Chick Embryo Chorioallantoic Membrane Research Articles

Preparation, Preliminary Characterization and Antiangiogenic Activities of Polysaccharides from Pomegranate Peels

In the present study, the crude polysaccharides from pomegranate peels (CPP) were prepared by compound enzyme-extraction technology, and the response surface methodology was used to optimize the extraction parameters. Box–Behnken design (BBD) was applied to estimate the effects of extraction temperature, extraction pH, and dosage of compound enzyme on the yield of CPP. A mathematical model with high fitness was obtained. Extraction temperature, extraction pH, and dosage of compound enzyme exhibited independent and interactive effects on CPP yield. CPP were purified by macroporous resin HP-20 and Sephadex G-100 column chromatography to afford purified fraction of CPP-2. The relative molecular weight of CPP-2 was 93.5 kDa. In CPP-2, and FT-IR spectra showed that the main components among CPP-2 may be pectic polysaccharides.The effect of CPP-2 on angiogenesis was measured in vivo by using the chick embryo chorioallantoic membrane (CAM) assay. The results demonstrated that CPP-2 suppressed angiogenesis in chicken embryos.

Curcumin-loaded Polyethyleneimine and chitosan polymer-based Mucoadhesive liquid crystalline systems as a potential platform in the treatment of cervical Cancer

Cervical cancer is the fourth most common cancer in women. Conventional treatments involve surgery, radiotherapy and radiochemotherapy, which affect negatively the life quality. Bioactive molecules, such as curcumin (CUR), are potential alternatives for cervical cancer treatment. However, CUR has limited solubility in water, thus it is interesting to develop drug delivery systems to improve its clinical use. Here, we propose a liquid crystalline system (LCS) as a medium for the formulation and vaginal administration of CUR. To the best of our knowledge, this is the first study that showed the effect of vaginal mucus on the behavior of the liquid crystalline system as well as the antiangiogenic effect of curcumin incorporated in this system in the CAM model. We formulated LCS based on oleic acid, ethoxylated and propoxylated cetyl alcohol and aqueous phase containing water (FW) or dispersion of chitosan associated with polyethyleneimine (FPC). The characterization methodology revealed that LCS presented a lamellar mesophase, which remained stable in the presence of CUR and artificial vaginal mucus. Rheological, texture, and mucoadhesive tests showed that FPC demonstrated properties desirable for vaginal formulations. In vitro release and ex vivo permeation and retention validated the controlled-release properties of the system. LCS released approximately twice as much CUR as the control in 12 h, while ex-vivo porcine vaginal mucosa retained about 2.3% of CUR from LCS. These formulations revealed biocompatibility for L929 cells and toxicity for Hela cells. The vascular changes in response to this potential treatment were evaluated using the chick embryo chorioallantoic membrane (CAM) as a model. The caliber and the number of vessels reduced after the application of CUR-loaded formulations, encouraging the application of the mucoadhesive LCS developed for cervical cancer treatment.

Monocyte Chemoattractant Protein 1 Promotes VEGF-A Expression in OSCC by Activating ILK and MEK1/2 Signaling and Downregulating miR-29c.

Oral squamous cell carcinoma (OSCC) is an aggressive tumor that has a poor prognosis, with high levels of local invasion and lymph node metastasis. Vascular endothelial growth factor A (VEGF-A) plays essential roles in OSCC tumor angiogenesis and metastasis. Monocyte chemoattractant protein-1 (MCP-1, CCL2) is implicated in various inflammatory conditions and pathological processes, including oral cancer. The existing evidence has failed to confirm any correlation between MCP-1 or VEGF-A expression and OSCC angiogenesis. In this study, high expression levels of MCP-1 and VEGF-A were positively correlated with disease stage in patients with OSCC. In oral cancer cells, MCP-1 increased VEGF-A expression and subsequently promoted angiogenesis; miR-29c mimic reversed MCP-1 activity. We also found that MCP-1 modulated VEGF-A expression and angiogenesis through CCR2/ILK/MEK1/2 signaling. Ex vivo results of the chick embryo chorioallantoic membrane (CAM) assay revealed the angiogenic qualities of MCP-1, with increased numbers of visible blood vessel branches. Our data suggest that MCP-1 is a new molecular therapeutic target for the inhibition of angiogenesis and metastasis in OSCC.

