bluuebunny/crossref_metadata_2025_split huggingface dataset:
miR-221 to modulate tumor growth in vivo and as a regulator of TGF.
Authors: Spahn Martin , Zoni Eugenio , Krebs Markus , Herreiner Philip , Kalogirou Charis , Chen Lanpeng , Snaar-Jagalska Ewa , Thalmann George N. , Kruithof-de Julio Marianna , Riedmiller Hubertus , Kneitz Burkhard 
Published: September 2018

e23008 Background: Despite the advances in cancer therapy, when Prostate Cancer (PCa) progress to castration resistant phase, patients develop incurable bone metastases. Understanding the processes that regulate homing and survival of metastatic cancer cells in the bone is crucial for the identification of new therapies. TGF-β signaling plays a major role in bone remodeling and according to the “vicious cycle hypothesis” is a master regulator of maintenance of prostate cancer cells in lytic bone lesions. microRNAs (miRs) are a class of small non-coding RNAs that regulates many biological process. miR-221 expression has been previously associated with prostate cancer progression. Here we studied the effect of miR-221 on TGF-β signaling and the impact of miR-221 on tumor growth in vivo Methods: miR-221 was overexpressed in PC3 and RWPE-1 cells and PMEPA was monitored by RT-qPCR and western blot. Expression of miR-221 in PC3 and RWPE-1 cells was assessed by RT-qPCR. Luciferase assay was used to investigate the interaction between miR-221 and PMEPA1. For zebrafish experiment, fluorescently labelled PC-3M-Pro4 cells overexpressing miR-221 were injected in the Duct of Cuvier (DC) of zebrafish embryos. Results: miR-221 overexpression resulted in decreased PMEPA1 mRNA and protein in PC-3 cells. Luciferase reporter assay indicated that miR-221 can directly interact with PMEPA1 3’ UTR. We show an enhancement of the proliferative effect of TGF-β in PC-3 cells, following miR-221 overexpression. In conclusion, we observed increased Smad2 activation upon TGF-β treatment in miR-221 overexpressing PC-3 cells. Inoculation of PC-3M-Pro4 cells overexpressing miR-221 in the DC of zebrafish embryos resulted reduced tumor burden compared to control. Finally, we observed inverse correlation between miR-221 and PMEPA1 expression in normal vs. tumor tissue collected from PCa patients. Conclusions: Our results indicate that miR-221 is a regulator of TGF-β signaling via modulation of PMEPA1 and miR-221 overexpression can reduce tumor growth in vivo.
Low T3 syndrome with hyperthyrotropinemia and early detection of liver metastasis.
Authors: Kaplieva Irina V. , Frantsiyants Elena Mikhaylovna , Trepitaki Lidia K. , Pogorelova Yulia A. , Bandovkina Valeria , Tkalya Lyudmila Dmitriyevna , Islamova Elena F. , Cherkes M. A. , Kit Oleg I. 
Published: September 2018

e23010 Background: Numerous clinical studies have shown an association between changing thyroid gland (TG) function and the development of tumors. The purpose of the study was to analyze the dynamics of thyroid hormones in the pituitary gland, TG and blood serum (BS) during liver metastasis in order to create thyroid profile of metastasis and to detect thyroid markers of metastasis in BS. Methods: The experiment included 44 white male rats weighing 180-250 g. Sarcoma 45 was transplanted intrasplenically. Levels of thyroid stimulating hormone (TSH) were studied in the pituitary gland, TG, BS; the amount of free (fT4) and total (T4) thyroxine and free (fT3) and total (T3) triiodothyronine in TG and BS were determined by radioimmunoassay (Immunotech, Czech Republic; Arian analyzer, Russia). Results: Exertion of the pituitary gland and overproduction of TSH were observed since the early days of the tumor development, later accompanied by TG hypofunction. Quantitative changes of thyroid hormones in organs did not coincide with their dynamics in BS. The first diagnostic signs of experimental liver metastasis, in the absence of formed metastases in the body, were hyperthyrotropinemia and a tendency to fT3 decrease in BS. Already formed liver metastases were characterized by a marked low T3 syndrome which transformed into a more severe lowT3/lowT4 syndrome during the secondary metastasis. Conclusions: The development of liver metastases is accompanied by the exertion and imbalance of the thyroid system function. Analysis of dynamics of the thyroid axis hormones in BS allows predicting the appearance of liver metastasis and identification of “the point of no return” in the development of the pathology that leads to secondary metastasis and irreversible progression of the disease.
A novel role for 17-Β estradiol and testosterone in osteosarcoma tumorigenesis and metastasis.
Authors: Lichterman Jake N. , Post Robert , Menken Luke , Saad Bishoy , Delgado Justin , Sall Lauren , Karr Matthew , Kozlowski PIotr , Degenhardt Kurt 
Published: September 2018

