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Introduction:To explore the role of computed tomography (CT) texture analysis in predicting T-stage of gastric cancers (GC).Methods:Preoperative enhanced CT images of 110 patients (men: 84, women: 26) with GC were reviewed retrospectively. Regions of interest were manually drawn along the margin of the lesion on the section where it appeared largest on the portal venous CT images, which yielded texture parameters (1, 10, 50, 90, and 99% percentiles; minimum, mean, and maximum norm; variance; skewness, and kurtosis). Correlations between texture parameters and pathological stage were analysed with Spearman’s correlation test. The distributions of all variables were checked with the aid of the Kolmogorov-Smirnov test. The Independent-Samples t-test and the Mann-Whitney U test were used (as appropriate) to compare quantitative data. The chi-squared test was employed to compare qualitative data. The diagnostic performance of CT texture parameters in differentiating different stages was evaluated using receiver operating characteristic analysis.Results:The T4 variance was significantly greater than that of the T1-to-T3 group (p˂0.05). The T4 skewness was significantly lower than that of the T1-to-T3 group (p˂0.05) but the T4 kurtosis significantly higher (p˂0.05).Conclusion:The histogram parameters of CT-TA, especially skewness and kurtosis derived from portal, venous phase CT images, may serve as biomarkers stratifying the risk of serosal invasion (stage-T4) by locally advanced GC. Thus, histogram analysis can be used preoperatively to evaluate serosal invasion.

International audience; Curcuminoids, which include natural acyclic diarylheptanoids and the synthetic analogs of curcumin, have considerable potential for fighting against all the characteristics of invasive cancers. The epithelial-to-mesenchymal transition (EMT) is a fundamental process for embryonic morphogenesis, however, the last decade has confirmed it orchestrates many features of cancer invasiveness, such as tumor cell stemness, metabolic rewiring, and drug resistance. A wealth of studies has revealed EMT in cancer is in fact driven by an increasing number of parameters, and thus understanding its complexity has now become a cornerstone for defining future therapeutic strategies dealing with cancer progression and metastasis. A specificity of curcuminoids is their ability to target multiple molecular targets, modulate several signaling pathways, modify tumor microenvironments and enhance the host's immune response. Although the effects of curcumin on these various parameters have been the subject of many reviews, the role of curcuminoids against EMT in the context of cancer have never been reviewed so far. This review first provides an updated overview of all EMT drivers, including signaling pathways, transcription factors, non-coding RNAs (ncRNAs) and tumor microenvironment components, with a special focus on the most recent findings. Secondly, for each of these drivers the effects of curcumin/curcuminoids on specific molecular targets are analyzed. Finally, we address some common findings observed between data reported in the literature and the results of investigations we conducted on experimental malignant mesothelioma, a model of invasive cancer representing a useful tool for studies on EMT and cancer.

Gastric cancer remains a disease with a dismal prognosis. Extensive efforts to find targetable disease drivers in gastric cancer were implemented to improve patient outcomes. Beyond anti-HER2 therapy, MET pathway seems to be culprit of cancer invasiveness with MET-overexpressing tumors having poorer prognosis. Tyrosine kinase inhibitors targeting the HGF/MET pathway were studied in MET-positive gastric cancer, but no substantial benefit was proven. Some patients responded in early phase trials but later developed resistance. Others failed to show any benefit at all. Etiologies of resistance may entail inappropriate patient selection with a lack of MET detection standardization, tumor alternative pathways, variable MET amplification, and genetic variation. Optimizing MET detection techniques and better understanding the MET pathway, as well as tumor bypass mechanisms, are an absolute need to devise means to overcome resistance using targeted therapy alone, or in combination with other synergistic agents to improve outcomes of patients with MET-positive GC.

The severity of the bladder carcinoma (BC) is directly linked to cell invasion and metastasis. Indoleamine 2,3-dioxygenase-1 (IDO-1) is an INF-γ-induced immunomodulating enzyme that has been linked to the cancer cell invasiveness. Because IDO1 is variable among the tumors, we analyzed its expression in the BC invasion using BC mice models and cell culture. MB49 cells were orthotopically or ectopically inoculated in C57Bl6 mice to evaluate IDO1 by immunohistochemistry. For in vitro experiments, expression of IDO1 and INF-γ was evaluated in grade-1 (RT4) and in grade-3 (T24) BC cell lines. Invading and non-invading T24 cells were separated using the Matrigel/Transwell system, of which total RNA was extracted immediately or after 2 weeks of subculture. Finally, IDO1 was silenced in T24 cells to verify its role on cell invasiveness. In both animal models, IDO1 was differentially expressed between non-invading and invading cells. In cell culture, T24 cells expressed more IDO1 than RT4 cells, independently of the INF-γ expression. IDO1 was differentially expressed between non-invading and invading T24 cells, a difference that was lost by long-time subculture. IDO1 silencing resulted in diminished cell invasiveness. In conclusion, IDO1 expression is changed during bladder carcinoma invasion, playing an important role in this process.

Metastasis is the primary cause of death from cancer. Current lack of medicines for metastasis makes it a topical issue and encourages the global search for molecules that may potentially inhibit invasiveness of cancer cells, and thus prevent metastatic spread. Drug repurposing is a strategy of identifying new uses for approved drugs that are outside the scope of the original medical indication. The aim of this thesis is to summarize knowledge on the repurposing potential of selected groups of drugs with emphasis on molecular mechanisms of their action. In particular, the work is focused on selected antipsychotics, antidepressants, beta- blockers and statins, whose abilities to inhibit cancer invasiveness have been indicated.