Multifunction Sr doped microporous coating on pure magnesium of antibacterial, osteogenic and angiogenic activities

Strontium (Sr) ions were added to porous magnesium (Mg) oxide with silicon and fluorine by microarc oxidation (MAO) to improve its osteogenic and pro-angiogenic properties. First, pure Mg was oxidized by MAO, and Sr was added by electrolysis. The surface of the resulting Sr coating was characterized by SEM, EDS, and EDS mapping. The release of Sr ions was monitored by ICP-OES. The antibacterial property of the coating was assessed against Staphylococcus aureus. The effect of Sr coating on osteogenesis was tested in MC3T3-E1 cell line by performing cell adhesion and proliferation tests, alkaline phosphatase (ALP) activity detection, cell morphology characterization, alizarin red staining, and osteogenic-related gene expression analysis. Finally, HUVECs cells were used to test the effect of Sr coating on angiogenesis through cell migration and tube formation assays, VEGF quantification, chicken embryo chorioallantoic membranes (CAM) test, and angiogenic-related gene expression analysis. The results showed that Sr coating had a hierarchical microstructure with a microporous structure evenly covered with nano-grains and that the Sr elements from the coating were released slowly and continuously. Sr coating had effective antibacterial properties and promoted cell adhesion, proliferation, ALP release, calcium nodule formation, and upregulated osteogenic gene expression. Moreover, the coating could promote migration, tube formation, VEGF expression, and angiogenic gene upregulation in endothelial cells. Sr coating also enhanced angiogenesis of CAM. This study supports that Sr coating on Mg- MAO enhances osteogenesis and angiogenesis.

In Situ DNA/Protein Interaction Assay to Visualize Transcriptional Factor Activation.

The chick embryo chorioallantoic membrane (CAM) represents a powerful in vivo model to study several physiological and pathological processes including inflammation and tumor progression. Nevertheless, the possibility of deepening the molecular processes in the CAM system is biased by the absence/scarcity of chemical and biological reagents, designed explicitly for avian species. This is particularly true for transcriptional factors, proteinaceous molecules that regulate various cellular responses, including proliferation, survival, and differentiation. Here, we propose a detailed antibody-independent protocol to visualize the activation and nuclear translocation of transcriptional factors in cells or in tissues of different animal species. As a proof of concept, DNA/cAMP response element-binding protein (CREB) interaction was characterized on the CAM tissue using oligonucleotides containing the palindromic binding sequence of CREB. Scrambled oligonucleotides were used as controls. In situ DNA/protein interaction protocol is a versatile method that is useful for the study of transcription factors in the cell and tissue of different origins.

Surface functionalization of zeolite-based drug delivery systems enhances their antitumoral activity in vivo

Zeolites have attractive features making them suitable carriers for drug delivery systems (DDS). As such, we loaded the anticancer drug 5-fluorouracil (5-FU), into two different zeolite structures, faujasite (NaY) and Linde Type L (LTL), to obtain different DDS. The prepared DDS were tested in vitro using breast cancer, colorectal carcinoma, and melanoma cell lines and in vivo using the chick embryo chorioallantoic membrane model (CAM). Both assays showed the best results for the Hs578T breast cancer cells, with a higher potentiation for 5-FU encapsulated in the zeolite LTL. To unveil the endocytic mechanisms involved in the internalization of the zeolite nanoparticles, endocytosis was inhibited pharmacologically in breast cancer and epithelial mammary human cells. The results suggest that a caveolin-mediated process was responsible for the internalized zeolite nanoparticles. Aiming to boost the DDS efficacy, the disc-shaped zeolite LTL outer surface was functionalized using amino (NH2) or carboxylic acid (COOH) groups and coated with poly-l-lysine (PLL). Positively functionalized surface LTL nanoparticles revealed to be non-toxic to human cells and, importantly, their internalization was faster and led to a higher tumor reduction in vivo. Overall, our results provide further insights into the mechanisms of interaction between zeolite-based DDS and cancer cells, and pave the way for future studies aiming to improve DDS anticancer activity.

