Influence Cancer Development Research Articles

DETECTION OF EPSTEIN-BARR VIRUS GENOME IN ORAL CAVITY SQUAMOUS CELL CARCINOMA SAMPLES OF RUSSIAN PATIENTS

Oral cavity squamous cell carcinoma (OC-SCC) is the most common and aggressive malignancy of the oral cavity. Recent studies have revealed infections with human papilloma virus (HPV) as an additional risk factor for oral squamous cell carcinoma development, while distinguished role of Epstein-Barr virus (EBV) remains still uncertain. However, the evidence for association between virus infection and risk of oral squamous cell carcinoma is controversially and varies significantly by geographic regions and race. The aim of the present study was to elucidate the prevalence of HPV and EBV in OC-SCC samples of Russian patients from Moscow region. We investigated fresh-frozen tumor tissue fragments obtained from 11 patients with OC-SCC. DNA was extracted and the viral genome was examined by quantitative PCR assays with highrisk type-specific HPV and EBV specific markers followed by sequencing-based analysis. No HPV infection in analyzed OC-SCC samples was observed, while EBV was identified in 70.0% (7/10) of patients. Further based on Q-PCR amplification of the EBV targets including BamHI-W, EBNA1 and C-terminal fragment of LMP1 gene, EBV infection and measurement of virus load in the tumor samples was assessed. Sequencing LMP1-positive products revealed that the most samples (5/6) contained variants LMP1 with Cao deletion characterized by an increased transforming potential. These data suggest that prevalence of EBV infections is common and may influence cancer development, although detected LMP1 variants of EBV are not necessarily associated with the pathogenesis of OC-SCC. Further studies are necessary to determine the potential role of EBV and its possible importance as an infection factor in OC-SCC.

Ligand-activated progesterone receptor B activates transcription factor EB to promote autophagy in human breast cancer cells

Autophagy is an evolutionary conserved, self-eating process that targets cellular constituents for lysosomal degradation. Transcription factor EB (TFEB) is a master regulator of autophagy by inducing the expression of genes involved in autophagic and lysosomal degradation. In breast cancer, ligand-activated progesterone receptor has been reported to influence cancer development by manipulating the autophagy pathway. However, understanding of the mechanism that underlies this autophagic response remains limited. Herein, we report that prolonged treatment with progestin R5020 upregulates autophagy in MCF-7 human breast cancer cells via a novel interplay between progesterone receptor B (PRB) and TFEB. R5020 upregulates TFEB gene expression and protein levels in a PRB-dependent manner. Additionally, R5020 enhances the co-recruitment of PRB and TFEB to each other to facilitate TFEB nuclear localization. Once in the nucleus, TFEB induces the expression of autophagy and lysosomal genes to potentiate autophagy. Together, our findings highlight a novel functional connection between ligand-activated PRB and TFEB to modulate autophagy in MCF-7 breast cancer cells. As breast cancer development is controlled by autophagy, the progestin-PRB-TFEB transduction pathway warrants future attention as a potential therapeutic target in cancer therapy.

The μ-opioid receptor (MOR) promotes tumor initiation in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the most prevalent subtype of liver cancer. Anesthetic regimens possibly influence cancer development. Exploration of novel, effective targets for liver cancer is the current hotspot in cancer treatment. A previous study conducted by us has demonstrated that enhanced expression of the μ-opioid receptor (MOR) promotes cell proliferation, adhesion, migration, and tumorigenesis. The current study investigates whether MOR regulates self-renewal of hepatocellular carcinoma stem cells (HCSCs). We utilize cell function assays, siRNA, shRNA, flow cytometry sorting, and other molecular biology techniques for this purpose. The results indicate that MOR expression is positively related to hepatocarcinoma progression. Silencing MOR greatly reduce HCC-related tumorigenesis both in vitro and in vivo and significantly extend the survival of tumor-bearing mice. Moreover, MOR silencing will greatly reduce colony formation by HCC cells, indicating down-regulation of cancer initiation. In conclusion, these results establish that MOR can be a novel and reliable HCSC marker and a potential therapeutic target against HCC via MOR-NFAT signaling.