uncorrected proof version, The generation of invasive fluctuating protrusions is a distinctive feature of tumor dissemination. During the invasion, individual cancer cells modulate the morphodynamics of protrusions to optimize their migration efficiency. However, it remains unclear how protrusion fluctuations govern the invasion of more complex multi-cellular structures, such as tumors, and their correlation with the tumor metastatic potential. Herein, a reductionist approach based on 3D tumor cell micro-spheroids with different invasion capabilities is used as a model to decipher the role of tumor-associated fluctuating protrusions in cancer progression. To quantify fluctuations, a set of key biophysical parameters that precisely correlate with the invasive potential of tumors is defined. It is shown that different pharmacological drugs and cytokines are capable of modulating protrusion activity, significantly altering protrusion fluctuations, and tumor invasiveness. This correlation is used to define a novel quantitative invasion index encoding the key biophysical parameters of fluctuations and the relative levels of cell-cell/matrix interactions, which is capable of assessing the tumor's metastatic capability solely based on its magnitude. Overall, this study provides new insights into how protrusion fluctuations regulate tumor cell invasion, suggesting that they may be employed as a novel early indicator, or biophysical signature, of the metastatic potential of tumors., Portuguese Foundation for Science and Technology (FCT) under the program CEEC Individual 2017 (CEECIND/00352/2017). D.C., V.B., A.C.L., C.M.A., and S.C.K. also thank the support from the FCT under the scope of the projects 2MATCH (PTDC/BTM-ORG/28070/2017) and BREAST-IT (PTDC/ BTM-ORG/28168/2017) funded by the Programa Operacional Regional do Norte supported by European Regional Development Funds (ERDF). The authors also thank the financial support from the European Union Framework Program for Research and Innovation Horizon 2020 on FoReCaST project under (Grant Number: 668983), and the Institution of Engineering and Technology (IET) for the funding provided under the ENG ThE CANCER project (Grant Number: IET Harvey Engineering Research Award 2018). J.M.O. is grateful for the FCT distinctions (IF/00423/2012 and IF/01285/2015)

A major factor for developing new tumor models is to recreate a proper three-dimensional environment for 3D tumors culture. In this 3D microenvironment, extracellular matrices play important roles in regulation of hallmark features of cancer through biochemical and mechanical signals. The fabrication of a mechanical and biophysical controllable hydrogel, while sharing similarities with Matrigel in cancer invasiveness evaluation, is an urgent but unmet need. In this study, we developed a hybrid hydrogel system composed of GelMA and hydrolyzed collagen to model tumor micro-environment and tested with several cancer cells with different origin and characteristics. This hydrogel possesses a well-ordered homogenous microstructure, excellent permeability and an adjustable mechanical stiffness. This hydrogel demonstrated similar properties as Matrigel in tumor spheroids culture and 3D tumor invasiveness studies. It was further applied in a Tumor-on-a-Chip system with 3D-bioprinting. Our research demonstrated this hydrogel's effectiveness in tumor 3D culture, and its potential to replace Matrigel in cancer invasiveness evaluation.

Background: Brain metastasis is a major cause of cancer patient morbidity and mortality. The late stage of diagnosis and the difficulty in defining the tumour boundary both contribute to the lethality of brain metastases. Cell adhesion molecules (CAMs), such as very late antigen-4 (VLA-4) and its ligand vascular cell adhesion molecule-1 (VCAM-1), have been implicated in the metastatic process. By targeting the VLA-4/VCAM-1 interaction, I applied different methods to improve the diagnosis of brain metastases and to prevent their formation. Aims: (i) To determine if VCAM-1 targeted MRI is able to detect brain micrometastases from different tumour types in mouse models; (ii) to determine if VCAM-1 targeted MRI is able to detect the brain metastasis invasive margin in rat models; and (iii) to determine if tumour cell VLA-4 knockdown inhibits metastatic spread of cancer to the brain in mouse models. Findings: VCAM-MPIO MRI could detect brain metastases of different tumour types before gadolinium enhanced T1-weighted MRI. Furthermore, the imaging contrast improved delineation of the invasive margin of brain macrometastases on T2*-weighted MRI, when used in conjunction with conventional techniques. Whilst knockdown of VLA-4 does not entirely prevent brain metastasis formation, the trend to benefit suggests testing in combination with knockdown of other CAMs may yield a significant reduction in metastatic potential. Discussion: I present a multi-faceted approach, exploiting the roles of VLA-4 and VCAM-1 in brain metastatic formation and progression, for improved diagnosis of brain micro/macrometastases and possible inhibition of metastatic seeding in the brain. Future studies will focus on improving the preventative capacity of anti-CAM therapy and the clinical translation of VCAM-MPIO MRI to human applications.

The overproduction and deposition of hyaluronic acid (HA) of different sizes in the tumor microenvironment is associated with cancer metastasis. Here, the development of layerâ byâ layer (LbL) constructs containing HA of different molecular weights (i.e., 5.6, 618, and 1450Â kDa) that mimic the HAâ rich cancer extracellular matrix is described to study the effect of the HA's size on the behavior of gastric cancer cells (AGS). The results demonstrate that LbL constructs with short HA, i.e., 5.6Â kDa, activate the cytoskeleton rearrangement leading to the â hummingbirdâ morphology, promote high cellular motility, and activate signaling pathways with increased expression of pâ ERK1/2 and pâ AKT. In addition, it is demonstrated that this malignant transformation involves an active participation of the HA coreceptor RHAMM in AGS cells., The authors acknowledge the financial support from the European Commission’s H2020 Programme, under grant agreements H2020-WIDESPREAD-2014-668983-FORECAST and H2020-MSCA-RISE-2019-872648-MEPHOS. S.A. acknowledge the Portuguese Foundation for Science and Technology (FCT) for the PhD grant (SFRH/BD/112075/2015). The authors also thank Ramon Novoa-Caballal for performing the GPC experiments and data analysis.

