Envisioned System Research Articles

Pricing New Medicines in the UK: ‘Price is what you Pay, Value is what you Get'?

Warren Buffett's aphorism neatly captures the ambition underlying proposals which have been put forward for reform of regulation of pharmaceutical pricing in the UK's National Health Service. Through a critical evaluation of the envisioned system of 'value-based pricing', this article explores differing understandings of the value which a new medicine delivers to patients and society. It further considers the possible consequences for regulatory legitimacy of the adoption of the more pluralistic approach to value which the proposals embody.

Modulation of lipid metabolism with the overexpression of NPC1L1 in mouse liver

Niemann-Pick C1-like 1 protein (NPC1L1), a transporter crucial in intestinal cholesterol absorption, is expressed in human liver but not in murine liver. To elucidate the role of hepatic NPC1L1 on lipid metabolism, we overexpressed NPC1L1 in murine liver utilizing adenovirus-mediated gene transfer. C57BL/6 mice, fed on normal chow with or without ezetimibe, were injected with NPC1L1 adenovirus (L1-mice) or control virus (Null-mice), and lipid analyses were performed five days after the injection. The plasma cholesterol levels increased in L1-mice, and FPLC analyses revealed increased cholesterol contents in large HDL lipoprotein fractions. These fractions, which showed α-mobility on agarose electrophoresis, were rich in apoE and free cholesterol. These lipoprotein changes were partially inhibited by ezetimibe treatment and were not observed in apoE-deficient mice. In addition, plasma and VLDL triglyceride (TG) levels decreased in L1-mice. The expression of microsomal triglyceride transfer protein (MTP) was markedly decreased in L1-mice, accompanied by the reduced protein levels of forkhead box protein O1 (FoxO1). These changes were not observed in mice with increased hepatic de novo cholesterol synthesis. These data demonstrate that cholesterol absorbed through NPC1L1 plays a distinct role in cellular and plasma lipid metabolism, such as the appearance of apoE-rich lipoproteins and the diminished VLDL-TG secretion.

Perivascular Epithelioid Cell Tumors (PEComas) Harboring TFE3 Gene Rearrangements Lack the TSC2 Alterations Characteristic of Conventional PEComas

Perivascular Epithelioid Cell Tumors (PEComas) Harboring TFE3 Gene Rearrangements Lack the TSC2 Alterations Characteristic of Conventional PEComas

CD34 expression in human hair follicles and tricholemmoma: a comprehensive study

There has recently been controversy regarding whether clone My10 is superior to clone QBEND-10 for labeling cells of tricholemmal lineage. Moreover, there have been no previous reports on the CD34 expression in human vellus hair follicles. We performed a comprehensive study of the CD34 expression in human terminal and vellus hair follicles and in 10 tricholemmomas using both the QBEND-10 and the My10 clones. We also performed two different procedures of immunostaining, which included the using of the standard avidin-biotin-peroxidase (ABC) complex system and the Envision system. The most sensitive marker of CD34 for normal human hair follicles and tricholemmomas is QBEND-10 using the ABC system. The degree and strength of the CD34 positive staining mainly depended on the method being used (whether it was the ABC system or the Envision system) rather than the clone. CD34 staining was rarely (20-30%) seen in the anagen and catagen vellus hair follicles, and could only be seen by the QBEND-10 clone using the ABC system. CD34 expression in the tricholemmomas represented either a diffuse or peripheral pattern. CD34 may not be a tricholemmal lineage-specific antigen, but may be related to certain functions of the cells.

Isolation and Characterization of Progenitor-Like Cells from Human Renal Proximal Tubules

The tubules of the kidney display a remarkable capacity for self-renewal on damage. Whether this regeneration is mediated by dedifferentiating surviving cells or, as recently suggested, by stem cells has not been unequivocally settled. Herein, we demonstrate that aldehyde dehydrogenase (ALDH) activity may be used for isolation of cells with progenitor characteristics from adult human renal cortical tissue. Gene expression profiling of the isolated ALDH(high) and ALDH(low) cell fractions followed by immunohistochemical interrogation of renal tissues enabled us to delineate a tentative progenitor cell population scattered through the proximal tubules (PTs). These cells expressed CD24 and CD133, previously described markers for renal progenitors of Bowman's capsule. Furthermore, we show that the PT cells, and the glomerular progenitors, are positive for KRT7, KRT19, BCL2, and vimentin. In addition, tubular epithelium regenerating on acute tubular necrosis displayed long stretches of CD133(+)/VIM(+) cells, further substantiating that these cells may represent a progenitor cell population. Furthermore, a potential association of these progenitor cells with papillary renal cell carcinoma was discovered. Taken together, our data demonstrate the presence of a previously unappreciated subset of the PT cells that may be endowed with a more robust phenotype, allowing increased resistance to acute renal injury, enabling rapid repopulation of the tubules.