Integration of microbiology, molecular pathology, and epidemiology: a new paradigm to explore the pathogenesis of microbiome-driven neoplasms.

Molecular pathological epidemiology (MPE) is an integrative transdisciplinary field that addresses heterogeneous effects of exogenous and endogenous factors (collectively termed 'exposures'), including microorganisms, on disease occurrence and consequences, utilising molecular pathological signatures of the disease. In parallel with the paradigm of precision medicine, findings from MPE research can provide aetiological insights into tailored strategies of disease prevention and treatment. Due to the availability of molecular pathological tests on tumours, the MPE approach has been utilised predominantly in research on cancers including breast, lung, prostate, and colorectal carcinomas. Mounting evidence indicates that the microbiome (inclusive of viruses, bacteria, fungi, and parasites) plays an important role in a variety of human diseases including neoplasms. An alteration of the microbiome may be not only a cause of neoplasia but also an informative biomarker that indicates or mediates the association of an epidemiological exposure with health conditions and outcomes. To adequately educate and train investigators in this emerging area, we herein propose the integration of microbiology into the MPE model (termed 'microbiology-MPE'), which could improve our understanding of the complex interactions of environment, tumour cells, the immune system, and microbes in the tumour microenvironment during the carcinogenic process. Using this approach, we can examine how lifestyle factors, dietary patterns, medications, environmental exposures, and germline genetics influence cancer development and progression through impacting the microbial communities in the human body. Further integration of other disciplines (e.g. pharmacology, immunology, nutrition) into microbiology-MPE would expand this developing research frontier. With the advent of high-throughput next-generation sequencing technologies, researchers now have increasing access to large-scale metagenomics as well as other omics data (e.g. genomics, epigenomics, proteomics, and metabolomics) in population-based research. The integrative field of microbiology-MPE will open new opportunities for personalised medicine and public health. Copyright © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

PO-146 Knockout validation of antibodies to components of the NF-kB signalling pathway

IntroductionNuclear factor-κB (NF-κB) plays a pivotal role in inflammation and innate immunity and aberrant regulation of NF-κB can influence cancer development and progression (Ref 1). As a result, a great deal of research is now focused on targeting this signalling pathway therapeutically. In order to accurately and reproducibly examine the NF-kB signalling cascade there is a need for highly specific antibodies. We, at Abcam, have developed knockout (KO)-validated antibodies targeted to distinct elements of the NF-κB signalling pathway to maximise experimental accuracy and reproducibility.Material and methodsOur KO validation program uses the extensive library of KO cell lines available from Horizon Discovery. Target genes are mutated via CRISPR-Cas9 within a haploid cell line, resulting in a frameshift and complete loss of gene expression. Target specificity is then confirmed via western blot, flow cytometry or immunocytochemistry. These KO cell lines function as true negative controls that can be used at a large scale to confirm antibody specificity to target proteins.Results and discussionsWe present antibody validation data for seven targets in the NF-κB signalling pathway. Of the 30 Abcam products tested, we demonstrate that 22 antibodies are specific. The remaining 8 have since been removed from our catalogue due to specificity concerns. We have now validated over 1000 antibodies using this KO method, including many that are relevant to immunology and immuno-oncology research.ConclusionFollowing validation with KO cell lines as a negative controls, we can recommend 22 highly specific antibodies to proteins in the NF-κB signalling pathway. KO validation is one of many strategies that we use to ensure the specificity of our antibodies. By providing robust and specific antibodies which work the first time, we hope to help researchers to improve the experimental reproducibility and accuracy of their studies.Reference1. Bastian Hoesel and Johannes A Schmid; Mol Cancer. 2013 12: 86. doi:10.1186/1476-4598-12-86

Abstract 1264: Commensal microbiota is involved in anti-PD-1 mAb-mediated antitumor activity in a preclinical tumor model