e18063 Background: Epithelial-to- mesenchymal transition (EMT) plays a key role not only in cancer invasiveness and progression but also in treatment resistance. Experimental models suggest that epithelial and mesenchymal phenotypes may represent steady states supported by the regulatory logic of intracellular signaling networks. Using known regulatory interactions between intracellular signaling molecules as a framework, we explored EMT in silico with the aim of identifying potentially novel means of inhibiting metastasis in ovarian cancer. Methods: A mathematical model connecting 27 genes involved in EMT through 153 regulatory interactions was assembled from the Mogrify, Reactome, String and Pathway Studio (PS) databases, the last of which used the MedScan natural language processing (NLP) engine to process over 8,000 full-text publications. Logic model parameters dictating the regulatory response dynamics of the EMT circuit were constrained to values that accurately predicted the proteomic profiles characterized as either epithelial or mesenchymal in 30 cell lines in the publicly available MaxQB database. Results: A total of 94,672 candidate models predicted the protein expression in stable epithelial and mesenchymal cells equally well, clustering into 24 classes of slightly different EMT transition kinetics. Discrete event simulations based on these models identified 7 unique combinations, involving the concurrent modulation of at least 3 gene products capable of effecting this transition successfully even in the presence of noise. Every combination required inactivation of ZEB2, along with 2 other manipulations. Inactivation of TP53, HOXA5, or IRF7 was required in 3 combinations, while inactivation of ICAM1 was required in 2 cases. Only BCL2 was required to be activated in any of these scenarios. Interestingly, BCL2 inhibition is now applied clinically in some hematologic cancers and proving effective in patient-derived cell lines for high-grade serous ovarian cancer when combined with MEK inhibition. Conclusions: This work suggests that EMT might at least in part be driven by normal regulatory interactions between specific proteins in a network of pathways, without the requirement for additional mutations or alterations in the underlying circuitry. Maintenance of ZEB2 or TP53 activation as well as BCL2 inhibition may thus represent promising avenues for future research in arresting EMT and associated chemo-resistance.

Invadopodia are micron-sized invasive structures that mediate extracellular matrix (ECM) degradation through a combination of membrane-bound and soluble matrix metalloproteinases (MMPs). However, how such localized degradation is converted into pores big enough for cancer cells to invade, and the relative contributions of membrane-bound versus soluble MMPs to this process remain unclear. In this article, we address these questions by combining experiments and simulations. We show that in MDA-MB-231 cells, an increase in ECM density enhances invadopodia-mediated ECM degradation and decreases inter-invadopodia spacing. ECM degradation is mostly mediated by soluble MMPs, which are activated by membrane-bound MT1-MMP. We present a computational model of invadopodia-mediated ECM degradation, which recapitulates the above observations and identifies MMP secretion rate as an important regulator of invadopodia stability. Simulations with multiple invadopodia suggest that inter-invadopodia spacing and MMP secretion rate collectively dictate the size of the degraded zones. Taken together, our results suggest that for creating pores conducive for cancer invasion, cells must tune inter-invadopodia spacing and MMP secretion rate in an ECM density-dependent manner, thereby striking a balance between invadopodia penetration and ECM degradation.

Ovarian cancer is the 5th leading cause of cancer death in women in the United States and is the most fatal gynecological malignancy. High grade serous ovarian cancer (HGSC) is the most common and deadly type of ovarian cancer largely due to the rapid metastasis throughout the peritoneum (abdominal cavity wall and organ lining). Metastatic spread of ovarian cancer usually occurs before diagnosis and can lead to bowel obstruction, organ failure, ascites, cachexia, infection and sepsis, and pulmonary embolism all causing death. Current methods to detect early stage ovarian cancer do not increase overall survival. A better understanding of the metastatic ability of ovarian cancers and the mechanism of cancer cell dissemination are critical to the development of new treatments for this devastating disease. In particular, investigation of pathways that affect early metastasis may indicate treatments that will lower disease burden and may suggest biomarkers of recurrent and/or chemotherapy resistant disease. Notch3 expression correlates with worse prognosis, chemotherapy resistance, and increased tumorigenic cell behaviors in HGSC. Here, we demonstrate that Notch3 acts to promote early stages of metastasis in a model of HGSC using the murine ID8 IP2 ovarian surface epithelial cell line. ID8 IP2 cells have little to no endogenous Notch3 expression and model metastatic disease when introduced intraperitoneally. We investigated the role of Notch3 by ectopically expressing the intracellular domain of murine Notch3 to induce constitutive Notch3 signaling in ID8 IP2 cells and verified Notch signal activation by target gene assessment. Induction of Notch3 signaling in ID8 IP2 reduced survival and accelerated disease burden, as measured by ascites accumulation, after intraperitoneal introduction of cells into nude mice. We interrogated downstream targets in Notch3 activated cells by RNA-Seq and found that Notch3 induced a significant enrichment of adhesion and extracellular matrix pathways. Notch3 active cells showed increased ITGA1 expression and increased adhesion on collagens I and IV in vitro, suggesting that increased adhesion to collagen-rich peritoneal surfaces drives the observed increase in tumor burden. Notch3 active cells showed reduced migration on surfaces coated with multiple types of extracellular matrix and no detectable increase in invasion through extracellular matrix, indicating that Notch3 effects may be specific to the initial adhesion of tumor cells and not the later stages of metastasis. These results demonstrate that Notch3 upregulates the expression of specific adhesion genes in ovarian cancer cells and this promotes increased attachment to the collagen-rich extracellular matrix. The implications of this study are that oncogenic Notch signal activation, as documented in human disease, may promote dissemination and metastasis of primary and/or recurrent HGSC by increasing attachment to the peritoneal lining.