The antibacterial and angiogenic effect of magnesium oxide in a hydroxyapatite bone substitute

Bone graft infections are serious complications in orthopaedics and the growing resistance to antibiotics is increasing the need for antibacterial strategies. The use of magnesium oxide (MgO) is an interesting alternative since it possesses broad-spectrum antibacterial activity. Additionally, magnesium ions also play a role in bone regeneration, which makes MgO more appealing than other metal oxides. Therefore, a bone substitute composed of hydroxyapatite and MgO (HAp/MgO) spherical granules was developed using different sintering heat-treatment cycles to optimize its features. Depending on the sintering temperature, HAp/MgO spherical granules exhibited distinct surface topographies, mechanical strength and degradation profiles, that influenced the in vitro antibacterial activity and cytocompatibility. A proper balance between antibacterial activity and cytocompatibility was achieved with HAp/MgO spherical granules sintered at 1100 ºC. The presence of MgO in these granules was able to significantly reduce bacterial proliferation and simultaneously provide a suitable environment for osteoblasts growth. The angiogenic and inflammation potentials were also assessed using the in vivo chicken embryo chorioallantoic membrane (CAM) model and the spherical granules containing MgO stimulated angiogenesis without increasing inflammation. The outcomes of this study evidence a dual effect of MgO for bone regenerative applications making this material a promising antibacterial bone substitute.

P.222 Brain functional correlates of emotional face processing in body dysmorphic disorder

Strontium (Sr) ions were added to porous magnesium (Mg) oxide with silicon and fluorine by microarc oxidation (MAO) to improve its osteogenic and pro-angiogenic properties. First, pure Mg was oxidized by MAO, and Sr was added by electrolysis. The surface of the resulting Sr coating was characterized by SEM, EDS, and EDS mapping. The release of Sr ions was monitored by ICP-OES. The antibacterial property of the coating was assessed against Staphylococcus aureus. The effect of Sr coating on osteogenesis was tested in MC3T3-E1 cell line by performing cell adhesion and proliferation tests, alkaline phosphatase (ALP) activity detection, cell morphology characterization, alizarin red staining, and osteogenic-related gene expression analysis. Finally, HUVECs cells were used to test the effect of Sr coating on angiogenesis through cell migration and tube formation assays, VEGF quantification, chicken embryo chorioallantoic membranes (CAM) test, and angiogenic-related gene expression analysis. The results showed that Sr coating had a hierarchical microstructure with a microporous structure evenly covered with nano-grains and that the Sr elements from the coating were released slowly and continuously. Sr coating had effective antibacterial properties and promoted cell adhesion, proliferation, ALP release, calcium nodule formation, and upregulated osteogenic gene expression. Moreover, the coating could promote migration, tube formation, VEGF expression, and angiogenic gene upregulation in endothelial cells. Sr coating also enhanced angiogenesis of CAM. This study supports that Sr coating on Mg- MAO enhances osteogenesis and angiogenesis.

Influence of Human Platelet Lysate on Extracellular Matrix Deposition and Cellular Characteristics in Adipose-Derived Stem Cell Sheets.