Abstract Background: A number of studies have shown that the gut microbiome may influence anti-tumor immune responses via innate and adaptive immunity, and the recent work in mice has highlighted the key roles of the gut microbiota in mediating tumor responses to chemotherapeutic agents and immunotherapies targeting PD-L1, PD-1 or CTLA-4. Multiple studies have demonstrated that the gut microbiome can markedly influence the outcome of PD-1 blockade in mice and patients and several commensal have been proved to be associated with favorable clinical outcome. Considering complexity of intestinal micro-organisms, a larger panel or wider spectrum of microbiota species maybe involved in this process. Methods: Since compositional differences in the microbiome may influence cancer development and responses to therapies, we sought to determine whether intestinal bacterial composition and abundances of the tumor bearing mice were associated with a specific treatment outcome of immunotherapy. To test this, we analyzed and compared an enrichment of intestinal microbiota of CT26 syngeneic tumor bearing mice before and after a series of anti-PD-1 mAb treatment were given. Results The results demonstrating that some strains were robustly enriched and this enrichment was associated with tumor response to anti-PD 1 therapy, while as other strains were down regulated upon the checkpoint blockade. Conclusions: Upon further characterization and confirmation, these intriguing findings could lead to identification of one or more microbiota strains that could have biomarker potentials or therapeutic values for cancer immunotherapy. Citation Format: Fei Chen, Jian Ding, Xiangchao Gu, Juan Zhang, wenqing Yang, Qian Shi. Commensal microbiota is involved in anti-PD-1 mAb-mediated antitumor activity in a preclinical tumor model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1264.

Interplay between regulation by methylation and noncoding RNAs in cancers.

Cancer is one of the most important health problems today; therefore, many researchers are focusing on exploring the mechanisms underlying its development and treatment. The field of cancer epigenetics has flourished in recent decades, and studies have shown that different epigenetic events, such as DNA methylation, histone modification, and noncoding RNA regulation, work together to influence cancer development and progression. In this short review, we summarize the interactions between methylation and noncoding RNAs that affect cancer development.

Postmenopausal Women with Endometrial Cancer Have Greater Metabolic Dysfunction and Higher BMI than Women with Benign Endometrium

Obesity is an independent risk factor for endometrial adenocarcinoma (EC), yet it is unknown which attributes of obesity contribute to EC pathogenesis. We hypothesize that heterogeneity in metabolic health distinguishes which obese women are at risk for EC. We previously found that more premenopausal women with pre-malignant hyperplasia had type 2 diabetes and dyslipidemia than women with benign hyperplasia (BH), despite similar BMI. BH is the ideal control condition since presenting symptoms are similar, yet malignant transformation is negligible. Thus, we sought to compare metabolic abnormalities in postmenopausal women with EC and BH. We reviewed medical records of 284 women who received a biopsy/hysterectomy at our center over 3 years. We examined non-Hispanic white women age 50-65 years with BMI 25-50 kg/m2, to limit variability in factors known to affect metabolism. 122 women had histologically confirmed EC (n=74) or BH (n=48), and 60% had lab data. Variables were analyzed by chi-squared or Student’s T test. Women with EC were older (58 ± 0.5 vs. 53 ± 0.4 years, p<0.0001) and had higher BMI (37 ± 1 vs. 32 ± 1 kg/m2, p<0.0001) than women with BH. BMI distribution was markedly different between groups, with a higher frequency and greater severity of obesity among women with EC (p<0.0001). More women with EC than BH had type 2 diabetes (19% vs. 4%, p<0.05) and hypertension (54% vs. 25%, p<0.01). Women with EC had higher triglycerides (140 ± 12 vs. 110 ± 13 mg/dL, p<0.05) and lower HDL (55 ± 2 vs. 60 ± 4 mg/dL, p=NS). However, these differences can be explained by the skewed BMI distribution. LDL and total cholesterol as well as statin use (20% vs. 25%) were similar between groups (p=NS). The striking differences in BMI among postmenopausal women with EC and BH highlights the role of obesity in EC pathogenesis. Since BMI affects metabolism, a prospective study of BMI-matched women is necessary to determine whether metabolic dysregulation independently influences cancer development. Disclosure K.M. Cooke: None. E. Dun: None. A.K. Anam: None. C. Flannery: None.

MicroRNA Networks in Breast Cancer Cells.