A 64-year old lady presented with worsening abdominal pain, vomiting and constipation. She had diffuse abdominal tenderness with peritonism, requiring an emergency laparotomy as a result of a perforated sigmoid tumour. An incidental hard lump was identified on the anterior surface of the pancreas and was removed by enucleation. It was later diagnosed as metastatic leiomyosarcoma based on histology and from her history of uterine malignancy. No recurrence is reported up to this day. Uterine leiomyosarcomas are aggressive malignant tumours with a high predisposition to metastasis, commonly to the lungs, liver, brain and bone. Metastasis to the pancreas is a rare occurrence and considered highly unusual, which can typically present with non-specific symptoms and signs. Imaging can pick up a pancreatic lesion and a radiologically-guided FNA as a pre-operative attempt is acceptable in order to differentiate the lesion before undertaking any major surgery. However, in view of only a few case reports found in the literature, surgical management of pancreatic metastases is not clearly defined with a questionable long-term prognosis. Most cases are managed by elective radical excision with good result. Review of the literature shows other more radical surgical approaches were used. This is the first report of metastatic uterine leiomyosarcoma to the pancreas being managed by enucleation, with a successful follow-up and no recurrence., peer-reviewed

Thesis (PhD(Biomaterials Engineering and Nanomedicine))--University of South Australia, 2016. Includes bibliographical references (pages 211-212) The presence of disseminated tumour cells (DTCs) in blood (known as circulating tumour cells; CTCs) and in lymph nodes is an indicator of cancer spread throughout the body and is consequently associated with poorer patient prognosis. DTCs are typically rare and therefore, sensitive detection strategies are required. For instance, as little as 1 CTC can be present in a million white blood cells, and potentially less than 1,000 DTC can be present in a lymph node. The rarity of these cells means that either high throughput detection technologies and/or enrichment strategies are vital for reliable identification and accurate characterisation.

To invade and metastasize to distant loci, breast cancer cells must breach the layer of basement membrane surrounding the tumor and then invade through the dense collagen I-rich extracellular environment of breast tissue. Previous studies have shown that breast cancer cell aggregate morphology in basement membrane extract correlated with cell invasive capacity in some contexts. Moreover, cell lines from the same aggregate morphological class exhibited similarities in gene expression patterns. To further assess the capacity of cell and aggregate morphology to predict invasive capacity in physiologically relevant environments, six cell lines with varied cell aggregate morphologies were assessed in a variety of assays including a 3D multicellular invasion assay that recapitulates cell-cell and cell-environment contacts as they exist in vivo in the context of the primary breast tumor. Migratory and invasive capacities as measured through a 2D gap assay and a 3D spheroid invasion assay reveal that breast cancer cell aggregate morphology alone is insufficient to predict migratory speed in 2D or invasive capacity in 3D. Correlations between the 3D spheroid invasion assay and gene expression profiles suggest this assay as an inexpensive functional method to predict breast cancer invasive capacity.

To better understand the progression of heterogeneous breast cancers, four models of progression pathways have been evaluated. The models describe the progression through the grades of ductal carcinoma in situ (DCIS) 1, 2, and 3, and through the grades of invasive ductal carcinoma (IDC) 1, 2, and 3. The first three pathways, termed linear, nonlinear, and branched, describe DCIS as aprogenitor of IDC, and grades of DCIS progressing into grades of IDC. The fourth pathway, termed parallel, describes DCIS and IDC as diverging from a common progenitor and progressing through grades in parallel. The best transition rates for the linear, nonlinear, and branched pathways were sought using a random search in combination with a directed search based on the Nelder���Mead simplex method. Parameter values for the parallel pathway were determined with heuristic graphs. Results of computer simulation were compared with clinically observed frequencies of grades of DCIS and grades of IDC that were reported to occur together in heterogeneous tumors. Each of the four pathways could simulate frequencies that resembled, to varying degrees, the clinical observations. The parallel pathway produced the best correspondence with clinical observations. These results quantify the traditional descriptions in which grades of DCIS are the progenitors of grades of IDC. The results also raise the alternative possibility that, in some tumors with both components, DCIS and IDC may have diverged from a common progenitor.

Einleitung Bei der Entschlusselung molekularer Grundlagen des Tumor-wachstums und der Tumorprogression wurden in den letzten Jahren wesentliche Fortschritte erzielt und wichtige Mechanis-men malignen Wachtums identifiziert. Die Gefasneubildung wurde dabei als ein essentieller Prozess der Tumordisseminie-rung erkannt, ohne den wachsende Tumoren von der Nahrstoff-versorgung abgeschnitten waren. Angiogenese wird uber eine Vielzahl pro- und antiangiogener Faktoren reguliert, die in einem komplexen Wechselspiel ineinandergreifen. Dem VEGF (vascular endothelial growth factor) kommt dabei zusammen mit seinen Rezeptoren (VEGFR) eine Schlusselrolle als poten-ter Mediator der Gefasneubildung zu. VEGF wird von zahl-reichen Tumoren uberexprimiert, ein Charakteristikum, das mit erhohter Tumorinvasivitat und Metastasierung sowie einer schlechten Prognose einhergeht. Eine Hemmung der uber VEGF vermittelten Signaltransduktion durch Neutralisierung des Wachstumsfaktors oder durch Rezeptor-Blockade wird daher seit einigen Jahren als interessantes Ziel zur Entwicklung neuer Therapiestrategien betrachtet. Mittlerweile konnte die Bedeutung dieses Ansatzes klinisch bestatigt werden.

