that regulates cell migration, and therefore may be correlated with poor patient survival.
Knockdown of TPPP3 to inhibit cell proliferation and invasion in human colorectal cancer.
Authors: Li Yintao , Yu Jinming 
Published: September 2018

e23006 Background: Tubulin Polymerization Promoting Protein Family Member 3, TPPP3, a member of the TPPP protein family, has been reported to play important roles in initiation and progression of human cancers, such as lung cancer. However, the expression and underlying function of TPPP3 in colorectal cancer (CRC) have not yet been fully clarified. Methods: In this study, the mRNA and protein levels of TPPP3 in 96 clinical CRC specimens were determined by RT-PCR and immunohistochemistry. The relation between TPPP3 expression and clinicopathologic characteristics and overall survival (OS) were evaluated. TPPP3 was stably knockdowned by shRNA. In addition, CCK-8、Colony formation、Flow cytometric、Transwell and Angiogenesis assay were to examine the biological function of TPPP3 in CRC cells in vitro. Results: We show that TPPP3 was significantly increased in CRC tissues and associated with aggressive factors and poor patient survival. Further experiments showed that knockdown of TPPP3 inhibited cell proliferation, migration and invasion and induced cell apoptosis in vitro. In addition, TPPP3 silencing resulted in a decrease of angiogenesis and S phase fraction. And TPPP3 significantly affected the invasion and migration of CRC cells via the expression of MMP-9, Rac-1 and E-cadherin. Conclusions: Our results suggested that TPPP3 played an important role in CRC progress and might serve as novel therapeutic targets for CRC treatment.
Characteristics of proteomic patterns in patients with metastatic and non-metastatic colorectal cancer.
Authors: Vodolazhsky Dmitriy I. , Kutilin Denis S. , Kharin Leonid , Kolesnikov Evgeniy N. , Bogdanova Olesia A. , Sustretov Vyacheslav A. , Trifanov Vladimir , Kit Oleg I. 
Published: September 2018

e23007 Background: Over the past 20 years, the number of deaths from colorectal cancer (CRC) increased by 46% worldwide: every year more than 1 million people are diagnosed with this pathology. Proteomic studies using protein microarrays are a very attractive strategy for the screening of new highly specific biomarkers which can improve prognosis prediction of CRC. We performed a screening proteomic study to find new informative biomarkers for metastatic and non-metastatic colorectal cancer which can be further used in routine histochemical studies. Methods: The samples of tumor and non-tumor tissues of 20 patients with histologically confirmed adenocarcinoma of the colon were studied. Extraction, labeling and hybridization of proteins on the slide containing 224 antibodies for key cellular proteins were performed using the Panorama Antibody Microarray - Cell Signaling Kit, and the GenePix 4100A scanner was used for scanning. The data were analyzed using GenePixTMPro 6.0. Results: We revealed a significant (p < 0.05) increase in expression of such proteins as C-myc, SMAD4, PCAF, Adaptin (b1 + b2), FAK Phospho (pY577), PKC g and Phospho-Pyk2 (pY579/580) by 2.1; 1.5; 1.5; 1.5; 2.7 and 1.6 times, respectively, in tumor tissue of patients with metastatic colorectal cancer compared to non-tumor colon tissues. A significant increase (p < 0.05) in expression of proteins NGFR p75 (Nerve Growth Factor Receptor) by 3.8 times and Phospho-Ta (pS199 / 202) by 1.9 times was found in tumor tissue of patients with non-metastatic CRC in comparison with non-tumor colon tissue. Conclusions: The study showed changes in proteomic profile of malignant colon tissue, and proteomic profile differed significantly in tumor tissues of patients with metastatic and non-metastatic cancer. We found changes in expression of 9 proteins, differential in each group of patients (with metastatic and non-metastatic CRC). These proteins have a high potential as biomarkers for CRC prognosis prediction.
The effects of very low doses of chemotherapeutic drugs on endothelial cells.
Authors: Sonugur Fatma Gizem , Ocal Muge , Babahan Cansu , Abdi Abgarmi Samira , Icli Fikri , Akbulut Hakan 
Published: September 2018