A variety of molecular techniques can be used in order to unravel the molecular composition of cells. In particular, the microarray technology has been used to identify novel biomarkers that may be useful in the diagnosis, prognosis, or treatment of cancer. The microarray technology is ideal for biomarker discovery as it allows for the screening of a large number of molecules at once. In this review, we focus on microRNAs (miRNAs) which are key molecules in cells and regulate gene expression post-transcriptionally. miRNAs are small, single-stranded RNA molecules that bind to complementary mRNAs. Binding of miRNAs to mRNAs leads either to degradation, or translational inhibition of the target mRNA. Roughly one third of all the mRNAs are postulated to be regulated by miRNAs. miRNAs are known to be deregulated in different types of cancer, including breast cancer, and it has been demonstrated that deregulation of several miRNAs can be used as biological markers in cancer. miRNA expression can for example discriminate between normal, benign and malignant breast tissue, and between different breast cancer subtypes.In the post-genomic era, an important task of molecular biology is to understand gene regulation in the context of biological networks. Because miRNAs have such a pronounced role in cells, it is pivotal to understand the mechanisms that underlie their control, and to identify how miRNAs influence cancer development and progression.

Mutational and epigenetic signatures in cancer tissue linked to environmental exposures and lifestyle.

In this article, we describe how recent advances in the study of mutational and epigenetic signatures in tumours provide new opportunities to understand the role of the environment and lifestyle in cancer development. Cancer-related mutational events have been investigated for decades but only recently the wide availability of genomic sequences and epigenomic data from thousands of cancer genomes has made it possible to identify numerous distinct mutational and epigenetic signatures through the application of advanced mathematical models. Some of these signatures have been linked to endogenous factors such as defective DNA repair or the action of APOBEC cytidine deaminases and to exogenous factors such as tobacco smoke, ultraviolet light, aflatoxins, aristolochic acid and ionizing radiation. More recently, it has been shown that exposure to factors such as tobacco smoke may also leave marks in the DNA methylation profile of both normal and tumour tissue in target organs. The analysis of mutational and epigenetic signatures is a novel and useful tool to study cancer. Their application to experimental studies and to studies with detailed data on environmental exposures and lifestyle is likely to improve our understanding of how the environment and lifestyle influence cancer development and its evolution.

Overview Of Public Health Dietary Guidelines For Prevention Of Cancer

Diet is now recognized as a vital element in cancer prevention strategies, both for its potential to influence cancer development directly and for its role in avoiding unhealthy adiposity. Dietary guidelines for cancer prevention based on systematic review synthesizing human and laboratory research identify choices that merit the highest priority. This review summarizes current understanding of diet’s multiple intersections with the process of cancer development and discusses major current dietary guidelines for cancer prevention. Dietary guidelines for reducing cancer risk address steps to provide sufficient amounts of cancer-protective factors and to limit exposure to foods and drinks that may facilitate cancer development while emphasizing the importance of an overall plant-rich dietary pattern and healthy weight. Tables and figures provide background to help health professionals discuss diet’s role in cancer prevention and emphasize priorities based on guidelines from systematic reviews of research rather than responding to single studies or non–evidence-based sources. These guidelines reflect the multiple roles that diet can play in cancer prevention and are compatible with dietary guidelines aimed at promotion of overall health. This review contains 4 figures, 3 tables and 58 references Key words: alcohol, American Institute for Cancer Research, cancer prevention, diet, dietary guidelines, dietary pattern, nutrition, obesity, plant-based diet, plant-rich diet, public health, recommendations, red and processed meat, reducing cancer risk, weight

Mutant p53 shapes the enhancer landscape of cancer cells in response to chronic immune signaling

Inflammation influences cancer development, progression, and the efficacy of cancer treatments, yet the mechanisms by which immune signaling drives alterations in the cancer cell transcriptome remain unclear. Using ChIP-seq, RNA-seq, and GRO-seq, here we demonstrate a global overlap in the binding of tumor-promoting p53 mutants and the master proinflammatory regulator NFκB that drives alterations in enhancer and gene activation in response to chronic TNF-α signaling. We show that p53 mutants interact directly with NFκB and that both factors impact the other’s binding at diverse sets of active enhancers. In turn, the simultaneous and cooperative binding of these factors is required to regulate RNAPII recruitment, the synthesis of enhancer RNAs, and the activation of tumor-promoting genes. Collectively, these findings establish a mechanism by which chronic TNF-α signaling orchestrates a functional interplay between mutant p53 and NFκB that underlies altered patterns of cancer-promoting gene expression.