Strong BCL2 expression in Burkitt lymphoma is not uncommon in adults

The distinction between Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) is not always easy, so much so that the WHO 2008 Blue Book has incorporated a provisional category of "B-cell lymphoma, unclassifiable with features intermediate between DLBCL and BL." One of the immunohistochemical (IHC) markers used at times to differentiate between the two is IHC expression of BCL2, which was initially believed to be consistently negative in BL. Later it was recognized that weak BCL2 expression is still compatible with the diagnosis of BL. To verify or otherwise this current view this study was undertaken. We retrieved 39 confirmed cases of BL, in both children and adults. All these cases had typical morphology, IHC profile, and Mib-1 index that are typical of BL. All these cases were then stained with a monoclonal antibody against BCL2 oncoprotein, using 2-step Envision system. Out of 39 cases, 31 cases (79.4%) were completely negative for BCL2, whereas 5 (12.8%) were weak focal positive. However, another 4 (10.2%) cases did show strong diffuse cytoplasmic staining for BCL2. Fluorescent in-Situ hybridization (FISH) for t(14:18) was optimally done on 6 out of 9 cases. All these 4 cases were from adults with 3 out of 4 arising in the parotid region. Two out of 4 cases also showed t(8:14) on FISH. It was concluded that contrary to the common belief, strong BCL2 IHC expression is possible in typical BL in adults and cannot be absolutely relied upon to distinguish between BL and DLBCL.

Increased Number of B-Cells In the Red Pulp of the Spleen In ITP

AbstractAbstract 1447 Background.Platelets are targeted by autoantibodies and destroyed in the reticuloendothelial system, e.g. the spleen, in patients with chronic immune thrombocytopenia (ITP). However, other pathophysiologic mechanisms such as platelet destruction by cytotoxic T-cells and defective bone marrow production of platelets also contribute to the thrombocytopenia in ITP. Splenectomy normalizes the platelet count in around 70% of chronic ITP patients, however, precious little is known about the spleen micromorphology in this disease. We examined splenic pathology in patients with ITP, focusing on the intrasplenic distribution of lymphocyte subsets. Patients and Methods.The paraffin-embedded block containing representative material of the splenic tissue from 29 splenectomized chronic ITP patients (15 females and 14 males, mean age 45±21 (SD) years) and 11 controls, splenectomized because of traumatic splenic rupture (2 females and 9 males, mean age 48±19 (SD) years), was used to produce serial sections with a thickness of 5 μm. These sections were stained with conventional haematoxylin-eosin, and immunostained to identify CD3, CD4, CD8, CD20 and CD68 using the Dako Envision System in a Techmate Horizon Autostainer (Glostrup, Denmark). Using micromorphometry the lymphocyte subsets were enumerated in the red and white pulp of these spleens. Results.All ITP patients were preoperatively optimized, using corticosteroids and/or IVIg, and the immediate pre-splenectomy platelet count ranged 5–357 × 109/l.Significantly more T-cells than B-cells were seen in the red pulp, both in ITP and control spleens (p<0.01). Conversely, more B-cells than T-cells were observed in the white pulp. There was no difference in the number of T-cells in the red and white pulp, between ITP patients and controls. However, the number of B-cells was increased in the red pulp of patients with ITP compared with the controls. Furthermore, numerous B-cells and CD8+ (cytotoxic) T-cells were located in the splenic cords, lining the sinusoids in the red pulp, a microenvironment where they readily can interact with the other blood cells, e.g. platelets. Conclusion.Chronic ITP is associated with an increased number of CD20+ B-cells in the red pulp of the spleen. The red pulp of the spleen also provides a microenvironment where close interaction between platelets, CD3+, CD4+, and CD8+ T-cells and B-cells can take place. Trafficking of regulatory and effector cells into this microenvironment might be a potential therapeutic target. Disclosures:No relevant conflicts of interest to declare.