İlaçlara kattığı yüksek çözünürlük, stabilite ve zaman salınımı gibi özellikleri olması itibariyle lipozomlar etkili nano taşıyıcılar olarak yaygınca kullanılmaktadır. Toksik etki göstermemeleri, biyobozunurluğu ve hücre zarına benzer yapıda olmaları da lipozomların tercih edilmesindeki diğer sebeplerdir. Buna rağmen lipozomların hücre içine alınımının kısıtlı olması, lipozomların ilaç taşıyıcısı olarak kullanımında önemli sorunlar teşkil etmektedir. Bu durumun üstesinden gelme amacıyla lipozomları reseptör ve yük bağıntılı işlev gören hücre delici peptitlerle modifiye edip lipozomların hücre içine girişini desteklemek kayda değer bir stratejidir.Bu çalışmada, liposomları tek adımda modifiye eden bir sistem geliştirilmiş olup, entegre olan amfifilik hücre delici peptitlerin lipozomların etkinliği üzerindeki rolü test edilmiştir. Arjinin miktarı itibariyle zengin hücre delici bölümü ve hidrokarbon zincirine sahip hücre delici peptitler DOPG ve kolesterolden oluşan eksi yüklü lipozomlara entegre edilmiştir. Çalışmada modifiye edilmiş lipozomların (a) boyut, yüzey potansiyeli ve zar polaritesi gibi fiziksel özellikleri, (b) hidrofilik Rhodamin B ve hidrofobik Nile Red gibi ilaç modellerinin enkapsülasyon ve iletim etkinliği, (c) Paclitaxel ve Doxurobicin-HCl gibi anti kanser ilaçlarla yüklenmesi sonrası terapütik etkinliği araştırılmıştır. Sonuçlar ışığında amfifilik hücre delici peptitlerle kovalent bağ olmaksızın modifiye edilmiş lipozomların ilaç iletimi yönünden daha iyi olup, anti kanser ilaçlarla yüklendiğinde terapütik etkinliğini arttığını göstermiştir. Liposomes have been extensively utilized as effective nanocarriers due to their enhanced solubility, higher stability and greater ability to facilitate the slow release of encapsulated drugs compared to free drug administrations. Liposomes are also preferred as drug vectors due to their non-toxic nature, biodegradability and structural resemblance to the cell membrane. However, their low internalization efficiencies pose an important challenge for their use in drug delivery applications. Internalization issues inherent in many liposomal systems can be circumvented by the use of cell penetrating peptides, which non-covalently attach on the liposome surface and greatly enhance liposomal uptake in a receptor- and charge-dependent manner.In this study, we examined the liposomal dynamics effected through the integration of an amphiphilic cell penetrating peptide into a simple liposome system. Peptide amphiphiles with a cell penetrating arginine-rich domain were incorporated into liposomal membranes formed by negatively charged dioleoylphosphoglycerol (DOPG) phospholipids in the presence of cholesterol. Throughout the present study, we sought to analyze the effect of peptide incorporation on (a) the physical characteristics, such as size, surface potential and membrane polarity, of the liposomal membrane, (b) the alterations in the encapsulation and delivery mechanisms of hydrophilic (Rhodamine B) and hydrophobic (Nile Red) drug models and (c) the enhancement of therapeutic capability in liposomes loaded with the drugs Doxorubicin-HCl and Paclitaxel. Our results revealed that the modification of liposomes by cell penetrating peptide amphiphiles results in the improvement of cargo delivery and the enhancement of the therapeutic effects of the anti-cancer drugs Doxurubicin and Paclitaxel. 95

Background The biological mechanisms underlying cancer cell motility and invasiveness remain unclear, although it has been hypothesized that they involve some type of epithelial-mesenchymal transition (EMT). Methods We used xenograft models of human cancer cells in immunocompromised mice, profiling the harvested tumors separately with species-specific probes and computationally analyzing the results. Results Here we show that human cancer cells express in vivo a precise multi-cancer invasion-associated gene expression signature that prominently includes many EMT markers, among them the transcription factor Slug, fibronectin, and α-SMA. We found that human, but not mouse, cells express the signature and Slug is the only upregulated EMT-inducing transcription factor. The signature is also present in samples from many publicly available cancer gene expression datasets, suggesting that it is produced by the cancer cells themselves in multiple cancer types, including nonepithelial cancers such as neuroblastoma. Furthermore, we found that the presence of the signature in human xenografted cells was associated with a downregulation of adipocyte markers in the mouse tissue adjacent to the invasive tumor, suggesting that the signature is triggered by contextual microenvironmental interactions when the cancer cells encounter adipocytes, as previously reported. Conclusions The known, precise and consistent gene composition of this cancer mesenchymal transition signature, particularly when combined with simultaneous analysis of the adjacent microenvironment, provides unique opportunities for shedding light on the underlying mechanisms of cancer invasiveness as well as identifying potential diagnostic markers and targets for metastasis-inhibiting therapeutics.

Nano-mechanical properties of prostate cancer cells with different metastatic potentials were investigated, in vitro, to shed light on the issue whether those properties could be utilized as indicators of their aggressiveness. Experimentations involved atomic force microscopy-based indenting and force mapping experiments acquired at LNCaP (lowly metastatic) and CL-1 (highly metastatic) prostate cancer cells that were cultured on plain glass and nano-scaffolds. Various models were applied to extract the mechanical information pertaining to the elasticity, adhesion and topography of the cells. Deviating from the general perspective for metastasis, the elasticity measurements reveal that the elastic modulus of highly metastatic cancer cells, CL-1, is about twice as high (i.e., stiffer) as the elastic modulus of lowly metastatic cancer cells, LNCaP, both at the center of cells’ body and lamellipodia. The two-dimensional maps of adhesion property, in conjunction with the results from the experiments conducted on the nano-scaffolds, reveal that the stiffer cells, CL-1, are more adherent than softer cells, LNCaP. These findings are consistent to the reports observed at in vitro nano-mechanical studies that cultured adherent cells are less deformable than the non-adherent ones. We postulate that the enhanced adhesion generates larger cortical tension that leads to higher elastic modulus observed in CL-1.

Imaging is key in the accurate monitoring of response to cancer therapies targeting tumour vascularity to inhibit its growth and dissemination. Dynamic contrast enhanced ultrasound (DCE ultrasound) is a quantitative method with the advantage of being non-invasive, widely available, portable, cost effective, highly sensitive and reproducible using agents that are truly intravascular. Under the auspices of the initiative of the Experimental Cancer Medicine Centre Imaging Network, bringing together experts from the UK, Europe and North America for a 2-day workshop in May 2010, this consensus paper aims to provide guidance on the use of DCE ultrasound in the measurement of tumour vascular support in clinical trials. Key Points • DCE ultrasound can quantify and extract specific blood flow parameters, such as flow velocity, relative vascular volume and relative blood flow rate. • DCE ultrasound can be performed repeatedly and is therefore ideally suited for pharmacokinetic and pharmacodynamic studies evaluating vascular-targeted drugs. • DCE ultrasound provides a reproducible method of assessing the vascular effects of therapy in pre-clinical and early clinical trials, which is easily translatable into routine clinical practice. ABSTRACT FROM AUTHOR] Copyright of European Radiology is the property of Springer Science & Business Media B.V. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) 22 1442