Abstract 1293: ABO blood type and cancer risk: preliminary findings from a phenome analysis

Abstract Introduction: ABO blood type has long been implicated in disease susceptibility, including cancer. However, evidence for associations with many malignancies is mixed. We applied a novel phenome approach to test to cancer codes from electronic medical records (EMR) in relation to ABO blood type in a large predominantly Caucasian study population. Approach: Among adults aged 18-100, cancer case and control status were assigned using 58 general neoplasm related phenome codes to de-identified EMR at the Vanderbilt University Medical Center. Blood type from serologic assays was ascertained from EMR-linked laboratory reports. Associations between blood type and cancer phenomes were quantified with Odds Ratios (OR) and corresponding 95% Confidence Intervals (CI) from logistic regression in models adjusted for sex and stratified by race/ethnicity. Only analyses with at least 100 cases per strata were conducted. Results: Among 221,015 Non-Hispanic Caucasians, 37,841 Blacks, 7,714 Hispanic Caucasians, and 3,616 Asian subjects with ABO blood type available in linked EMR, we evaluated 56, 37, 4, and 3 general cancer phenome codes, respectively. After employing Bonferroni corrections, ABO blood type was significantly associated with cancers of the pancreas, ovary, cervix, skin, and hematopoietic system. Caucasians with blood type O were less likely to have ovarian cancer (OR: 0.82, 95% CI 0.73-0.91) and pancreatic cancer (OR: 0.83, 95% CI: 0.74-0.92), and more likely to have squamous cell or other skin cancer (OR: 1.08, 95% CI: 1.04-1.13) and myeloid leukemia (OR: 1.15, 95% CI: 1.06-1.25) than those with other blood types (A, B, or AB). Hispanic Caucasians with blood type O were less likely to have cervical cancer (OR: 0.56, 95% CI: 0.38-0.82) than those with other blood types. No associations surpassed correction for multiple comparisons among Blacks or Asians. Conclusions: Our phenome approach confirmed known associations between blood type and risk of pancreatic and ovarian cancer, and adds to accumulating evidence supporting associations with skin cancer and leukemia. Our novel cervical cancer association among Hispanic Caucasians and other nominally significant findings, especially in understudied non-Caucasians, should be further evaluated in large and diverse populations. In addition, research to determine how ABO blood type may influence cancer development and progression, and if such associations can be exploited for risk prediction or cancer prevention is warranted. Citation Format: Alicia Beeghly-Fadiel, Ayush Giri, Lisa Bastarache, Jill Pulley, Jeremy Warner, Josh Denny. ABO blood type and cancer risk: preliminary findings from a phenome analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1293. doi:10.1158/1538-7445.AM2017-1293

Abstract 3706: Validation of human and mouse myeloid panels on the NanoString® nCounter® Platform