e23012 Background: Pulmonary metastases still remains the leading cause of mortality in osteosarcoma, with a 5-year survival rate of less than 30%. 17-β estradiol and testosterone are known to promote tumorigenesis and metastasis in other cancers, but its role in osteosarcoma tumorigenesis and metastasis is unclear. An understanding of the mechanism that these hormones have in osteosarcoma tumorigenesis and metastasis will lead to new therapeutic strategies using currently available targeted therapies used in breast and prostate cancer. Methods: We began by assessing the in vitro characteristics of two human osteosarcoma cell lines treated with 17-β estradiol or testosterone via proliferation and invasion assays. For the in vivo experiments, NCR nu/nu mice were injected with two luciferase-tagged human osteosarcoma cell lines and treated with 17-β estradiol and testosterone slow-release pellets. We first injected the mice subcutaneously and monitored tumor growth via caliper measurements. In the second experiment we injected the cell lines intratibially and monitored primary tumor growth in the bone and monitored metastasis formation in the lungs via IVIS bioluminescent imaging. Quantification of luciferase signal (photons/second) in the lungs was performed via the Xenogen Living Image software. H&E and immunohistochemistry for luciferase expression was performed on lung tissue to quantify tumor burden in the lungs. Statistical analysis was performed using Graphpad Prism. Results: Treatment with 17-β estradiol and testosterone increased cell proliferation and invasion in a dose-dependent manner. We observed increased tumor growth with both 17-β estradiol and testosterone in the subcutaneous model. In the intratibial experiment, testosterone increases primary tumor growth, but has no effect on metastasis. 17-β estradiol decreases primary tumor growth, but increases metastasis in a cell line independent manner. Conclusions: This study demonstrates a novel role for the puberty-related sex steroid hormones, 17-β estradiol and testosterone, in osteosarcoma tumor growth and metastasis. Uncovering the mechanism behind this phenomenon will uncover new therapeutic options for this devastating disease.
Oncogenic microRNAs to predict relapse in early breast cancer patients.
Authors: Sochor Marek , Basova Petra , Pesta Michal , Bartos Jiri , Stopka Tomas 
Published: September 2018

e23021 Background: Early breast cancer is a frequent female disease with different outcomes. New approaches are needed in order to improve its prognosis. MicroRNAs (miRs) are modulators of gene expression and act as oncogenes and function to inhibit tumor suppressors and to promote metastasizing. Monitoring of miRs could be of benefit to the prognosis of EBC patients. Methods: We prospectively collected sera from 133 EBC patients (follow-up 53,25 months) in 3 time points (before I and after surgery II, after therapy III). Patients were stratified into high and low-risk groups according to HR, HER2, Ki-67, grade, LN. For RNA isolation serum was used followed by RT-qPCR and was applied longitudinal multivariate data analysis. Aim of the project was to determine serum expression of miR-155, miR-19a, miR-181b, miR-24 in 3 time points, to find difference in expressions between high and low-risk groups, to find association between miRs and clinical/pathological risk factors and associations in miRs expression and prognosis of EBC. Results: EBC patients significantly over-express miRs in time point I. In time point II the levels of miR-155, miR-181b, miR-24 significantly decreased ( p< 0.05). miR-19a decreased in time point III ( p= 0.00869). Levels of miR-155, miR-19a, miR-181b, miR-24 are significantly more abundant in high-risk in comparison to low-risk patients ( p< 0.05) and decrease upon therapy. The multivariate GEE model revealed that miR-155, miR-24 were predictive of the relapse ( p= 0.025 and 0.041). miR-19a, miR-181b are insignificant with respect to the relapse ( p> 0.05). Triple-negativity, HER2+, grade III, LN+ have no effect on the probability of relapse ( p> 0.05) when miRs are simultaneously taken into an account. The only risk factor that makes the prediction of relapse more precise is Ki-67 > 20% ( p= 0.013 in case of miR-155 and p= 0.023 in case of miR-24). Conclusions: OncomiRs are significantly more abundant in EBC patients at diagnosis and decline after therapy. Differences in miR levels reflect EBC risk groups. The data shows that miR-155 and miR-24 enable monitoring of EBC and predict relapse independently of clinical/pathological risk factors. Combining the miR levels with Ki-67 expression further specifies the relapse probability.
Difference in appearance of KRAS mutated circulating tumor DNA in patients with metastatic colorectal cancer during treatments.
Authors: Takayama Yuji , Suzuki Koichi , Ichida Kosuke , Fukui Taro , Watanabe Fumiaki , Kakizawa Nao , Muto Yuta , Kikugawa Rina , Hasegawa Fumi , Tsujinaka Shingo , Noda Hiroshi , Miyakura Yasuyuki , Konishi Fumio , Rikiyama Toshiki 
Published: September 2018