e23014 Background: Angiogenesis is one of the key processes leading tumor progression and metastasis. Conventional chemotherapeutic drugs are usually cytotoxic to tumor cells at the therapeutic doses. The lowered doses of those drugs have been shown to induce apoptosis in endothelial cells and implemented as metronomic schedule in the clinic. We aimed to study the effects of very low doses of chemotherapeutic drugs on endothelial cell functions and gene and protein expression of pro-and anti-angiogenic factors. Methods: We first determined the IC50 doses of 5-fluorouracil, irinotecan, and oxaliplatin in a human umbilical endothelial cell line (HUVEC). Then we’ve tested the effects of the IC50, -1log, -2log and -3log of IC50 doses on endothelial cell migration and tube formation. We’ve also studied the effects of the assigned doses on expression levels of the genes VEGF-A, VEGFR2, PDGF-B, angiopoietin-2 (Ang-2), endothelin-1 (End-1), thrombospondin-1 (Tbs-1), iNOS, eNOS by real-time PCR, protein secretion levels of VEGF-A, PDGF-BB, Ang-2, End-1 and Tbs-1 by ELISA and nitrate/nitrite production by a colorimetric assay. Results: The IC50 dose levels of 5-FU, irinotecan, and oxaliplatin were 9,2 μM, 9,8 μM and 11,8 μM, respectively. We found that all the -1log, -2log and -3log of IC50 doses of the drugs tested in the study significantly decreased the migration capacity of the HUVEC cells. However, no significant changes were observed in tube formation of endothelial cells. Likewise, the expression levels of pro-angiogenic genes including Ang-2, iNOS, eNOS, PDGF-B and VEGF-A were significantly decreased at the same doses of the drugs. Accordingly, the levels of Tbs-1 were significantly increased at all 3 dose levels of the drugs. The most efficacious doses of the drugs to decrease NO levels in HUVEC cells were -1logIC50 for 5-FU, -2logIC50 for irinotecan and -3logIC50 for oxaliplatin. Conclusions: In conclusion, our results suggest that 5-FU, irinotecan, and oxaliplatin could decrease the pro-angiogenic and increase anti-angiogenic factor production in endothelial cells at the doses much lower than the usual metronomic schedules and warrant further research in animal models and clinical trials as an anti-angiogenic treatment modality.
The expression of pluripotency genes regulates properties of cancer stem cells in MCF-7 breast cancer model.
Authors: Miftakhova Regina R. , Rakhmatullina Aigul R , Mingaleeva Rimma N. , Garanina Ekaterina E , Khaiboullina Svetlana F , Rizvanov Albert A 
Published: September 2018

e23018 Background: More than 1.6 million cases of breast cancer (BC) are diagnosed annually worldwide. Despite advances in diagnoses and treatment, BC remains the third leading cause of death in women. Metastases and chemotherapy resistance are the main factors contributing to BC mortality. The ability of tumor cells to overcome the drug-induced growth inhibition is now linked to a unique population of cancer initiating tumor cells, often referred as cancer stem cells (CSC). CSC represent a small fraction of tumor cells, therefore, currently used isolation protocols have a low yield and are poorly reproducible, hampering research on the role of these cells in cancer chemoresistance. We propose a novel approach to generate large quantities of CSC-like cells by genetic reprogramming of non-stem cancer cells. Methods: We postulate that CSC-enriched cell line can be developed in vitro by upregulating of proteins controlling cancer cell pluripotency including Oct4, Sox2, NANOG, KLF4 and c-Myc. Increased transcriptional and translational activity of these genes in MCF-7 cells was demonstrated by real time-PCR and Western blot. Proliferation and migration of cells overexpressing Oct4, Sox2, NANOG, KLF4 and c-Myc were analyzed as well. Also, changes in CSC population counts and their sensitivity to chemotherapy were investigated using sphere formation assay. Results: We found that cell proliferation rate was correlated with the expression level of c-Myc and Oct4. Increased CSC counts were found in cells with overexpressed Oct4 (62.7%), KLF4 (97%) and NANOG (121.3%) proteins as compared to the parental cells. Overexpression of SOX2, NANOG and KLF4 significantly increased CSC resistance to docetaxel (83,3±6,8; 93,3±4,5 and 80,3±5,0 spheres respectively) when compared to the original cells (48,3±3,0 spheres). Conclusions: We conclude that overexpression of pluripotency proteins Oct4, Sox2, NANOG, KLF4 and c-Myc changes the CSC counts and proliferation capacity of BC cells. Acknowledgements: The study was funded by RFBR, according to the research project No. 16-34-60210 mol_а_dk, and by Russian Government Program of Competitive Growth of Kazan Federal University.
Aspirin to inhibit the formation and EMT of circulating tumor cells in patients with metastatic colorectal cancer but not breast cancer.
Authors: Ni Chao , Yang Liu 
Published: September 2018