Antisens oligodeoksinükleotitler (ODN) kanser dâhil birçok hastalığın tedavisinde kullanılabilen önemli ilaçlardır. Ancak kimyasal dayanıklılık, kontrollü salım ve hücre içerisine alım oligonükleotitlerin verimini etkileyen önemli faktörlerdir. Nanofibrillerden oluşan üç boyutlu ağsı yapıya sahip jeller ilaçların kontrollü salımı ve taşımı amacıyla ilaç dayanıklılığını arttırmak için kullanılmıştır, fakat oligonükleotitler için bir çalışma yoktur. Bu çalışmada, programlı toplanabilir peptit nanofiber sistemi, pozitif yüklü amfifilik peptit (AP) molekülleri ile Bcl-2 antisens oligodeoksinükleotidi, G3139, karıştırılarak elektrostatik etkileşimlerin etkisiyle oluşturuldu. Programlı toplanabilir AP ve ODN jeli dairesel dikroism, reoloji, atomik kuvvet mikroskobu (AFM) ve taramalı elektron mikroskobu (SEM) kullanılarak karakterize edildi. AFM ve SEM görüntüleri jelin nanofiber ağsı yapısını göstermektedir. Jelden ODN salımı katyonik AP'ler ve anyonik ODN'ler arasındaki elektrostatik etkileşimlere bağlıdır. Çalışmamızda AP ve ODN konsantrasyonları değiştirilerek ODN salımı kontrol edildi. Floresan işaretli ODN kullanılarak AP-ODN kompleksinin hücre içerisine alınımı incelendi. AP-ODN kompleksi uygulanan MCF-7 insan meme kanseri hücrelerinin daha fazla floresan sinyali verdiği gözlendi. Ayrıca AP-ODN kompleksi ile inkübe edilen MCF-7 hücrelerinin Bcl-2 mRNA ifadesinin, çıplak halde ODN ve 2-bazı eşleşmeyen (mismatch-MM) ODN, 2-bazı eşleşmeyen (mismatch-MM) ODN ve AP kompleksi ile inkübe edilen hücrelerdeki mRNA ifadesinden daha fazla azaldığı kantitatif RT-PZR yöntemi ile gösterildi. Bu sonuçlar AP moleküllerinin ODN salımının kontrolü ve hücre içerisine verimli ODN alımı için kullanılabileceğini ve gen terapisinde yeni bir yaklaşım olabileceğini önermektedir. Devam eden çalışmalarda, ODN`nin hücre içerisine alımının RGDS biyoaktif dizileriyle arttırıldığı gösterildi. Antisens oligodeoksinükleotitler (ODN) kanser dâhil birçok hastalığın tedavisinde kullanılabilen önemli ilaçlardır. Ancak kimyasal dayanıklılık, kontrollü salım ve hücre içerisine alım oligonükleotitlerin verimini etkileyen önemli faktörlerdir. Nanofibrillerden oluşan üç boyutlu ağsı yapıya sahip jeller ilaçların kontrollü salımı ve taşımı amacıyla ilaç dayanıklılığını arttırmak için kullanılmıştır, fakat oligonükleotitler için bir çalışma yoktur. Bu çalışmada, programlı toplanabilir peptit nanofiber sistemi, pozitif yüklü amfifilik peptit (AP) molekülleri ile Bcl-2 antisens oligodeoksinükleotidi, G3139, karıştırılarak elektrostatik etkileşimlerin etkisiyle oluşturuldu. Programlı toplanabilir AP ve ODN jeli dairesel dikroism, reoloji, atomik kuvvet mikroskobu (AFM) ve taramalı elektron mikroskobu (SEM) kullanılarak karakterize edildi. AFM ve SEM görüntüleri jelin nanofiber ağsı yapısını göstermektedir. Jelden ODN salımı katyonik AP?ler ve anyonik ODN?ler arasındaki elektrostatik etkileşimlere bağlıdır. Çalışmamızda AP ve ODN konsantrasyonları değiştirilerek ODN salımı kontrol edildi. Floresan işaretli ODN kullanılarak AP-ODN kompleksinin hücre içerisine alınımı incelendi. AP-ODN kompleksi uygulanan MCF-7 insan meme kanseri hücrelerinin daha fazla floresan sinyali verdiği gözlendi. Ayrıca AP-ODN kompleksi ile inkübe edilen MCF-7 hücrelerinin Bcl-2 mRNA ifadesinin, çıplak halde ODN ve 2-bazı eşleşmeyen (mismatch-MM) ODN, 2-bazı eşleşmeyen (mismatch-MM) ODN ve AP kompleksi ile inkübe edilen hücrelerdeki mRNA ifadesinden daha fazla azaldığı kantitatif RT-PZR yöntemi ile gösterildi. Bu sonuçlar AP moleküllerinin ODN salımının kontrolü ve hücre içerisine verimli ODN alımı için kullanılabileceğini ve gen terapisinde yeni bir yaklaşım olabileceğini önermektedir. Devam eden çalışmalarda, ODN`nin hücre içerisine alımının RGDS biyoaktif dizileriyle arttırıldığı gösterildi. 81

Background The biological mechanisms underlying cancer cell motility and invasiveness remain unclear, although it has been hypothesized that they involve some type of epithelial-mesenchymal transition (EMT). Methods We used xenograft models of human cancer cells in immunocompromised mice, profiling the harvested tumors separately with species-specific probes and computationally analyzing the results. Results Here we show that human cancer cells express in vivo a precise multi-cancer invasion-associated gene expression signature that prominently includes many EMT markers, among them the transcription factor Slug, fibronectin, and α-SMA. We found that human, but not mouse, cells express the signature and Slug is the only upregulated EMT-inducing transcription factor. The signature is also present in samples from many publicly available cancer gene expression datasets, suggesting that it is produced by the cancer cells themselves in multiple cancer types, including nonepithelial cancers such as neuroblastoma. Furthermore, we found that the presence of the signature in human xenografted cells was associated with a downregulation of adipocyte markers in the mouse tissue adjacent to the invasive tumor, suggesting that the signature is triggered by contextual microenvironmental interactions when the cancer cells encounter adipocytes, as previously reported. Conclusions The known, precise and consistent gene composition of this cancer mesenchymal transition signature, particularly when combined with simultaneous analysis of the adjacent microenvironment, provides unique opportunities for shedding light on the underlying mechanisms of cancer invasiveness as well as identifying potential diagnostic markers and targets for metastasis-inhibiting therapeutics.