e23025 Background: Emergence of KRAS mutation in blood is observed in colorectal cancer patients who undergo chemotherapy, but its clinical significance is not well known. In this study, we focused on the difference in appearance of KRAS mutated circulating tumor DNA (MctDNA) and elucidated its association with treatments. Methods: Four hundred and fifty-one plasma samples were collected prospectively from 85 patients (pts) who underwent chemotherapy due to metastatic colorectal cancer in 2014 - 2016. Seven types of KRAS mutation in MctDNA were detected by droplet digital PCR creating oil droplets. To exclude false positive detection, mutation was validated. MctDNA amplified in oil droplets was selectively sorted by On-chip sorting system and mutation was determined by Sanger sequencing. Results: KRAS assessment in tumor tissues showed 29 pts with KRAS mutation (MT), 56 pts without KRAS mutation (WT). Among 29 pts with MT, KRAS assessment in plasma displayed 23 pts with MctDNA and 6 pts without MctDNA. The type of mutation in MctDNA was consistent with that detected in tumor tissues, indicating mutual exclusivity in KRAS mutation was confirmed. In 56 pts with WT, 28 pts showed MctDNA during treatments. Difference in appearance of MctDNA was recognized in several treatments. Gradual increase in detection of MctDNA was observed with anti-EGFR antibody, resulted in treatment resistance. Transient spike elevation was frequently seen in TAS-102, which associated with drug response. No specific appearance was recognized during treatments with other drugs including anti-VEGF antibody. MctDNA in oil droplets were successfully sorted even if a few droplets were targeted, and mutation was confirmed. Conclusions: Difference in appearance of MctDNA may associate with treatment response in patients with metastatic colorectal cancer during treatments. [Table: see text]
Circulating microRNAs (miRNAs) as predictors of relapse in early breast cancer (BC).
Authors: Agelaki Sofia , Papadaki Chara , Stratigos Michalis , Spiliotaki Maria , Markakis Georgios , Mastrostamatis Georgios , Mavroudis Dimitrios 
Published: September 2018

e23030 Background: Disease relapse is usually a lethal development following treatment of early BC. Recurrence could arise from a state of tumour dormancy during which there is growth restriction of undetectable micrometastases. The expression of dormancy and metastasis related miRNAs was evaluated in the plasma of patients (pts) with early BC obtained before adjuvant therapy in order to discover novel biomarkers for the prediction of relapse. Methods: PlasmamiR-21, miR-23b, miR-190, miR-200b and miR-200c expression was assessed by qRT-PCR in 133 pts with early BC (non-relapsed, n = 84; relapsed, n = 49). Expression was classified as high or low according to the median values. Results: No correlation was observed between the expression of miRNAs and the clinicopathological characteristics of pts. After a median follow-up of 94 mo, median Disease Free Interval (DFI) was significantly lower in pts with high compared to low miR-21 [105.03 mo vs not reached (NR); p= 0.001], miR-200c (105.2 mo vs NR; p= 0.007), or both miR-21 and miR-200c expression (81.37 mo vs NR; p= 0.001). miR-21-high was also associated with decreased median Overall Survival (OS; p= 0.041). In multivariate analysis the number of infiltrated axillary lymph nodes (N3 vs N0-N2, HR: 2.86; p= 0.004) and miR-21 expression (high vs low, HR: 2.824; p= 0.001) were independent negative prognostic factors for DFI, whereas negative hormone receptor status (HR: 3.062; p= 0.024) and miR-21 high (HR: 3.545; p= 0.029) independently predicted for worse OS. Moreover, miR-21 expression was higher in pts with early relapse (defined as relapse at ≤ 3 yrs) compared to non-relapsed at 5 yrs ( p= 0.032). In addition, higher miR-21 ( p= 0.038), miR-23b ( p= 0.039), miR-200b ( p= 0.027) and miR-200c ( p< 0.001) levels were observed in pts with late relapse (at ≥ 5 yrs) compared to those without relapse. Conclusions: Differential expression levels of metastasis and/or dormancy related circulating miRNAs are detectable before adjuvant therapy in pts with early BC presenting subsequent relapse compared to non-relapsed pts. Circulating miRNAs can predict for early or late recurrence years before clinical detection of metastases. These results merit prospective validation in an independent patient cohort.
Performance of Biocept's sample collection for tumor cell analysis.
Authors: Singh Veena M. , Sales Edgar V. , Huynh Lan , Vibat Cecile Rose T. , Barón Anna E , Feser William , Malinowski Heather , Merrick Daniel , van Bokhoven Adrie , Malkoski Stephen , Miller York E , Wolf Holly J , Varella-Garcia Marileila 
Published: September 2018