e23024 Background: High circulating tumor cells (CTCS) have been acknowledged as a poor indication in malignant disease. Besides, platelets are found play crucial role in tumor cells’ epithelial – mesenchymal transition and metastasis, so here we test if disruption of platelet function with aspirin, could decrease the number and inhibit the EMT of CTCs in metastatic colorectal (MCC) or breast cancer (MBC). Methods: Patients with MCC or MBC who were not receiving cytotoxic chemotherapy currently (except for capecitabine maintenance), while endocrine therapy, bisphosphonate or molecular targeted therapy were accepted. Base line of Platelet aggregation rate (PAR), CTCs’ number and molecular phenotype were recorded. Patients whose CTCs ≥ 5 cells/7.5ml peripheral blood were included and receive aspirin 100mg/day for the following 8 weeks. Then the phlebotomy was performed at 4, 8 weeks thereafter to obtain specimen and assess PAR, CTCs’ number and phenotype. The phenotypes of CTCs were classified into epithelial (E+), mesenchymal(M+) and middle type(express both E+ and M+ markers) with fluorescence in situ hybridization assay. Results: Forty patients (19 MBC and 21 MCC patients ) were enrolled. Base line CTC numbers were 8.7± 3.4 in MBC patients, the ratio of E+ and M+ type were 48.2 ± 22.4% and 25.1 ± 14.9% respectively; the base line CTC numbers were 10.7 ± 4.8 in MCC patients, and the ratio of E+ and M+ type were 45.3 ± 27.3% and 22.0 ± 18.5% respectively. Despite adequate platelets inhibition in both groups, CTC numbers were similar in MBC patients in the following 8 weeks ( p= 0.0532 ), while the fraction of E+ or M+ CTCs were also unchanged (E+ CTCs p= 0.305; M+ CTCs p= 0.09); however, both CTC numbers and fraction of M+CTCs were markedly decreased in MCC patients (total numbers: p< 0.01; E+ CTCs p= 0.031; M+ CTCs p= 0.013) . Conclusions: Aspirin could decreased the number of CTCs and block EMT transition in MCC patients, and our results provide potential explanation of how aspirin impede the metastasis of colorectal cancer. But this effect could not be observed in MBC patients. Future studies evaluating the underlying mechanism of this differences remain of interest, and they may be informed by our results. Clinical trial information: NCT02602938.
Enumeration of circulating tumor cells and assessment of immunotherapy biomarker PD-L1 in patients with colorectal cancer.
Authors: Tseng Ju-Yu , Chen Chwen Cheng , Lin Chun-CHI , Lin Hung-Hsin , Chen Yen-Ru , Lee Chia-Ying , Wu Li-Fan , Wang Shin-Hang , Yu Hui-Min , Fang Wei-Feng , Jiang Jeng-Kai 
Published: September 2018

