Differential effects of hypoxia on etoposide-induced apoptosis according to the cancer cell lines

Background: It is more and more recognized that hypoxia plays a role in the resistance of cancer cells to chemotherapy. However, the mechanisms underlying this resistance still need deeper understanding. The aim of this study was to investigate the effect of hypoxia on this process since hypoxia is one of the hallmarks of tumor environment. Results: The effect of hypoxia on the apoptosis induced by etoposide, one drug commonly used in chemotherapy, was investigated using three different cancer cell lines. Gene expression changes were also studied in order to delineate the mechanisms responsible for the hypoxia-induced chemoresistance. We observed that hypoxia differentially influenced etoposide-induced cell death according to the cancer cell type. While hypoxia inhibited apoptosis in hepatoma HepG2 cells, it had no influence in lung carcinoma A549 cells and further enhanced it in breast cancer MCF-7 cells. Etoposide increased p53 activity in all cell lines while hypoxia alone decreased it only in HepG2 cells. Hypoxia had no influence on the etoposide-induced p53 activity in A549, increased p53 abundance in MCF-7 cells but markedly decreased p53 activity in HepG2 cells. Using low density DNA arrays to detect the expression of genes involved in the regulation of apoptosis, etoposide and hypoxia were shown to each influence the expression of numerous genes, many of the ones influenced by etoposide being p53 target genes. Again, the influence of hypoxia on the etoposideinduced changes was different according to the cell type. Conclusion: These results evidenced that there was a striking parallelism between the effect of hypoxia on the etoposide-induced p53 stabilization as well as p53 target gene expression and its effect on the etoposide-induced apoptosis according to the cell type. They are very interesting not only because they provide one possible mechanism for the induction of chemoresistance under hypoxic conditions in cells like HepG2 but also because they indicate that not all cell types respond the same way. This knowledge is of importance in designing adequate treatment according to the type of tumors

Hypoxia induces protection against etoposide-induced apoptosis: molecular profiling of changes in gene expression and transcription factor activity

Background: it is now well established that hypoxia renders tumor cells resistant to radio- but also chemotherapy. However, few elements are currently available as for the mechanisms underlying this protection. Results: in this study, physiological hypoxia was shown to inhibit apoptosis induced in HepG2 cells by etoposide. Indeed, hypoxia reduced DNA fragmentation, caspase activation and PARP cleavage. The DNA binding activity of 10 transcription factors was followed while the actual transcriptional activity was measured using specific reporter plasmids. Of note is the inhibition of the etoposideinduced activation of p53 under hypoxia. In parallel, data from low density DNA microarrays indicate that the expression of several pro- and anti-apoptotic genes was modified, among which are Bax and Bak whose expression profile paralleled p53 activity. Cluster analysis of data unravels several possible pathways involved in the hypoxia-induced protection against etoposide-induced apoptosis: one of them could be the inhibition of p53 activity under hypoxia since caspase 3 activity parallels Bax and Bak expression profile. Moreover, specific downregulation of HIF-1α by RNA interference significantly enhanced apoptosis under hypoxia possibly by preventing the hypoxia mediated decrease in Bak expression without altering Bax expression. Conclusion: these results are a clear demonstration that hypoxia has a direct protective effect on apoptotic cell death. Moreover, molecular profiling points to putative pathways responsible for tumor growth in challenging environmental conditions and cancer cell resistance to chemotherapeutic agents

Earthworms Use Odor Cues to Locate and Feed on Microorganisms in Soil

Earthworms are key components of temperate soil ecosystems but key aspects of their ecology remain unexamined. Here we elucidate the role of olfactory cues in earthworm attraction to food sources and document specific chemical cues that attract Eisenia fetida to the soil fungi Geotrichum candidum. Fungi and other microorganisms are major sources of volatile emissions in soil ecosystems as well as primary food sources for earthworms, suggesting the likelihood that earthworms might profitably use olfactory cues to guide foraging behavior. Moreover, previous studies have documented earthworm movement toward microbial food sources. But, the specific olfactory cues responsible for earthworm attraction have not previously been identified. Using olfactometer assays combined with chemical analyses (GC-MS), we documented the attraction of E. fetida individuals to filtrate derived from G. candidum colonies and to two individual compounds tested in isolation: ethyl pentanoate and ethyl hexanoate. Attraction at a distance was observed when barriers prevented the worms from reaching the target stimuli, confirming the role of volatile cues. These findings enhance our understanding of the mechanisms underlying key trophic interactions in soil ecosystems and have potential implications for the extraction and collection of earthworms in vermiculture and other applied activities

