Calcium-binding protein S100A4 in health and disease

AbstractThe S100 proteins contain two EF-hand motifs and are of generally unknown function. One of these proteins, S100A4, is an intracellular calcium-binding protein that is present in normal rodent and human cells. In cultured rodent mammary cells, S100A4 is expressed at a higher level in some metastatic epithelial cells than in non-metastatic counterparts. Similarly, in human breast cell lines, S100A4 is present at a higher level in cultured cells from the more malignant, than in those from the more benign tumours. Gene transfer experiments have shown that rodent or human S100A4 is able to induce metastatic capability in otherwise non-metastatic breast tumour cells. Furthermore, expression of rodent S100A4 transgenes can induce metastasis of benign tumours arising in transgenic model systems. Possible mechanisms for the metastasis-inducing effect of S100A4 and the relevance of these observations to human cancer are discussed

Cathepsin Z as a novel potential biomarker for osteoporosis

Osteoporosis, one of the most prevalent chronic ageing-related bone diseases, often goes undetected until the first fragility fracture occurs, causing patient suffering and cost to health/social care services. Osteoporosis arises from imbalanced activity of osteoclasts and osteoblasts. Since these cell lineages produce the protease, cathepsin Z, the aim of this study was to investigate whether altered cathepsin Z mRNA levels are associated with osteoporosis in clinical samples. Cathepsin Z mRNA in human peripheral blood mononuclear cells was significantly differentially-expressed among non-osteoporotic controls, osteopenia and osteoporosis patients (p < 0.0001) and in female osteoporosis patients over the age of 50 years (P = 0.0016). Cathepsin Z mRNA level strongly correlated with low bone mineral density (BMD) (g/cm2), lumbar spine L2-L4 and femoral neck (T-scores) (P = 0.0149, 0.0002 and 0.0139, respectively). Importantly, cathepsin Z mRNA was significantly associated with fragility fracture in osteoporosis patients (P = 0.0018). The levels of cathepsin Z mRNA were not significantly higher in patients with chronic inflammatory disorders in these two groups compared to those without (P = 0.774 and 0.666, respectively). ROC analysis showed that cathepsin Z mRNA has strong diagnostic value for osteoporosis and osteoporotic fracture. The results show for the first time that cathepsin Z could be a future diagnostic biomarker for osteoporosis including female osteoporosis patients over the age of 50 years

Altered levels of mRNAs for calcium binding / associated proteins, Annexin A1, S100A4 and TMEM64 in peripheral blood mononuclear cells are associated with osteoporosis

Background. Osteoporosis is the most common metabolic bone disease in the world. Since osteoporosis is clinically symptomless until the first fracture occurs, early diagnosis is critical. Calcium, along with calcium-binding and calcium-associated proteins, plays an important role in homeostasis, maintaining healthy bone metabolism. This study is aimed at investigating the level of calcium-binding/associated proteins, annexin A1, S100A4, and TMEM64, in peripheral blood mononuclear cells associated with osteoporosis and its clinical significance. Methods. The levels of mRNAs of annexin A1, S100A4, and TMEM64 in human peripheral blood mononuclear cells were evaluated among 48 osteopenia and 23 osteoporosis patients compared to 17 nonosteoporotic controls. Total RNAs were isolated from clinical samples, and quantitation of mRNA levels was performed using real-time quantitative PCR. Results. The levels of mRNAs for calcium-binding proteins, annexin A1 and S100A4, and calcium-associated protein, TMEM64, in human peripheral blood mononuclear cells were significantly reduced in osteopenia and osteoporosis patients compared with nonosteoporotic controls (one-way ANOVA, P < 0.0001, P = 0.039 and P = 0.0195 respectively). Annexin A1 and TMEM64 mRNAs were also significantly reduced in female osteoporosis patients over the age of 50 years compared to nonosteoporotic controls (one-way ANOVA, P = 0.004 and P = 0.0037, respectively). ROC analysis showed that the reduction in the level of mRNA for annexin A1, S100A4, or TMEM64 in the patients’ peripheral blood mononuclear cells has a good diagnostic value for osteoporosis. Conclusions. The results show for the first time that calcium-binding/associated proteins, annexin A1 and TMEM64, could be future diagnostic biomarkers for osteoporosis