Mixed Lineage Kinase-3 Stabilizes and Functionally Cooperates with TRIBBLES-3 to Compromise Mitochondrial Integrity in Cytokine-induced Death of Pancreatic Beta Cells

Mixed lineage kinases (MLKs) have been implicated in cytokine signaling as well as in cell death pathways. Our studies show that MLK3 is activated in leukocyte-infiltrated islets of non-obese diabetic mice and that MLK3 activation compromises mitochondrial integrity and induces apoptosis of beta cells. Using an ex vivo model of islet-splenocyte co-culture, we show that MLK3 mediates its effects via the pseudokinase TRB3, a mammalian homolog of Drosophila Tribbles. TRB3 expression strongly coincided with conformational change and mitochondrial translocation of BAX. Mechanistically, MLK3 directly interacted with and stabilized TRB3, resulting in inhibition of Akt, a strong suppressor of BAX translocation and mitochondrial membrane permeabilization. Accordingly, attenuation of MLK3 or TRB3 expression each prevented cytokine-induced BAX conformational change and attenuated the progression to apoptosis. We conclude that MLKs compromise mitochondrial integrity and suppress cellular survival mechanisms via TRB3-dependent inhibition of Akt.

EphA2 Mutation in Lung Squamous Cell Carcinoma Promotes Increased Cell Survival, Cell Invasion, Focal Adhesions, and Mammalian Target of Rapamycin Activation

Non-small cell lung cancer (NSCLC) has a poor prognosis and improved therapies are needed. Expression of EphA2 is increased in NSCLC metastases. In this study, we investigated EphA2 mutations in NSCLC and examined molecular pathways involved in NSCLC. Tumor and cell line DNA was sequenced. One EphA2 mutation was modeled by expression in BEAS2B cells, and functional and biochemical studies were conducted. A G391R mutation was detected in H2170 and 2/28 squamous cell carcinoma patient samples. EphA2 G391R caused constitutive activation of EphA2 with increased phosphorylation of Src, cortactin, and p130(Cas). Wild-type (WT) and G391R cells had 20 and 40% increased invasiveness; this was attenuated with knockdown of Src, cortactin, or p130(Cas). WT and G391R cells demonstrated a 70% increase in focal adhesion area. Mammalian target of rapamycin (mTOR) phosphorylation was increased in G391R cells with increased survival (55%) compared with WT (30%) and had increased sensitivity to rapamycin. A recurrent EphA2 mutation is present in lung squamous cell carcinoma and increases tumor invasion and survival through activation of focal adhesions and actin cytoskeletal regulatory proteins as well as mTOR. Further study of EphA2 as a therapeutic target is warranted.

Immunohistochemical detection of Ki-67 is not associated with tumor-infiltrating macrophages and cyclooxygenase-2 in oral squamous cell carcinoma

An inflammatory component consisting of cells and chemical mediators may influence the proliferation and dissemination of the oral squamous cell carcinoma (OSCC). In the present study, we evaluated the possible relationship between Ki-67, tumor-associated macrophages (TAMs), and COX-2 in OSCCs. In addition, the immunodetection of these proteins was associated with different histological grades of malignancy, including invasive and in situ tumors. Twenty-seven OSCC cases were examined by light microscopy using criteria adopted WHO, and immunohistochemistry for Ki-67, CD68, and COX-2 using EnVision System in invasive and in situ lesions. Immunohistochemical detection of these proteins was assessed and scored for COX-2, and results were compared with their histological grades of malignancy. A correlation between Ki-67, COX-2, and CD68 was not found. Histological grade of malignancy (HDM) was associated with the Ki-67 immunostaining (P = 0.00), but this was not observed regarding both CD68 (P = 0.51) and COX-2 (P = 0.89). Furthermore, there was a COX-2 overexpression in 62.96% of the sample, and a high density of TAMs in both OSCCs and in situ carcinomas. Imunolabeling for Ki-67 was directly correlated with less-differentiated tumors, suggesting that this marker may contribute to understand the biological behavior of OSCC, and help to distinguish risk groups of OSCC. Furthermore, the lack of correlation between Ki-67, COX-2, and CD68 indicates that the latter two markers may play a pivotal role in oral carcinogenesis. However, further studies are needed to clarify their contribution for cell proliferation and tumor differentiation.

Multicriteria Decision Analysis Method for Evaluation of Software Architectures

architectures is a critical aspect in the design and development of software. Architecture of software is a collection of design decisions that are expensive to change. Software architectures are generally designed with particular functional and non-functional requirements. Organizations often need to choose software architecture for future development from several competing candidate architectures. The various stakeholders quality requirements need to be considered collectively to describe the quality requirements of the envisioned system and therefore build the basis for the comparison and selection criteria. Given the impact that software architecture has on a projects success, the need to choose the right architecture assumes significance. In this paper, a new architecture selection method based on multicriteria decision analysis have been developed and validated using a suitable case study.