Adipose-derived stem cell (ASC) is a valuable source of cell therapy. By stimulating extracellular matrix (ECM) secretion, ASC sheets can be fabricated with enhanced regenerative capabilities. In recent years, human platelet lysate (HPL) provides an attractive alternative to fetal bovine serum (FBS) for the ex vivo expansion of ASCs for clinical use. However, the effect of HPL on ASC sheet formation has not been previously determined. In this study, we compared ECM composition and cellular characteristics of ASC sheets cultured in growth medium supplemented with either FBS or HPL. HPL supplement significantly enhanced ASC proliferation without obvious change in the expression pattern of cell surface markers. We found that culturing ASCs with HPL rendered thicker cell sheets with significantly more ECM deposition, including collagen and fibronectin. Proteomic analysis of the FBS or HPL-cultured cell sheets showed diversity in ECM composition. HPL-cultured ASC sheets exhibited up-regulation of interleukin-6 and an anti-inflammatory cytokine, C1q/tumor necrosis factor-related protein-3. Conditioned medium of HPL-cultured ASC sheets significantly enhanced fibroblast migration and tube formation of endothelial cells in vitro, while it inhibited the migration of macrophages toward stimulated macrophages in vitro. TGF-β1-stimulated fibroblasts cultured in ASC sheet-conditioned medium showed down-regulation of α-SMA and TGF-β1. By adding an anti-hepatocyte growth factor (HGF) neutralizing antibody in conditioned medium, we indicated that an anti-fibrosis effect of HPL-cultured ASC sheets is partially mediated through the increased secretion of HGF. Moreover, chick embryo chorioallantoic membrane (CAM) assay showed comparable capillary density after applying either FBS or HPL-cultured ASC sheets, both of which were significantly higher than the control. In conclusion, robust ECM formation with altered ECM composition was noted in ASC sheets cultured in HPL-supplemented medium. Their immunomodulatory and pro-angiogenesis capabilities were largely maintained. Our findings paved the way to elucidate the potential of HPL-cultured ASC sheets for clinical application in tissue regeneration.

Exploitation of the chick embryo chorioallantoic membrane (CAM) as a platform for anti-metastatic drug testing

The establishment of clinically relevant models for tumor metastasis and drug testing is a major challenge in cancer research. Here we report a physiologically relevant assay enabling quantitative analysis of metastatic capacity of tumor cells following implantation into the chorioallantoic membrane (CAM). Engraftment of as few as 103 non-small cell lung cancer (NSCLC) and prostate cancer (PCa) cell lines was sufficient for both primary tumor and metastasis formation. Standard 2D-imaging as well as 3D optical tomography imaging were used for the detection of fluorescent metastatic foci in the chick embryo. H2228- and H1975-initiated metastases were confirmed by genomic analysis. We quantified the inhibitory effect of docetaxel on LNCaP, and that of cisplatin on A549- and H1299-initiated metastatic growths.The CAM assay also mimicked the sensitivity of ALK-rearranged H2228 and EGFR-mutated H1975 NSCLC cells to tyrosine kinase inhibitors crizotinib and gefitinib respectively, as well as sensitivity of LNCaP cells to androgen-dependent enzalutamide therapy. The assay was suggested to reconstitute the bone metastatic tropism of PCa cells. We show that the CAM chick embryo model may be a powerful preclinical platform for testing and targeting of the metastatic capacity of cancer cells.

Inhibition of phospholipase D2 augments histone deacetylase inhibitor-induced cell death in breast cancer cells