Assessing estrogen-induced proliferative response in an endometrial cancer cell line using a universally applicable methodological guide

Objective: Translational endometrial cancer (EC) research benefits from an in vitro experimental approach using EC cell lines. We demonstrated the steps that are required to examine estrogen induced proliferative response, a simple yet important research question pertinent to EC and devised a pragmatic methodological workflow for utilising EC cell lines in experimental models. Methods/materials: Comprehensive review of all commercially available EC cell lines was carried out, and Ishikawa cell line was selected to study the oestrogen responsiveness with HEC1A, RL95-2 and MFE280 cell lines as comparators where appropriate, examining relevant differential molecular (steroid receptors) and functional (phenotype, anchorage-independent growth, hormone responsiveness, migration, invasion and chemosensitivity) characteristics in 2D and 3D cultures in vitro using immunocytochemistry, immunofluorescence, qPCR and western blotting. In vivo tumour, formation and chemosensitivity were also assessed in a chick chorioallantoic membrane (CAM) model. Results: Short Tandem Repeat (STR) analysis authenticated the purchased cell lines while gifted cells deviated significantly from the published profile. We demonstrate the importance of prior assessment of the suitability of each cell line for the chosen in vitro experimental technique. Prior establishment of baseline, non-enriched conditions was required to induce a proliferative response to estrogen. The CAM model was a suitable in vivo multi-cellular animal model for EC, for producing rapid and reproducible data. Conclusions: We have developed a methodological guide for EC researchers when using endometrial cell lines to answer important translational research questions (exemplified by estrogen responsive cell proliferation), to facilitate robust data, while saving time and resources

The role of the C-terminal lysine of S100P in S100P-induced cell migration and metastasis

S100P protein is a potent inducer of metastasis in a model system, and its presence in cancer cells of patients is strongly associated with their reduced survival times. A well-established Furth Wistar rat metastasis model system, methods for measuring cell migration, and specific inhibitors were used to study pathways of motility-driven metastasis. Cells expressing C-terminal mutant S100P proteins display markedly-reduced S100P-driven metastasis in vivo and cell migration in vitro. These cells fail to display the low focal adhesion numbers observed in cells expressing wild-type S100P, and the mutant S100P proteins exhibit reduced biochemical interaction with non-muscle myosin heavy chain isoform IIA in vitro. Extracellular inhibitors of the S100P-dependent plasminogen activation pathway reduce, but only in part, wild-type S100P-dependent cell migration; they are without effect on S100P-negative cells or cells expressing C-terminal mutant S100P proteins and have no effect on the numbers of focal adhesions. Recombinant wild-type S100P protein, added extracellularly to S100P-negative cells, stimulates cell migration, which is abolished by these inhibitors. The results identify at least two S100P-dependent pathways of migration, one cell surface and the other intracellularly-linked, and identify its C-terminal lysine as a target for inhibiting multiple migration-promoting activities of S100P protein and S100P-driven metastasis

Comparison Of The Genesis Solar Wind Regime Algorithm Results With Solar Wind Composition Observed By ACE

Launched on 8 August 2001, the NASA Genesis mission is now collecting samples of the solar wind in various materials, and will return those samples to Earth in 2004 for analysis. A primary science goal of Genesis is the determination of the isotopic and elemental composition of the solar atmosphere from the solar wind material returned. In particular, Genesis will provide measurements of those species that are not provided by solar and in situ observations. We know from in situ measurements that the solar wind exhibits compositional variations across different types of solar wind flows. Therefore, Genesis exposes different collectors to solar wind originating from three flow types: coronal hole, coronal mass ejection (CME), and interstream flows. Flow types are identified using in situ measurements of solar wind protons, alphas, and electrons from electrostatic analyzers carried by Genesis. The flow regime selection algorithm and subsequent collector deployment on Genesis act autonomously. We present an assessment of composition variations of O, He, and Mg ions observed by ACE/SWICS concurrent with Genesis observations, and compare these to the Genesis algorithm decisions. Not only does this serve as a test of the algorithm, the compilation of composition vs. regime will be important for comparison to the abundances determined from sample analysis at the end of the mission. © 2003 American Institute of PhysicsPeer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87657/2/632_1.pd