Bizim bu projedeki amacımız, anti kanser özelliğe sahip olabilecek molekülleri test edip nitelendirmektir.İncelediğimiz ilk grup, yüksük otundan elde edilen kalp glikosidleriydi. Bu moleküller, gerek kalp hastalığı tedavisinde, gerekse de kanser tedavisindeki potansiyel etkileri sebebiyle oldukça araştırılmakta olan moleküllerdir. Bizim araştırmalarımız sonucunda da bu maddelerin karaciğer kanseri hücrelerine sitotoksik etkileri olduğunu göstermiş bulunmaktayız. Ayrıca bu moleküller, hücre bölünme sürecinde G2/M fazında durmaya sebep vererek apoptos indüklenmesine neden oldukları deney sonuçları da gösterilmiştir.İkinci grup moleküllerimiz sentetik kinaz inhibitör türevleridir. Günümüzde kinaz inhibitörü olan imatinib, erlotanib gibi birçok madde kanser tedavisinde kullanılmaktadır. Biz de yaptığımız araştırmalar sonucunda gruptaki 16 molekülden özellikle bir molekülün oldukça yüksek citotoksik etki ve kinas inhibisyon gücünün varlığını göstermiş olduk.Son grup moleküller ise tiyazolidin yapısındaki moleküllerden oluşmaktadır. Bu grupta da özellikle bir molekülün öne çıktığını gösterdik. Alkin içeren bu molekül oldukça yüksek hücre ölümlerine neden oluyor kanser hücre ortamına verildiğinde. Ayrıca bu molekülün de hücre bölünme döngüsünde SubG1/G1 fazında durmaya sebep olduğunu deney sonuçlarımızda gösterdik. The ultimate goal of our project was to investigate candiate small molecules with a potential anticancer activity and characterize their mode of action.Cardiac glycosides are important group of molecules for both their treating properties in heart failure and their potential effects in cancer therapy. We investigated the cardiac glycosides that are extracted from Digitalis Ferruginea which can be frequently found in Turkey. These glycosides are Lanatoside A, Lanatoside C and Glucogitorosid. Our results showed that they constitute high cytotoxicity effect against liver cancer cell lines. In addition they cause G2/M cell cycle arrest and thereby induce apoptosis.For the synthetic molecules, we first tested a set of molecules that are synthesized as derivatives of kinase inhibitors. There are some commercial drugs such as imatinib or erlotinib that are used frequently for cancer treatment. Thus we wanted to investigate if these molecules comprise cytotoxic activities. Our data revealed that especially one of the molecules out of 16 display high cytotoxicity and high kinase inhibitory effect in liver cancer cell lines.The final group of molecules we tested was compoused of thiazolidine ring. In this group of molecules, only one molecule, the one with alkyne terminal precursor, caused cytotoxicity against cancer cell lines. Besides, we have shown that it induces SubG1/G1 cell cycle arrest in cancer cell lines. 92

Cell invasiveness is essential for cancer metastasis. Many proteins, and more recently also non-coding RNAs, particularly microRNAs (miRNAs), have been reported to affect the cell invasiveness of various cancers. There is an apparent gap between the high number of these macromolecules and the low number of signaling pathways experimentally verified to control cancer invasiveness. We have brought together these various proteins and RNAs because we could not find any publication that filled this important gap. We have noted 589 proteins, 28 miRNAs, and 1 long non-coding RNA that are reported to modulate invasiveness in cells of various cancers. Interestingly, 44 proteins enhance invasiveness in cells of some cancers, but suppress it in cells of others. Almost all of the proteins that show experimentally verified activation/inhibition effects on, or binding interactions with, each other are linked together in a single network, in a “hub-and-spoke” architecture. The accumulated data show trends that point to anticipated future results and highlight gaps in what is known about invasiveness signaling. Identification of cancer invasiveness signaling networks is important for combination and personalized targeted therapies of cancers. J. Cell. Biochem. 111: 791–796, 2010. © 2010 Wiley-Liss, Inc.