Abstract Introduction: The innate immune system profoundly influences cancer development as well as its response to various therapeutic approaches. Multiple myeloid cell lineages are central to the innate immune response during cancer and have been found to regulate diverse cellular processes encompassing immune suppression, angiogenesis, invasion, metastasis, and drug resistance. NanoString has collaborated with the laboratory of Prof. Lisa M. Coussens to develop research gene panels for quantitative and qualitative analysis of various myeloid subsets by evaluating relative enrichment of genes that mark recruitment and presence of select myeloid subtypes (e.g., granulocytes, dendritic cells, monocytes, macrophages, mast cells), their cell function, activation and effector status. Methods: Gene content for human and murine versions of the myeloid panels were developed from an in-house high content proteomic screen of macrophage activation (unpublished), genomic analysis of human tumor associated macrophages (unpublished), and data mining from publically available datasets featuring myeloid transcriptomes. In total, 692 genes were included in the human panel and 675 genes in the murine panel. The panels were optimized to work across a range of sample types including fresh frozen tissues, formalin-fixed paraffin-embedded (FFPE) samples, peripheral blood mononuclear cells and cell lysates. Results: To validate the panel content, we are profiling myeloid infiltrates in syngeneic murine models of mesothelioma and mammary tumorigenesis by flow cytometric analysis and the nCounter myeloid panels. Since the quality of the myeloid tumor response can skew toward adaptive immune activating (Th1) or suppressive (Th2) phenotypes, we will profile the same tumor samples with the NanoString PanCancer Immune Profiling panel to evaluate T cell activation status by gene expression and flow cytometry as well as global immune response. Finally, myeloid complexity and T cell activation status in both primary tumor and lung metastases will be evaluated by NanoString to assess regulation of adaptive immune responses by the innate compartment in distinct tumor microenvironments. Conclusions: Upon validation with these assays, the myeloid gene panels may provide insight into how the innate immune system regulates cancer development and response to therapeutic pressure. Improved characterization of the myeloid compartment may lead to greater insight into the fundamental biology of tumor-immune interactions, novel therapeutic strategies, and other applications. Citation Format: Sushil Kumar, Sarah Warren, Tiziana Cotechini, Terry Medler, Christina Bailey, Joseph M. Beechem, Joseph M. Beechem, Lisa M. Coussens. Validation of human and mouse myeloid panels on the NanoString® nCounter® Platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3706. doi:10.1158/1538-7445.AM2017-3706

The Human Microbiome and Cancer.

Recent scientific advances have significantly contributed to our understanding of the complex connection between the microbiome and cancer. Our bodies are continuously exposed to microbial cells, both resident and transient, as well as their byproducts, including toxic metabolites. Circulation of toxic metabolites may contribute to cancer onset or progression at locations distant from where a particular microbe resides. Moreover, microbes may migrate to other locations in the human body and become associated with tumor development. Several case-control metagenomics studies suggest that dysbiosis in the commensal microbiota is also associated with inflammatory disorders and various cancer types throughout the body. Although the microbiome influences carcinogenesis through mechanisms independent of inflammation and immune system, the most recognizable link is between the microbiome and cancer via the immune system, as the resident microbiota plays an essential role in activating, training, and modulating the host immune response. Immunologic dysregulation is likely to provide mechanistic explanations as to how our microbiome influences cancer development and cancer therapies. In this review, we discuss recent developments in understanding the human gut microbiome's relationship with cancer and the feasibility of developing novel cancer diagnostics based on microbiome profiles. Cancer Prev Res; 10(4); 226-34. ©2017 AACR.

Successful Chinese German Collaborations in Historical Dresden

Successful Chinese German Collaborations in Historical Dresden

Molecular Crosstalking among Noncoding RNAs: A New Network Layer of Genome Regulation in Cancer.

Over the past few years, noncoding RNAs (ncRNAs) have been extensively studied because of the significant biological roles that they play in regulation of cellular mechanisms. ncRNAs are associated to higher eukaryotes complexity; accordingly, their dysfunction results in pathological phenotypes, including cancer. To date, most research efforts have been mainly focused on how ncRNAs could modulate the expression of protein-coding genes in pathological phenotypes. However, recent evidence has shown the existence of an unexpected interplay among ncRNAs that strongly influences cancer development and progression. ncRNAs can interact with and regulate each other through various molecular mechanisms generating a complex network including different species of RNAs (e.g., mRNAs, miRNAs, lncRNAs, and circRNAs). Such a hidden network of RNA-RNA competitive interactions pervades and modulates the physiological functioning of canonical protein-coding pathways involved in proliferation, differentiation, and metastasis in cancer. Moreover, the pivotal role of ncRNAs as keystones of network structural integrity makes them very attractive and promising targets for innovative RNA-based therapeutics. In this review we will discuss: (1) the current knowledge on complex crosstalk among ncRNAs, with a special focus on cancer; and (2) the main issues and criticisms concerning ncRNAs targeting in therapeutics.