BackgroundHistone deacetylase (HDAC) inhibitors are promising anticancer drugs but their effect on tumor treatment has been disappointing mainly due to the acquisition of HDAC inhibitor resistance. However, the mechanisms underlying such resistance remain unclear.MethodsIn this study, we performed Western blot, q-PCR, and promoter assay to examine the expression of HDAC inhibitor-induced phospholipase D2 (PLD2) in MDA-MB231and MDA-MB435 breast cancer cells. Apoptosis and proliferation were analyzed by flow cytometry. In addition to invasion and migration assay, angiogenesis was further measured using in vitro tube formation and chick embryo chorioallantoic membrane model.ResultsHDAC inhibitors including suberoylanilide hydroxamic acid (SAHA), trichostatin, and apicidin, induce expression of PLD2 in a transcriptional level. SAHA upregulates expression of PLD2 via protein kinase C-ζ in breast cancer cells and increases the enzymatic activity of PLD. The combination treatment of SAHA with PLD2 inhibitor significantly enhances cell death in breast cancer cells. Phosphatidic acid, a product of PLD activity, prevented apoptosis promoted by cotreatment with SAHA and PLD2 inhibitor, suggesting that SAHA-induced PLD2 expression and subsequent activation of PLD2 might confers resistance of breast cancer cells to HDAC inhibitor. The combinational treatment of the drugs significantly suppressed invasion, migration, and angiogenesis, compared with that of either treatment.ConclusionThese findings provide further insight into elucidating the advantages of combination therapy with HDAC and PLD2 inhibitors over single-agent strategies for the treatment of cancer.

Angiogenesis-Inflammation Cross Talk in Diabetic Retinopathy: Novel Insights From the Chick Embryo Chorioallantoic Membrane/Human Vitreous Platform.

Pathological angiogenesis of the retina is a key component of irreversible causes of blindness, as observed in proliferative diabetic retinopathy (PDR). The pathogenesis of PDR is complex and involves vascular, inflammatory, and neuronal mechanisms. Several structural and molecular alterations associated to PDR are related to the presence of inflammation that appears to play a non-redundant role in the neovascular response that characterizes the retina of PDR patients. Vascular endothelial growth factor (VEGF) blockers have evolved over time for the treatment of retinal neovascularization. However, several limitations to anti-VEGF interventions exist. Indeed, the production of other angiogenic factors and pro-inflammatory mediators may nullify and/or cause resistance to anti-VEGF therapies. Thus, appropriate experimental models are crucial for dissecting the mechanisms leading to retinal neovascularization and for the discovery of more efficacious anti-angiogenic/anti-inflammatory therapies for PDR patients. This review focuses on the tight cross talk between angiogenesis and inflammation during PDR and describe how the chick embryo chorioallantoic membrane (CAM) assay may represent a cost-effective and rapid in vivo tool for the study of the relationship between neovascular and inflammatory responses elicited by the vitreous humor of PDR patients and for the screening of novel therapeutic agents.

An Experimental Model of Human Recurrent Respiratory Papillomatosis: A Bridge to Clinical Insights.

To investigate the growth pattern of recurrent respiratory papilloma (RRP) implants on chicken embryo chorioallantoic membranes (CAMs) and to evaluate possible associations between the clinical course of the disease and the behavior of experimentally implanted RRP tumors. Experimental study. Fresh 172 RRP tissue samples from 12 patients were implanted onto chick embryo CAMs. Morphological and morphometric analysis of the experimental CAM and chorionic epithelium was performed. The microvascular network of the CAM with the RRP implant was investigated under the effect of fluoresceinated anionic dextran. The peculiarities of the clinical course of the disease were evaluated. The implanted RRP tissue samples survived on CAMs in 86% of cases, retaining their essential morphologic characteristics and proliferative capacity of the original tumor. Implants induced thickening of both the CAM and the chorionic epithelium, but none of the RRP implants crossed the basement membrane of the hosting CAM. A "crawling film" of acellular material with newly formed papilloma sprouts located on the outer chorionic epithelium of the CAM was detected. Direct association between a recurrence rate of RRP and the number of newly formed papilloma sprouts around the implanted tumor on CAMs was revealed. The chicken embryo CAM-based model is appropriate for investigations of RRP and facilitates the understanding of tumor biology and the clinical course of the disease, thus providing the basis for further research and acceleration of the identification and development of new specific therapeutic compounds that limit the spread and recurrence of RRP. N/A Laryngoscope, 131:E914-E920, 2021.