Tumour metastasis represents the most lethal aspect o f cancer. It involves initial attachment of cells to the extracellular matrix (ECM), degradation of the ECM and subsequent migration and detachment of cells from the ECM. This process depends on alterations in the expression of many key players including cell adhesion molecules (CAMs) and proteolytic enzymes such as the matrix metalloproteinases (MMPs). The MMPs are a large family of proteases collectively capable of degrading the entire ECM. CD44 is a multifunctional transmembrane adhesion molecule, which also plays a key role in cell signalling. The interactions between CAMs and MMPs are crucial therefore in controlling tumour cell invasion and metastasis and this study aimed to elucidate these interactions. We have established a panel of human colon cancer cell lines including a primary human colon adenocarcinoma cell line SW480 and its lymph node metastatic derivative, SW620. The SW480 cells have been stably transfected with the cDNA for MMP-7 (SW480M7) and MMP-9 (SW480M9). When the in vitro invasive activities of the cell lines were compared, the SW620 cells and the MMP transfected cells were more invasive than their parental SW480. The SW480M7 cells were more migratory in comparison to SW480 cells and interestingly, the SW620 were less migratory. MMP-9 expression increased when the SW480M9 cells were grown on HA and collagen. In addition, SW480M9 cells cultured in the presence of a CD44 activating antibody resulted in a time dependent increase in MMP-9 activity and increased invasion. The SW480M9 cells were shown to be the most adherent to plastic, HA and fibronectin and the SW480M7 were slightly less adherent. The increased HA adhesion in the SW480M9 correlated with increased 80kDa and 120kDa CD44 expression. cDNA microarray analysis was used to identify additional metastasis associated genes by examining and comparing differential gene expression profiles in these cell lines Analysis of gene expression data revealed that 233 genes were significantly up regulated and 208 genes were down regulated in the SW620 when compared to the SW480. Over expression of MMP-7 and MMP-9 in the SW480 cells also caused differential expression. Following literature searches on the gene lists, 5 genes of biological interest were chosen for further analysis and validation. Selected genes were the cysteine protease, cathepsin C, caveolin 1, the nuclear receptor, peroxisome proliferative activated protein gamma (PPARy), the ECM component tenascin C and asparagine synthetase. These clones were sequenced from the cDNA library to verify their identities and real time PCR analysis confirmed the fold changes in expression. Using a fluorescent assay, enhanced cathepsin C enzymatic activity in the SW620 cells was confirmed. A PCR-based method was also used to monitor the expression profiles of particular alternatively spliced tenascin C isoforms in the cell lines studied. An initial pilot study was conducted to examine expression of these genes in matched samples of normal and tumour colon tissue. RNA was extracted from tissues of varying Dukes’ classification and expression was examined by real time PCR. This study identified potential new biomarkers of colorectal metastasis, and examined the involvement of many key players in this process. This study demonstrated that interactions between CD44 and MMPs were important in controlling tumour cell invasion. It is hoped that this work will help elucidate the exact mechanisms of such interactions as well as highlighting their importance in the metastatic process. Further study on these biomarkers and interactions would validate their use as novel markers and therapeutic targets in metastatic colorectal disease.

20069 Background: Some studies suggest a causal relationship between the early placenta insuline-like peptide encoded by the insulin-like 4 gene and cancer invasiveness. Methods: Serum concentrations of pro-EPIL were measured by ELISA using a novel two-site “sandwitch” assay, based on two monoclonal antibodies (mAb) raised against synthetic peptides analogous to two distinct regions of the pro-Epil polypeptide: mAb EPIL15, and biotinylated mAb EPIL02, used as the labelled indicator. The first mAb EPIL15 was purified and bound to a solid phase. Pro-EPIL was allowed to bind, and unbound proteins were removed by washing. The sensitivity limit of the assay was 3 ng/mL of synthetic peptide, and detection was linear over a range of at 3 pg to 200 ng of the synthetic peptide. As primary endpoint, we studied the Pro-EPIL distribution according to some clinico-biological prognostic factors. Results: Between July 2001 and April 2005, nineteen chemonaive NSCLC pts were treated by chemotherapy in our center and serum determinations of pro-EPIL were performed. Ten pts (63%) overexpressed pro-EPIL before start of chemotherapy with a median value of 1.06 ng/mL (range 0.01–6.75). Median age was 60 years (range 38–80), more than 90% of pts were men with an advanced or metastatic disease. Forty-two percent of pts had a large cell carcinoma subtype, with 25% of adenocarcinoma and 25% of epidermoid type. Immunologic evaluations were performed with a median value of the CD3+ lymphocites of 1290/mm3 (range 610–2138). Significant correlations were observed between age, serum alcaline phosphatase and CD3+ number. A border-line positive correlation was observed between pro-EPIL and CD3+ levels (P = 0.09, R2 = 0.5). Median OS for entire cohort was 7.5 months (95% CI, 5.5–9.4), with 67% deaths. Because our sample size was very low, no survival analysis were performed according to the baseline pro-EPIL value and other prognostic factors. A bootstrapping procedure is planned on our data. Conclusions: This results showed that pro-EPIL was overexpressed in the majority of advanced NSCLC pts. Interactions between this biomarker and immune system are possible. Furthemore, pro-EPIL as a therapeutic target might be tested in prospective studies. No significant financial relationships to disclose.

MOST investigations of the invasive process of cancer have involved microscopic studies of tumour spread after injection of cancer cells into experimental animals1–4. Attempts to investigate tumour invasiveness using organ or parabiotic cultures have met with limited success5–7. We have evaluated tumour-mediated cytolysis of normal rat marrow cells using suspension cultures containing Walker-256 carcinosarcoma cells and 59Fe-labelled rat bone marrow cells. This mixed cell culture system seems to provide a promising approach to the further understanding of cancer invasiveness.

In a case-note review of 120 women and 227 men presenting with muscle-invasive bladder tumours in 1998, survival was worse for women in 3 years of follow-up, with the greatest difference, of 19.9%, at 6 months. For more deprived women, 6-month survival was 52.3%, and 32 (37.2%) presented with advanced disease, compared with 73.5%, and three (8.8%) for less deprived women.

Background Despite constant progress, cancer remains the second leading cause of death in the United States. The ability of tumors to metastasize is central to this dilemma, as many studies demonstrate successful treatment correlating to diagnosis prior to cancer spread. Hence a better understanding of cancer invasiveness and metastasis could provide critical insight. Presentation of the hypothesis We hypothesize that a systems biology-based comparison of cancer invasiveness and suburban sprawl will reveal similarities that are instructive. Testing the hypothesis We compare the structure and behavior of invasive cancer to suburban sprawl development. While these two systems differ vastly in dimension, they appear to adhere to scale-invariant laws consistent with invasive behavior in general. We demonstrate that cancer and sprawl have striking similarities in their natural history, initiating factors, patterns of invasion, vessel distribution and even methods of causing death. Implications of the hypothesis We propose that metastatic cancer and suburban sprawl provide striking analogs in invasive behavior, to the extent that conclusions from one system could be predictive of behavior in the other. We suggest ways in which this model could be used to advance our understanding of cancer biology and treatment.