e23029 Background: Circulating tumor cells (CTC) play a prognostic role in patients with metastatic colorectal cancer (mCRC). Studies investigating detection of CTC in peripheral blood (PB) showed limited clinical sensitivity. This study compares clinical sensitivity of CTC analysis in mesenteric venous blood (MVB) vs PB. PD-L1 expression on isolated CTCs was also characterized in various stages of CRC patients. Methods: 8mL PB and MVB samples were collected into K2EDTA-containing vacationer tubes and subsequently performed CTC enumeration and PD-L1 analysis by MiSelect R system. CTC is defined as EpCAM+, cytokeratin+ and CD45- with intact nuclei. CTC and PD-L1+ CTC number were counted and correlated to the patients’ clinical manifestations. Results: Blood samples were collected from 75 CRC patients with various stages. In MVB, CTC detection rate was 20%, 33%, 44% and 58% for stage I, II, III, and IV respectively, and the overall detection rate is 41%. In PB, CTC only detected in sporadic cases (7%). Also, the amount of CTC detected in MVB was more abundant than that in PB ( P < 0.001) In addition, preoperative serum CEA levels were significantly associated with presence of CTC in MVB ( P= 0.011). PD-L1 expression was detected in 32% of the isolated CTCs, and was more abundant in those from late stage patients. The expression level of PD-L1 showed heterogeneity among CTCs. Conclusions: CTC enumeration in MVB could potentially increase the clinical sensitivity for CTC analysis in CRC. The CTC level in MVB significantly correlated with CEA levels which could serve as an important biomarker information for post-operative management. In additions, using MiSelect R System to determine PD-L1 expression on CTC could provide a promising approach for both patient selection or therapeutic monitoring of immunotherapies for CRC.
Landscape of somatic mutations in different subtypes: Advanced breast cancer with circulating tumour DNA analysis.
Authors: Yi Zongbi , Ma Fei , Guan Yanfang , Yuan Fang Li , Sun Xiaoying , Guan Xiuwen , Li Chunxiao , Yi Xin , Qian Haili , Xu Binghe 
Published: September 2018

e23039 Background: It is particularly important to provide precise therapies and understand tumor heterogeneity based on the molecular typing of gene expression. Yet the landscape of somatic mutations in different subtypes of advanced breast cancer (ABC) is largely undetermined. Methods: Overall, 100 blood samples were obtained from 100 advanced female breast cancer patients who underwent therapy at Cancer Hospital, Chinese Academy of Medical Sciences from March 2015 to September 2016. Mutations in 1021 tumor-related genes in ctDNA was assayed by gene-panel target-capture next-generation sequencing. Results: Somatic genomic alterations in ctDNA including copy number variants and point mutations were identified in 96 of 100 patients (96.0%). The number of somatic mutations varied markedly between individual patients (mean 2.9, range1-31). No difference was found between four subtypes for the number of somatic mutations. However, the mean number of somatic mutations was higher in age at 40-50 year than patients over age 60 ( p= 8.46 vs 3.88; p= 0.01). Results from multivariate analyses showed that the number of somatic mutations was increased with the number of endocrine therapy line ( p= 0.007). TP53 and PIK3CA were two most frequently mutated genes detected in ctDNA of 100 patients which were recurrently detected in 43 (43.0%) and 32 (32.0%) patients, respectively. ESR1/PIK3CA were more prevalent in HR+ cancers ( p= 0.007, 0.025 respectively). NOTCH1 are more frequently detected in HER2- group than in HER2+ patients (15.63% vs 2.94%; p= 0.033). In multiple logistic regression analysis indicated that pathological grade, tumour size at diagnosis and PR statue were positive associated with PIK3CA mutations. Multiple regression analysis also revealed that ki-67, metastatic at diagnosis, number of metastatic sites, number of endocrine line were associated with ESR1 mutations. Conclusions: The results revealed that different subtypes ABC have their own genetic alterations features. Certain gene mutations may be related to clinical treatment especially endocrine therapy.
Validation of the denaturing capillary electrophoresis (DCE) assay for non-invasive liquid biopsy in lung carcinoma: A study of concordance with the cobas EGFR mutation test v2.
Authors: Minarik Marek , Belsanova Barbora , Fiala Ondrej , Pesek Milos , Benesova Lucie 
Published: September 2018