Large expert-curated database for benchmarking document similarity detection in biomedical literature search

Document recommendation systems for locating relevant literature have mostly relied on methods developed a decade ago. This is largely due to the lack of a large offline gold-standard benchmark of relevant documents that cover a variety of research fields such that newly developed literature search techniques can be compared, improved and translated into practice. To overcome this bottleneck, we have established the RElevant LIterature SearcH consortium consisting of more than 1500 scientists from 84 countries, who have collectively annotated the relevance of over 180 000 PubMed-listed articles with regard to their respective seed (input) article/s. The majority of annotations were contributed by highly experienced, original authors of the seed articles. The collected data cover 76% of all unique PubMed Medical Subject Headings descriptors. No systematic biases were observed across different experience levels, research fields or time spent on annotations. More importantly, annotations of the same document pairs contributed by different scientists were highly concordant. We further show that the three representative baseline methods used to generate recommended articles for evaluation (Okapi Best Matching 25, Term Frequency-Inverse Document Frequency and PubMed Related Articles) had similar overall performances. Additionally, we found that these methods each tend to produce distinct collections of recommended articles, suggesting that a hybrid method may be required to completely capture all relevant articles. The established database server located at https://relishdb.ict.griffith.edu.au is freely available for the downloading of annotation data and the blind testing of new methods. We expect that this benchmark will be useful for stimulating the development of new powerful techniques for title and title/abstract-based search engines for relevant articles in biomedical research.Peer reviewe

Effet de l’hypoxie sur la résistance des cellules cancéreuses à l’étoposide : Etude de l’apoptose et de l’autophagie

L’hypoxie tumorale est une caractéristique du microenvironnement de la plupart des tumeurs solides. Elle correspond à la faible tension en oxygène observée au niveau de certaines zones de la tumeur et est la conséquence de la prolifération rapide des cellules tumorales et de la structure désordonnée de la tumeur et de sa vascularisation. L’hypoxie tumorale est souvent corrélée à un mauvais pronostic de guérison des patients car c’est un phénomène qui renforce l’agressivité des cellules cancéreuses et leur résistance à des thérapies anticancéreuses telles que la radiothérapie et la chimiothérapie. La résistance à la chimiothérapie est due à un effet direct de l’hypoxie, car les agents utilisés en chimiothérapie nécessitent souvent l’oxygène pour exercer leur cytotoxicité, et à un effet indirect de l’hypoxie qui induit une série d’adaptations au niveau de la cellule. L’hypoxie modifie le métabolisme de la cellule, stimule l’angiogenèse, l’érythropoïèse et régule la survie cellulaire en participant à la régulation de l’apoptose. La régulation de l’apoptose par l’hypoxie est complexe car en fonction de sa sévérité, l’hypoxie peut stimuler ou au contraire, inhiber l’apoptose. La compréhension de la régulation de l’apoptose par l’hypoxie est importante car l’apoptose participe à la sélection des cellules tumorales les plus agressives et conditionne l’efficacité des agents cytotoxiques utilisés au cours de la chimiothérapie. Ce travail a pour objectif d’étudier les mécanismes de régulation de l’apoptose par l’hypoxie. Dans un premier temps, nous avons comparé l’effet de l’hypoxie sur l’apoptose induite par l’étoposide, qui est un agent cytotoxique utilisé pour traiter certains types de cancers, au niveau de trois lignées cancéreuses humaines provenant de trois organes différents. Nous avons montré que l’effet de l’hypoxie sur l’apoptose induite par l’étoposide était dépendant de la lignée cellulaire car l’hypoxie stimule l’apoptose induite par l’étoposide dans les cellules MCF7 alors qu’elle inhibe l’apoptose induite par l’étoposide dans les cellules HepG2. Ceci montre que la régulation de l’apoptose par l’hypoxie est un mécanisme complexe, qui ne dépend pas uniquement de la sévérité de l’hypoxie, mais également d’autres paramètres tels que la lignée cellulaire. Dans la suite du travail, nous avons étudié comment l’hypoxie était capable de protéger les cellules HepG2 de l’apoptose induite par l’étoposide. Pour cela, nous avons évalué l’implication de plusieurs facteurs de transcription. Les résultats ont mis en évidence que l’hypoxie protège les cellules HepG2 de l’apoptose induite par l’étoposide en inhibant le facteur de transcription p53 et en activant le facteur de transcription c-jun. Nous avons également évalué l’implication de l’autophagie qui est un processus cellulaire impliqué dans le recyclage des protéines de longue durée de vie et des organites ainsi que dans la survie et la mort cellulaire. En condition d’hypoxie, l’autophagie peut être induite par un mécanisme impliquant la protéine BNIP3. Nous avons montré que, dans les cellules HepG2, l’autophagie était induite par l’étoposide mais pas par l’hypoxie. Cependant, l’autophagie induite par l’étoposide a des conséquences différentes en fonction de la tension en oxygène puisque les résultats ont montré que l’autophagie favorise l’apoptose induite par l’étoposide en normoxie mais pas en hypoxie. Enfin, nous avons montré que BNIP3 n’influençait pas l’autophagie mais participait à la protection contre l’apoptose induite par l’étoposide en hypoxie. En conclusion, les résultats obtenus montrent que la régulation de l’apoptose par l’hypoxie est un phénomène complexe qui est dépendant de plusieurs paramètres dont la lignée cellulaire. De plus, ces résultats indiquent que, lorsque l’hypoxie protège de l’apoptose, cette protection résulte de plusieurs adaptations mises en place par l’hypoxie. Ces résultats sont importants car ils améliorent notre compréhension des phénomènes de résistance observés chez les patients