e23036 Background: Liquid biopsy is a minimally invasive and cost effective way to assess cancer biomarkers without the risk of surgical biopsy complications. Circulating tumor cell (CTC) analysis from body fluids can provide critical information towards early detection, prognosis and treatment decisions. Accurate CTC evaluations require optimal cell preservation. Cell lysis, DNA degradation, or membrane alterations compromise CTC analyses and accurate diagnoses. This work compares Biocept’s proprietary CEE-Sure BCT and Saccomanno's Cytology Fixative largely used for sputum collection. Methods: One million BT474 (HER2 amplified) or H3112 (ALK re-arranged) cells were spiked into 500 µl medium; 500 µl of CEE-Sure or Saccomanno fixative was added. Tubes were stored at 4°C for 1 day, 1 week, or 1 month. Cells were centrifuged, resuspended, and counted (Celigo). Around 150 cells in 15 µl of RPMI medium were flowed into Biocept's microfludic system for cell capture; recovery (%) was calculated. Captured cells were subjected to fluorescent in situ hybridization (FISH) analyses for qualitative signal evaluation. Results: As similar results were observed for both cell lines and all time points, combined data will be shown. Median cell recovery after CEE-Sure incubation was 14.1% (range 1.7–44%, n = 12) vs 5.4% (range 0.07–26.9%, n = 12) in Saccomanno's fixative. Median cell capture of ~150 cells fed into Biocept’s microchannel was 96% (range 72-98%) for CEE-Sure vs 82% (range 21-96%) for Saccomanno. Paired t-tests showed significant differences for both recovery and capture. FISH signals from CEE-Sure samples were qualitatively rated Fair to Good, while Saccomanno samples had Poor to Fair, grainy, non-specific signals. Conclusions: This preliminary work shows consistently higher cell recovery, better cell membrane maintenance, and higher quality FISH signals for samples stored in Biocept's CEE-Sure vs Saccomano’s fixative. With liquid biopsy testing gaining rapid traction, maximal cell stability during the transport and storage are crucial. Additional fixative comparison is ongoing in various patient specimen types. These results support expansion of molecular analyses in sputum samples enriched for lung epithelial cells.
Clinical significance of monitoring KRAS in the blood of pancreatic cancer patients.
Authors: Watanabe Fumiaki , Suzuki Koichi , Ichida Kosuke , Takayama Yuji , Fukui Taro , Kakizawa Nao , Endo Yuhei , Kaneda Yuji , Noda Hiroshi , Rikiyama Toshiki 
Published: September 2018

e23042 Background: KRAS monitoring provides valuable information for early diagnosis and prediction of treatment outcome in colorectal cancer. KRAS mutation is observed in only half of colon cancer patients, whereas it is detected in 90% of pancreatic cancer patients. Therefore, investigating tumor DNA in blood by KRAS monitoring may be even more valuable in pancreatic cancer patients. In this study, we elucidated the clinical significance of KRAS monitoring in pancreatic cancer patients during treatment. Methods: KRAS in tumor tissues was analyzed for mutations by Scorpion ARMS or RASKET in 38 patients with pancreatic tumors. KRAS in blood was analyzed for mutations (G12D, G12V, G12C, G12A, G12S, G12R, G13D, Q61L, and Q61H) using droplet digital polymerase chain reaction in 23 patients who underwent chemotherapy and who had KRAS mutation in their tissues. Results: KRAS mutation in tumor tissues was detected in 27 of 38 patients (71.1%). These 27 patients showed significantly poorer prognosis (24.4 months median overall survival time [MST]) than the six patients without mutation (p = 0.04), who were all alive. Patients with mutation of 12D in tumor tissues showed better prognosis than patients with other types of mutation; thus, mutation of 12D may have a different biological effect to other mutations. Monitoring of KRAS in blood revealed KRAS mutation in 14 of 23 patients (60.9%). These 14 patients exhibited significantly poorer treatment outcomes than patients without mutation. The MST of patients with KRAS mutation in their blood was significantly shorter than that of patients without KRAS mutation (17.7 months vs. 44.3 months, p = 0.03). Conclusions: KRAS mutation in blood could be a valuable predictive and prognostic biomarker in pancreatic cancer patients who undergo chemotherapy. Additionally, assessment of KRAS in tumor tissues may provide information about individual tumor biology.
Circulating tumor cell (CTC) as a significant prognostic factor in resected primary lung cancer.
Authors: Yoneda Kazue , Hashimoto Masaki , Takuwa Teruhisa , Matsumoto Seiji , Okumura Yoshitomo , Hasegawa Seiki , Tanaka Fumihiro 
Published: September 2018

