Progress in epithelial-mesenchymal transition research

The field of research of epithelial-mesenchymal transitions, EMT, and its reverse, mesenchymal-epithelial transitions, MET, has expanded very rapidly indeed from its beginnings, heralded by Professor Betty Hay in the 1970s and 1980s. This expansion has involved the realisation that the EMT was not just an interesting phenomenon of early developmental morphogenetic cell behaviour, but bore remarkable resemblance to clinically crucial pathological events in cancer invasion. Not surprisingly, this discipline soon became numerically dominant in the EMT publication field. Simultaneously, the EMT concept has been extended to normal physiological wound healing. Exploration revealed that these resemblances were more than skin deep: the same sets of growth factors, receptors, transcription factors, epigenetic marks and signalling pathways turned up repeatedly in EMTs and METs in a variety of contexts, both pathological and normal. This molecular genetic research in turn uncovered similarities of the EMT signature to that of fibrosis, a set of diseases which is of enormous clinical importance, rivalling that of cancer. Most recently, and more surprisingly, the EMT signature has shown considerable similarity to that found in stem cell and cancer stem cell biology.

Assessment of gene expression of intracellular calcium channels, pumps and exchangers with epidermal growth factor-induced epithelial-mesenchymal transition in a breast cancer cell line

Background Epithelial-mesenchymal transition (EMT) is a process implicated in cancer metastasis that involves the conversion of epithelial cells to a more mesenchymal and invasive cell phenotype. In breast cancer cells EMT is associated with altered store-operated calcium influx and changes in calcium signalling mediated by activation of cell surface purinergic receptors. In this study, we investigated whether MDA-MB-468 breast cancer cells induced to undergo EMT exhibit changes in mRNA levels of calcium channels, pumps and exchangers located on intracellular calcium storing organelles, including the Golgi, mitochondria and endoplasmic reticulum (ER). Methods Epidermal growth factor (EGF) was used to induce EMT in MDA-MB-468 breast cancer cells. Serum-deprived cells were treated with EGF (50 ng/mL) for 12 h and gene expression was assessed using quantitative RT-PCR. Results and conclusions These data reveal no significant alterations in mRNA levels of the Golgi calcium pump secretory pathway calcium ATPases (SPCA1 and SPCA2), or the mitochondrial calcium uniporter (MCU) or Na+/Ca2+ exchanger (NCLX). However, EGF-induced EMT was associated with significant alterations in mRNA levels of specific ER calcium channels and pumps, including (sarco)-endoplasmic reticulum calcium ATPases (SERCAs), and inositol 1,4,5-trisphosphate receptor (IP3R) and ryanodine receptor (RYR) calcium channel isoforms. The most prominent change in gene expression between the epithelial and mesenchymal-like states was RYR2, which was enriched 45-fold in EGF-treated MDA-MB-468 cells. These findings indicate that EGF-induced EMT in breast cancer cells may be associated with major alterations in ER calcium homeostasis.

Induction of epithelial-mesenchymal transition (EMT) in breast cancer cells is calcium signal dependent

Signals from the tumor microenvironment trigger cancer cells to adopt an invasive phenotype through epithelial-mesenchymal transition (EMT). Relatively little is known regarding key signal transduction pathways that serve as cytosolic bridges between cell surface receptors and nuclear transcription factors to induce EMT. A better understanding of these early EMT events may identify potential targets for the control of metastasis. One rapid intracellular signaling pathway that has not yet been explored during EMT induction is calcium. Here we show that stimuli used to induce EMT produce a transient increase in cytosolic calcium levels in human breast cancer cells. Attenuation of the calcium signal by intracellular calcium chelation significantly reduced epidermal growth factor (EGF)- and hypoxia-induced EMT. Intracellular calcium chelation also inhibited EGF-induced activation of signal transducer and activator of transcription 3 (STAT3), while preserving other signal transduction pathways such as Akt and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation. To identify calcium-permeable channels that may regulate EMT induction in breast cancer cells, we performed a targeted siRNA-based screen. We found that transient receptor potential-melastatin-like 7 (TRPM7) channel expression regulated EGF-induced STAT3 phosphorylation and expression of the EMT marker vimentin. Although intracellular calcium chelation almost completely blocked the induction of many EMT markers, including vimentin, Twist and N-cadherin, the effect of TRPM7 silencing was specific for vimentin protein expression and STAT3 phosphorylation. These results indicate that TRPM7 is a partial regulator of EMT in breast cancer cells, and that other calcium-permeable ion channels are also involved in calcium-dependent EMT induction. In summary, this work establishes an important role for the intracellular calcium signal in the induction of EMT in human breast cancer cells. Manipulation of calcium-signaling pathways controlling EMT induction in cancer cells may therefore be an important therapeutic strategy for preventing metastases.