A decision-support system for improving road transportation performance in Gauteng, South Africa: Developing a data acquisitioning system

This research describes how the systems approach can be applied to implement a holistic transportation decision-support system, which will measure transportation performance at the highest level and enable decision-making with respect to possible improvements. The research starts with the creation of a framework of the process to be followed. A framework of the envisioned system with existing and new elements is then created. Furthermore, a framework for management reports is proposed. Against this background, the deliverables for a project to implement the proposed data-acquisitioning system are defined. First order indications are that such a project is feasible and affordable, and can be completed to such an extent that most of the advantages are obtainable within two years.

Analysis of prognostic factors of retinoblastoma

To study the prognostic factors of retinoblastoma (RB). Prospective case series study.With Envision System, surgical specimens from 31 RB cases were immunostained by using anti-NSE, GFAP and S-100 antibodies. Number of NSE positive cells was counted at 1000 tumor cells in each slice. The morphology of GFAP and S-100 stained positive cells were analyzed. All cases were followed-up for 5 to 17 years. Cox regression pattern was adopted to analyze the relationship between sex, age, stage, differentiation, number of NSE positive cells, the sizes of GFAP and S-100 positive cells, therapies and prognosis. (1) Almost all cases were stained positively by anti-NSE antibodies. Positive rates of GFAP and S-100 expression specimens were 61.29% (19/31) and 58.06% (18/31), respectively. (2) The number of NSE positive cells was correlated with a better prognosis (P = 0.0361). The sizes of GFAP and S-100 stained area were also related with a better progress (P = 0.0430, 0.0477). In patients with a greater number of NSE stained cells (averaged 324/1000), the survival time was >/= 5 years. In patients with a less number of NSE stained cells (averaged 182/1000), the survival time was less than 5 years. (3) There was a significant correlation between the number of NSE stained cells and the results for GFAP and S-100 staining (r = 0.547, 0.581), respectively. The number of NSE positive cells is correlated with a better prognosis of RB. The expression of GFAP and S-100 protein are also correlated with a better prognosis of RB. The number of NSE positive cells and the expression of GFAP and S-100 protein may be regarded as the standards in judging the degree of differentiation of RB, and may be used as one of criteria for the estimation of the prognosis.

Immunohistochemistry for human telomerase reverse transcriptase catalytic subunit (hTERT): a study of 143 benign and malignant soft tissue and bone tumours

Only a small number of mesenchymal tumours have been previously evaluated for hTERT expression. We hypothesised that hTERT expression would be frequently present in sarcomas, but not in benign tumours. Sections from 143 bone/soft tissue tumours were immunostained for hTERT (44F12, 1:20; Novocastra) using steam heat-induced epitope retrieval and the Dako Envision system. Normal lymphocytes served as positive internal controls. Positive cases showed 'speckled' nuclear/nucleolar staining in >10% of cells, in accordance with previous studies. Fisher's exact test was used for statistical analysis. The result was checked against the G test and Chi-squared test with Yates correction, and was concordant. hTERT expression was seen in nine of 71 (13%) benign and 33 of 72 (46%) malignant tumours (p > or = 0.001). hTERT was positive in >50% of osteosarcomas, malignant peripheral nerve sheath tumours, liposarcomas, and angiosarcomas. All positive sarcomas were high grade (FNCLCC grade II or III). Among benign tumours, only schwannomas and chondromas were positive in >30% of cases. Positive normal tissues included lymphocytes, multinucleated histiocytes, keratinocytes, adnexal glands, hepatocytes and basal cells of respiratory mucosa. hTERT expression is significantly more common in sarcomas as compared with benign tumours. Restriction of hTERT expression to high grade lesions, and its absence in low grade sarcomas (including the well-differentiated component of dedifferentiated liposarcoma), suggests that telomerase activation is a late event in sarcoma progression. Importantly, however, hTERT can be expressed in some benign tumours, notably schwannoma and chondroma, and it is doubtful that hTERT expression alone will allow the discrimination of benign from malignant soft tissue/bone tumours. Importantly, hTERT expression is often only focal and its identification may require careful evaluation of an entire section, suggesting that this technique is best applied to whole sections, rather than tissue microarrays.