ASSOCIATION OF TESTOSTERONE WITH COLORECTAL CANCER (HT29), HUMAN GLIOBLASTOMA (A172) AND HUMAN EMBRYONIC KIDNEY (HEK293) CELLS PROLIFERATION.

Testosterone influences cancer development. This in vitro experiment was exerted to determine the association of testosterone with human colorectal cancer(HT29), glioblastoma (A172) and human embryonic kidney(HEK293) cells proliferation. HT-29, A172 and HEK293 cell lines were cultured in standard growth medium, then randomly divided into control group (not exposed to testosterone) and groups exposed to 1, 10, 100 and 1000 μg/mL of testosterone. Cell viability was quantified by MTT assay. Statistical analysis was performed using ANOVA. Viability of HEK293 cells significantly increased in groups exposed to 1 μg/mL and decreased in groups exposed to 100 and 1000 μg/mL of testosterone compared to control group (P<0.05, P<0.05 and P<0.001, respectively). Viability of HT29 cells significantly increased in groups exposed to 10 and 100 μg/mL of testosterone and significantly decreased when exposed to 1000 μg/mL of testosterone compared to control group (P<0.05, P<0.001 and P<0.001, respectively). Viability of A172 cells significantly decreased in groups exposed to 100 and 1000 μg/mL of testosterone compared to control group (P<0.001). In conclusion, different doses of testosterone have enhancing or suppressive effects on HEK293, HT29 and A172 cells proliferation; according to which, considering clinical use of testosterone therapy for cancer treatment is a highly controversial issue.

A glyco-competitive assay to demonstrate the stochasticity of fate decisions in Escherichia coli

Abstract Interaction bacteria-gut, via glycan associations, contribute to the selection of microbial communities along the gastrointestinal tract, influencing cancer development. The mechanism causing microbiome alterations is unknown, while this understanding would be pivotal to identify medical therapies. The molecular associations between Escherichia coli bacteria and glucose, both in solution and immobilized at the surface, were studied showing the dependence of E. coli glucose binding on the sugar form. Classical kinetic models were used to derive the reaction equilibrium and adsorption constants, 8 mM −1 and 1 (cell/mL) −1 and to explain the uptake. E. coli preferred the free glucose, whereas in a deprived environment, the anchored glucose became the major source of carbon for the bacteria. A stochastic algorithm disclosed that after initial transient, E. coli privileged the anchored glucose rather than the free sugar, independently on the concentration. The biochemical approach alone failed to describe the effective behavior of the cells and that several parameters can affect the behavior of the bacteria. From this result, more sophisticated models of the destruction of the gut barrier can be derived, such as the mechanism whereby E. coli can switch the immune system on and off to cause cancer and its metastasis.

Human Milk and Matched Serum Demonstrate Concentration of Select miRNAs.

Pregnancy-associated breast cancers (PABCs), especially those diagnosed after childbirth, are often aggressive, with a poor prognosis. Factors influencing PABC are largely unknown. Micro(mi)RNAs are present in many human body fluids and shown to influence cancer development and/or growth. In six nursing mothers, we determined if breast cancer-associated miRNAs were (1) detectable in human breast milk and (2) if detectable, their relative expression in milk fractions compared to matched serum. We evaluated by quantitative PCR the expression of 11 cancer-associated miRNAs (10a-5p, 16, 21, 100, 140, 145, 155, 181, 199, 205, 212) in breast milk cells, fat and supernatant (skim milk), and matched serum. miRNA expression was detectable in all samples. For 10/11 miRNAs, mean relative expression compared to control (ΔCt) values was lowest in milk cells, the exception being miR205. Relative concentration was highest in the skim fraction of milk for all miRNAs. Expression was higher in skim milk than matched serum for 7/11 miRNAs and in serum for 4/11 miRNAs. miR205 expression was higher in all milk fractions than in matched serum. In conclusion, the expression of breast cancer-associated miRNAs is detectable in human breast milk and serum samples. The concentration is highest in skim milk, but is also detectable in milk fat and milk cells.