Exploring young student's functional thinking (Exploración del pensamiento funcional de estudiantes jóvenes)

The Early Years Generalizing Project (EYGP) involves Australian years 1 to 4 (age 5 to 9) students and investigates how they grasp and express generalizations. This paper focuses on data collected from 6 Year 1 students in an exploratory study within a clinical interview setting that required students to identify function rules. Preliminary findings suggest that the use of gestures (both by students and interviewers), self-talk (by students), and concrete acting out, assisted students to reach generalizations and to begin to express these generalities. It also appears that as students became aware of the structure, their use of gestures and selftalk tended to decrease.

ORIGO Stepping Stones Grade K Student Journal

A mathematics curriculum for the Common Core Curriculum at the Kindergarten level for the USA

Teacher organizational citizenship behaviours and job efficacy : implications for student quality of school life

The present study investigated the impact of teachers' organizational citizenship behaviours (OCBs) on student quality of school life (SQSL) via the indirect effect of job efficacy. A measure of teacher OCBs was developed, tapping one dimension of individual-focused OCB (OCBI – student-directed behaviour) and two dimensions of organization-focused OCB (OCBO – civic virtue and professional development). In line with previous research suggesting that OCBs may enhance job efficacy, as well as studies demonstrating the positive effects of teacher efficacy on student outcomes, we expected an indirect relationship between teachers OCBs and SQSL via teachers' job efficacy. Hypotheses were tested in a multi-level design in which 170 teachers and their students (N=3,057) completed questionnaires. A significant proportion of variance in SQSL was attributable to classroom factors. Analyses revealed that the civic virtue and professional development behaviours of teachers were positively related to their job efficacy. The job efficacy of teachers also had a positive impact on all five indicators of SQSL. In regards to professional development, job efficacy acted as an indirect variable in the prediction of four student outcomes (i.e., general satisfaction, student–teacher relations, achievement, and opportunity) and fully mediated the direct negative effect on psychological distress.

Induction of epithelial to mesenchymal transition in PMC42-LA human breast carcinoma cells by carcinoma-associated fibroblast secreted factors

Background Breast carcinoma is accompanied by changes in the acellular and cellular components of the microenvironment, the latter typified by a switch from fibroblasts to myofibroblasts. Methods: We utilised conditioned media cultures, Western blot analysis and immunocytochemistry to investigate the differential effects of normal mammary fibroblasts (NMFs) and mammary cancer-associated fibroblasts (CAFs) on the phenotype and behaviour of PMC42-LA breast cancer cells. NMFs were obtained from a mammary gland at reduction mammoplasty, and CAFs from a mammary carcinoma after resection. Results We found greater expression of myofibroblastic markers in CAFs than in NMFs. Medium from both CAFs and NMFs induced novel expression of α-smooth muscle actin and cytokeratin-14 in PMC42-LA organoids. However, although conditioned media from NMFs resulted in distribution of vimentin-positive cells to the periphery of PMC42-LA organoids, this was not seen with CAF-conditioned medium. Upregulation of vimentin was accompanied by a mis-localization of E-cadherin, suggesting a loss of adhesive function. This was confirmed by visualizing the change in active β-catenin, localized to the cell junctions in control cells/ cells in NMF-conditioned medium, to inactive β-catenin, localized to nuclei and cytoplasm in cells in CAF-conditioned medium. Conclusion We found no significant difference between the influences of NMFs and CAFs on PMC42-LA cell proliferation, viability, or apoptosis; significantly, we demonstrated a role for CAFs, but not for NMFs, in increasing the migratory ability of PMC42-LA cells. By concentrating NMF-conditioned media, we demonstrated the presence of factor(s) that induce epithelial-mesenchymal transition in NMF-conditioned media that are present at higher levels in CAF-conditioned media. Our in vitro results are consistent with observations in vivo showing that alterations in stroma influence the phenotype and behaviour of surrounding cells and provide evidence for a role for CAFs in stimulating cancer progression via an epithelial-mesenchymal transition. These findings have implications for our understanding of the roles of signalling between epithelial and stromal cells in the development and progression of mammary carcinoma.