Global Levels of Histone Modifications Predict Prognosis in Different Cancers

Cancer cells exhibit alterations in histone modification patterns at individual genes and globally at the level of single nuclei in individual cells. We demonstrated previously that lower global/cellular levels of histone H3 lysine 4 dimethylation (H3K4me2) and H3K18 acetylation (ac) predict a higher risk of prostate cancer recurrence. Here we show that the cellular levels of both H3K4me2 and H3K18ac also predict clinical outcome in both lung and kidney cancer patients, with lower levels predicting significantly poorer survival probabilities in both cancer groups. We also show that lower cellular levels of H3K9me2, a modification associated with both gene activity and repression, is also prognostic of poorer outcome for individuals with either prostate or kidney cancers. The predictive power of these histone modifications was independent of tissue-specific clinicopathological variables, the proliferation marker Ki-67, or a p53 tumor suppressor mutation. Chromatin immunoprecipitation experiments indicated that the lower cellular levels of histone modifications in more aggressive cancer cell lines correlated with lower levels of modifications at DNA repetitive elements but not with gene promoters across the genome. Our results suggest that lower global levels of histone modifications are predictive of a more aggressive cancer phenotype, revealing a surprising commonality in prognostic epigenetic patterns of adenocarcinomas of different tissue origins.

Optimal modulation for known interference

We present a symbol-by-symbol approach to the problem of canceling known interference at the transmitter in a communication system. In the envisioned system, the modulator maps an information symbol (taken from a finite alphabet) and an interference symbol (from the complex field) onto a transmitted constellation point. Our scheme is based on joint optimization of a modulator and demodulator, subject to a constraint on the average transmit power. The demodulator picks the information symbol (as a function of the received symbol) that minimizes the average error probability. We emphasize that our focus is on transmission in a single (complex) dimension, and hence the proposed technique is a "modulation" rather than a "coding" scheme. We illustrate that the new scheme outperforms Tomlinson-Harashima precoding, which is a classical but suboptimal solution to the one-dimensional known-interference precoding problem. In our simulations, the new approach is able to perform close to the no-interference bound.

An update on privacy in ubiquitous computing

One of the editors of this special issue recently conducted an analysis of all major applications presented in the IEEE Pervasive Computing Magazine from 2003 to 2005. More than two-thirds of them had people as their main object of observation (rather than, say, ducks, bridges, or glaciers). In 90% of these human-focused applications, the observing person was not necessarily identical with the observed. Only about half of these applications allowed the people observed to even get feedback about (or insights into) their own behavior. In all other cases, third parties or machines were observing them without notice. These numbers illustrate why Ubicomp scholars regularly echo privacy as a key challenge for the adoption and ethical acceptability of smart environments. The unprecedented collection coverage, the invisibility of the collection process, the amount of data collected, and the envisioned system interconnectivity should motivate the community to consider ‘built-in’ privacy to a much larger extent than this may have been the case in earlier computing eras. It is for this reason that Personal and Ubiquitous Computing pays attention to the subject of privacy at regular intervals. The last special issue on privacy was published in May 2005 (Vol. 9, No. 3). The goal of this issue is to update the community with respect to the latest findings and developments surrounding privacy in Ubiquitous Computing. Unlike a normal call for papers, this special issue builds on the results of the 5th International Workshop on Ubicomp Privacy at the 9th International Conference on Ubiquitous Computing (UbiComp 2007) in Innsbruck in September 2007. The workshop brought together a diversity of research angles, grouped around five distinct themes: location privacy, RFID and sensor related privacy issues, consumer acceptance of RFID, and legal aspects of smart environments. This special issue presents the five original articles from the invited experts at the workshop (including ourselves). The first article by John Krumm offers a comprehensive but concise overview of location privacy: What is location privacy and how do people relate to it? How can location privacy be undermined? And what technical proposals exist to ensure ‘computational location privacy’? Krumm cites studies that show how people are generally not concerned about location privacy, and demonstrates how researchers have been able to link anonymized and obfuscated location tracks to the correct identity. While proposals exist to improve existing anonymization and obfuscation mechanisms, the article notes the still open issue of measuring the level of protection any such algorithms can offer. Norman Sadeh and his colleagues then illustrate the intricate usability issues inherent in providing location privacy in smart environments. An actual location tracking application developed and trialed at Carnegie Mellon University (Pittsburgh, USA) was used as an example. The application, called PeopleFinder, allows users to share their current location with friends and colleagues, based on a set of location disclosure rules, e.g., ‘‘Jennifer can see my location with high accuracy only on weekdays.’’ The authors were particularly concerned with the usability S. Spiekermann (&) Institute of Information Systems, Humboldt University Berlin, Berlin, Germany e-mail: sspiek@wiwi.hu-berlin.de