Tumour Hypoxia Affects the Responsiveness of Cancer Cells to Chemotherapy and Promotes Cancer Progression

A solid tumour forms an organ-like structure that is comprised of cancer cells as well as stroma cells (fibroblasts, inflammatory cells) that are embedded in an extracellular matrix and are nourished by vascular network. However, tumoral microenvironment is heterogeneous due to the abnormal vasculature network and high proliferation rate of cancer cells. Because of these features, some regions are starved from oxygen, a phenomenon called hypoxia. Transient hypoxia is associated with inadequate blood flow while chronic hypoxia is the consequence of the increased oxygen diffusion distance due to tumour expansion. Both types of hypoxia are correlated with poor outcome for patients. Moreover, hypoxia also enhances chemoresistance of cancer cells. Firstly, the delivery of drugs in hypoxic area and cellular uptake of it are affected by hypoxia or associated acidity. Secondly, some chemotherapeutic drugs require oxygen to generate free radicals that contribute to cytotoxicity. Last, hypoxia induces cellular adaptations that compromise the effectiveness of chemotherapy. In response to nutrient deprivation due to hypoxia, the rate of proliferation of cancer cells decreases but chemotherapeutic drugs are more effective against proliferating cells. On the other hand, hypoxia induces adaptation by post-translational and transcriptional changes that promote cell survival and resistance to chemotherapy. Through these changes, hypoxia promotes angiogenesis, shift to glycolytic metabolism, expression of ABC transporters, cell survival by inducing the expression of genes encoding growth factors and the modulation of apoptotic process. The aim of this review is to provide a description of known hypoxia-induced mechanisms of chemoresistance at a cellular level

Etude de l'effet de composés bio-sourcés sur l'activation des cellules immunitaires en vue d'améliorer l'immunothérapie du mésothéliome pleural malin

Glycans are macromolecules (O-glycosidic and N-glycosidic linkage of monosacharides) which can be extracted from various agricultural products (i.e. cellulose, lignin, starch, insect). Preliminary data indicate that glycans can stimulate inflammation without initiating an anti-inflammatory feedback loop. Notwithstanding, inhibition of immune response by anti-inflammatory cytokines is a major cause of cancer therapy failure in human. The goal of the project is to study the ability of glycans to modulate the functions of macrophages and dendritic cells in order to stimulate anti-tumor immune response. The project aims to assess this hypothesis in a high incidence cancer: Malignant Pleural Mesothelioma. Glycan extracts will be isolated from a variety of agroforestry resources and especially broadleaved trees of the different systems tested on the platform AgricultureIsLife (zones 1-3 : hedges, tree lines, agrisilviculture, short rotation coppice) and modified by physical, chemical, enzymatic or microbiotic approaches