e23040 Background: The detection of circulating tumor DNA (ctDNA), referred to as “liquid biopsy” is becoming a preferred option for some patients with lung adenocarcinomas. The key for a suitable technology is high sensitivity enabling to detect small fractions of mutated alleles, the method has to be solid against false-positives. The aim of this work was to compare our method based on capillary electrophoresis under denaturing conditions (DCE) to FDA and CE IVD approved commercial kit - cobas EGFR Mutation Test v2. Methods: A total of 44 plasma samples were acquired from patients with clinicaly confirmed stage IV lung adenocarcinoma. There were samples from patients prior to receiving therapy (with a known tissue EGFR genotype) as well as several ones showing progression on a previously administered antiEGFR-therapy (with no tissue info available at the time of plasma sampling). DNA was isolated from multiple 2 mL aliquots of each plasma. The first aliquot was run on cobas z480 real-time instrument using unmodified manufacturer's protocol. Another 2ml aliquot was run by DCE assay (accredited method under ISO 15189:2012), which is based on a PCR followed by a fragment analysis on an ABI capillary analyzer (Sanger sequencer). Results: The mutual concordance was 84.1% (37/44). cobas assay identified 11 EGFR-positive plasma samples (6 x Ex19del, 3 x L858R, 1x T790M combined with Ex19del and 1x T790M combined with L858R). DCE identified 8 EGFR-positive samples (6 x Ex19del, 1 x L858R, 1xT790M combined with L858R). DCE missed 5 of cobas-positive samples (2x Ex19del, 2x L858R and 1x T790M) while in turn identifying 2x Ex19del plasma samples that were concordant with tissue, but missed by cobas. Overall concordance with tissue was 73.3% (22/30) for cobas, 70.0% (21/30) for DCE assay and 80.0% (24/30) for both combined. Conclusions: DCE approach is reliable for low levels of mutations in plasma of lung cancer patients and comparable (and complementary) to cobas. Its advantage is simplicity, low cost and a universal, straighforward adaptability to virtually any mutation at any site allowing to detect oncogenic as well as tumor supressor mutations in ctDNA. Supported by grant 17-30748A.
Circulating LncRNAs as predictive biomarkers by molecular subtypes in advanced breast tumors.
Authors: Salvador Javier , Pecero Maria Luisa , Gil Ana , Rodriguez de la Borbolla Maria , Ruiz Manuel , Montano Alvaro , Gonzalez Rosario , Benavent Viñuales Marta , Molina Sonia 
Published: September 2018

e23041 Background: Breast cancer is the most common malignant tumor in women worldwide. The leading breast cancer deaths are due to advanced disease stage. For this reason, identification of novel breast cancer progression-associated biomarkers would enhance the clinical management of each patient. Long non-coding RNAs (LncRNAs) are defined as noncoding RNAs longer than 200 nucleotides and are involved in several regulatory processes, such as regulation of gene expression. Moreover, it has been described that LncRNA expression is deregulated in different human diseases such as breast cancer. However, it is unclear if circulating LncRNAs could be used as progression biomarkers of breast cancers. The purpose of this study was to evaluate the role of circulating LncRNAs in patients with advanced breast cancer. Methods: Expression of 84 LncRNAs was quantified by “RT2 IncRNA PCR Array Human Cancer PathwayFinder” (Qiagen) in plasma from 24 subjects (12 patients with advanced breast cancer and 12 healthy subjects). cDNAs were pooled into four groups, two groups containing a mixture of six patients with advanced breast cancer and another two groups containing a mixture of six cDNAs from healthy subjects. Three differentially expressed LncRNAs were selected for validation by specific real-time PCR assay. Results: Three deregulated LncRNAs were identified and validated in patients with advanced breast cancer versus healthy subjects. We found that LncRNAs GAS5 and ZFAS1 expression was decreased in the Luminal, Her2-positive and Triple Negative advance cancer groups compared to healthy subjects. In contrast, LncRNA RMRP expression was increased in Luminal and Her2-positive advance cancer groups. To our knowledge, there are no publications relating the LncRNA RMRP expression with advance breast cancer research. Conclusions: These results contribute to unreveal the role of circulating LncRNAs as biomarkers in the advanced breast cancer and provide novel non-invasive tools in this disease.