Comparative Proteomics Approach to Screening of Potential Diagnostic and Therapeutic Targets for Oral Squamous Cell Carcinoma

This work demonstrates that a comprehensive strategy of proteomics identification combined with further validation and detailed functional analysis should be adopted in the field of cancer biomarker discovery. A comparative proteomics approach was utilized to identify differentially expressed proteins in 10 oral squamous carcinoma samples paired with their corresponding normal tissues. A total of 52 significantly and consistently altered proteins were identified with eight of these being reported for the first time in oral squamous carcinoma. Of the eight newly implicated proteins, RACK1 was chosen for detailed analysis. RACK1 was demonstrated to be up-regulated in cancer at both the mRNA and protein levels. Immunohistochemical examination showed that the enhanced expression of RACK1 was correlated with the severity of the epithelial dysplasia as well as clinical stage, lymph node involvement, and recurrence, which are known indicators of a relatively poor prognosis in oral squamous carcinoma patients. RNA interference specifically targeted to silence RACK1 could initiate apoptosis of oral squamous carcinoma cells. Taken together, the results indicate that RACK1 is up-regulated in oral squamous carcinoma, not only being closely related to cell proliferation and apoptosis but also linked to clinical invasiveness and metastasis in carcinogenesis. The observations suggest that RACK1 may be a potential biomarker for early diagnosis, prognosis, and monitoring in the therapy of oral squamous carcinoma. Further this comprehensive strategy could be used for identifying other differentially expressed proteins that have potential to be candidate biomarkers of oral squamous carcinoma.

Breathing in Granular Chains and Conversion of Wave Energy to Useful Forms

Estimates suggest that a meter of a wave front carries ̃ 100 kW of power. A significant amount of renewable energy is hence dissipated across the time scale of minutes as surface gravity waves dissipate on any beach. Given that the majority of the world population lives near the ocean, it makes sense to explore the conversion of wave energy into electrical energy. Here we consider a new concept - that of a potentially scalable system that converts wave energy into energy in mechanical pulses and then to voltage drops. The envisioned system would exploit the well studied physics of nonlinear repulsion between elastic grains encased in cylinders to continuously convert the incident wave energy into dispersionless energy bundles, which can be subsequently converted to stress pulses in a piezo-electric slab. The key points involved in the concept will be presented.

Metastin Inhibits Migration and Invasion of Renal Cell Carcinoma with Overexpression of Metastin Receptor

BackgroundMetastin, the final peptide of the KiSS-1 gene, has been proposed to suppress cell motility. ObjectiveThis study investigated whether renal cell carcinoma (RCC) tissue expresses metastin or its receptor, and clarified whether metastin can suppress migration and/or invasion and/or proliferation of RCC cells in vitro. Design, Setting, and ParticipantsTwenty-five RCC samples were submitted. Fresh RCC tissues were prepared for real-time RT-PCR, and formalin-fixed and paraffin-embedded tissues blocks were examined by immunohistochemistry. RCC cell lines Caki-1 and ACHN were supplied for cell migration, invasion, and proliferation assays. MeasurementsReal-time RT-PCR was performed by using Taq Man gene expression system. ENVISION system was used in immunohistochemistry. Wound-healing assay and matrigel assays were used to identify migration and invasion abilities of RCC cell lines. Cell Counting Kit-8 was applied to measure the cell proliferation. Cell morphology was examined under a META system. Statistical analysis was performed with SPSS®15.0J. Results and LimitationsIn twenty-five RCC samples, the mRNA level of metastin receptor was identified to be significantly higher than non-neoplastic renal cortex by real-time RT-PCR (p=0.011). Immunohistochemical study also detected metastin receptor protein in all RCC tumors. In vitro, this study showed that metastin inhibited migration and invasion of Caki-1 and ACHN cells. In contrast, it had no effects on cell proliferation. Metastin (10μmol/l) induced excessive formation of focal adhesions and stress fibers in Caki-1 and ACHN cells; this phenomenon was inhibited by pretreating pharmacological Rho-kinase inhibitor (Y-27632) to those cells. ConclusionThis is the first report regarding overexpression of the metastin receptor hOT7T175 in human RCC. We demonstrate that metastin can inhibit migration and invasion of the RCC cell line, which is regulated by a Rho-kinase inhibitor. Metastin and its receptor are therefore probable targets for suppressing RCC.