Epithelial to mesenchymal transition and breast cancer

Epithelial-mesenchymal plasticity in breast carcinoma encompasses the phenotypic spectrum whereby epithelial carcinoma cells within a primary tumor acquire mesenchymal features and re-epithelialize to form a cohesive secondary mass at a metastatic site. Such plasticity has implications in progression of breast carcinoma to metastasis, and will likely influence response to therapy. The transcriptional and epigenetic regulation of molecular and cellular processes that underlie breast cancer and result in characteristic changes in cell behavior can be monitored using an increasing array of marker proteins. Amongst these markers exists the potential for emergent prognostic, predictive and therapeutic targeting.

Peer assisted learning and generation Y : a case study

In recent years Australian Law Schools have implemented various forms of peer assisted learning or mentoring, including career mentoring by former students of final year students and orientation mentoring or tutoring by later year students of incoming first year students. The focus of these programs therefore is on the transition into or out of law school. There is not always as great an emphasis however, as part of this transition, on the use of law students belonging to the same unit cohort as a learning resource for each other within their degree. This is despite the claimed preference of Generation Y students for collaborative learning environments, authentic learning experiences and the development of marketable workplace skills. In the workplace, be it professional legal practice or otherwise, colleagues rely heavily on each other for information, support and guidance. In the undergraduate law degree at the Queensland University of Technology (‘QUT’) the Torts Student Peer Mentor Program aims to supplement a student’s understanding of the substantive law of torts with the development of life-long skills. As such it has the primary objective, albeit through discussion facilitated by more senior students, of encouraging first year students to develop for themselves the skills they need to be successful both as law students and as legal practitioners. Examples of such skills include those relevant to: preparation for assessment tasks; group work; problem solving, cognition and critical thinking; independent learning; and communication. Significantly, in this way, not only do the mentees benefit from involvement in the program, but the peer mentors, or program facilitators, themselves also benefit from their participation in the real world learning environment the program provides. This paper outlines the development and implementation of the above program, the pedagogy which influenced it, and its impact on student learning experiences

Effect of crack depth and specimen width on fracture toughness of a carbon steel in the ductile-brittle transition region

The effects of crack depth (a/W) and specimen width W on the fracture toughness and ductile±brittle transition have been investigated using three-point bend specimens. Finite element analysis is employed to obtain the stress-strain fields ahead of the crack tip. The results show that both normalized crack depth (a/W) and specimen width (W) affect the fracture toughness and ductile±brittle fracture transition. The measured crack tip opening displacement decreases and ductile±brittle transition occurs with increasing crack depth (a/W) from 0.1 to 0.2 and 0.3. At a fixed a/W (0.2 or 0.3), all specimens fail by cleavage prior to ductile tearing when specimen width W increases from 25 to 40 and 50 mm. The lower bound fracture toughness is not sensitive to crack depth and specimen width. Finite element analysis shows that the opening stress in the remaining ligament is elevated with increasing crack depth or specimen width due to the increase of in-plane constraint. The average local cleavage stress is dependent on both crack depth and specimen width but its lower bound value is not sensitive to constraint level. No fixed distance can be found from the cleavage initiation site to the crack tip and this distance increases gradually with decreasing inplane constraint.