e23044 Background: Circulating tumor cell (CTC) is a potentially useful marker in early diagnosis and monitoring therapeutic effects for patients with malignant tumors, but clinical significance of CTC in primary lung cancer remains unclear. We previously showed that CTC was a useful surrogate marker of distant metastasis in primary lung cancer (Clin Cancer Res 2009). In this study, we evaluated the prognostic value among completely resected patients after long-term follow-up. Methods: A total of 94 patients (median age, 68 years; 30 females and 64 males) who underwent complete resection for primary lung cancer (4 with small cell and 90 with non-small cell) were prospectively evaluated. At the time of enrollment into the study, 7.5mL of peripheral blood was sampled from each patient, and an EpCAM-based detection system (CellSearch) was used for detection of CTC. CTC was detected in 16 patients (CTC-positive, 14.9%). Results: CTC-positivity was significantly associated with a poor recurrence-free survival (5-year recurrence-free survival rate, 40% versus 72%; p<0.01) (Table 1), which was confirmed by a multivariate analysis (hazard ratio, 2.57 [95% CI, 1.26-5.26]; P=0.010). CTC-positivity was also associated with a poor overall survival (5-year recurrence-free survival rate, 62% versus 84%; p<0.05) (Table 1), which was confirmed by a multivariate analysis (hazard ratio, 2.76 [95% CI, 1.14-6.71]; P=0.025). Conclusions: CTC-positivity was associated with poor recurrence-free survival and poor overall survival in resected lung cancer. [Table: see text]
cfDNA mutation frequency in early stage breast cancer.
Authors: Kota Karthik Joshua , Gyanchandani Rekha , Rosenzweig Margaret Quinn , Brufsky Adam , Huggins-Puhalla Shannon Leigh , Miller Lori , Oesterreich Steffi , Lee Adrian V. 
Published: September 2018

e23047 Background: Though 93% of patient present with early stage breast cancer (EBC), metastatic recurrence is expected in 7-13%. To prevent estrogen receptor (ER) positive disease recurrence, endocrine therapy (ET) is given after local disease. ET guidelines recently extended therapy —specifically tamoxifen—from 5 years to 10 based on ATLAS and aTTom. This pilot study used blood samples from women 6 months post-ET to determine cell free DNA (cfDNA) mutation prevalence in disease-free patients Methods: Patients with EBC and continued follow-up after ET were recruited from the Magee Women’s Breast Cancer Clinic. Inclusion criteria were EBC, ER+, and completion of ET > 6 months prior to visit; the exclusion criterion was active disease. Patients gave 2 blood samples, placed in Streck and EDTA tubes and processed at 2 laboratory sites for cfDNA. Blood from patients with metastatic breast cancer (MBC) served as controls for mutation detection. cfDNA was amplified for ESR1 and PIK3CA genes. Targeted amplifications underwent digital droplet PCR to identify mutations: 4 for ESR1 (D538G, Y537C/N/S), 2 for PIK3CA (E545K, H1047R) Results: Ten EBC patients > 6 months post-ET (post-EBC; 2/5/3 of stage I/II/III) and 10 MBC patients gave samples. cfDNA yield between plasma isolated from EDTA and Streck tubes (including mutation allele frequencies) was not significantly different (p > 0.05); cfDNA yield from patients with MBC was > 2X higher than post-EBC (p < 0.001). MBC cfDNA had 2 monoclonal and 1 polyclonal (2 different) mutations in ESR1, while post-EBC cfDNA had none Conclusions: This pilot study shows cfDNA can be consistently isolated from Streck or EDTA-processed blood from patients with MBC and post-EBC; however, cfDNA levels are significantly higher in MBC. There were no mutations (ESR1 or PIK3CA) in post-EBC samples, though ESR1 mutations were found in MBC. Further studies are needed to determine if mutations in cfDNA can be found in patients without evidence of disease