Lactobacillus acidophilus and Bifidobacterium longum exhibit antiproliferation, anti-angiogenesis of gastric and bladder cancer: Impact of COX2 inhibition

BackgroundProbiotics have gained exponential attention worldwide because they are used as a therapeutic tool for management, prevention and treatment of diseases like cancer. Here, antiproliferation, anti-angiogenesis and antitumor effects of Lactobacillus acidophilus (LA) and Bifidobacterium longum (BL) were examined on gastric (AGS) and bladder (J253) cancer cell lines. MethodsCancer cell viability was monitored with MTT assay. The chick embryo chorioallantoic membrane (CAM) assay was used to screen tumor growth and hemoglobin content of cancer cells alone and with different LA and BL concentrations. Protein expression of COX2 was analyzed with Western blot in cell lines treated with LA, BL, celecoxib at 60 and 100 μM, or their combination. ResultsTreatment with either probiotic showed increased vacuolated gastric cancer (AGS) cells with significant morphological changes at high concentration (2.5 × 107 cfu/ml) of LA and BL bacteria compared to bladder cancer cells (J253). Similarly, AGS cells responded well to LA and BL and there was decreased cancer cell viability, tumor weight and hemoglobin content (p < 0.001). The probiotics were more potent against AGS than J253. High dose (2.5 × 107 cfu/ml) of probiotic bacteria when combined with 60 μM celecoxib downregulated COX2 expression in AGS by 70 % and 95 % (p < 0.001), without affecting COX1 expression. However, J253 cells did not show any changes in the expression of COX2 with probiotics, which is consistent with no effect on cancer cell viability and tumor angiogenesis. ConclusionsThis study is the first to document antiproliferation and anti-angiogenesis of LA and BL against gastric cancer by downregulating COX2 expression. LA and BL might be new, safe agents for targeting COX2 and suppressing growth of some cancers.

Characterization of markers of cold-adapted candidate virus strains for live attenuated vaccines against chickenpox and shingles

Introduction. Chickenpox poses a significant public health concern due to its worldwide occurrence, a substantial probability of severe clinical progression, development of complications that can lead to a fatal outcome. Routine vaccination is the only way to prevent the disease. The purpose of this study was to assess the attenuation of cold-adapted (CA) candidate virus strains of Varicella zoster and Herpes zoster by using traditional and new methods.Materials and methods. The study was performed on strains of diploid cells from human embryonic lung and musculocutaneous tissue, primary and diploid cells of guinea pig fetal fibroblasts. Two clinical isolates of the virus were obtained — from a child with chickenpox and from an adult during the reactivation of shingles. The vOka vaccine strain and Ellen strain, a laboratory strain, were used as a control. The viral infectivity was measured by using a sensitive limiting dilution assay. The virulence was measured through the analysis of chick embryo chorioallantoic membranes infected with the Varicella zoster virus.Results. The clinical isolates were sub-cultured at lower temperatures, put through comparative tests and checked for presence of attenuation biomarkers. It was found that vFiraVax, a Varicella zoster virus strain, and vZelVax, a Herpes zoster virus strain were temperature-sensitive and cold-adaptable, but they lacked virulence. Attenuated CA virus strains induced lower expression of IFN-α and IFN-γ receptors on human mononuclear cells as compared to their parental variants.Conclusion. We created and assessed two candidate vaccine strains through attenuation of clinical isolates.

Biological characteristics of endometriotic mesenchymal stem cells isolated from ectopic lesions of patients with endometriosis

BackgroundResearch into the pathogenesis of endometriosis (EMs) would substantially promote its effective treatment and early diagnosis. However, the aetiology of EMs is poorly understood and controversial despite the progress in EMs research in the last several decades. Currently, accumulating evidence has shed light on the importance of endometrial stem cells (EnSCs) residing in the basal layer of endometrium in the establishment and progression of endometriotic lesions. Therefore, we aimed to identify the differences between EnSCs isolated from the ectopic lesions of EMs patients (EnSC-EM-EC) and EnSCs isolated from eutopic endometrium of control group (EnSC-Control). We further performed preliminary exploration of the potential signalling pathways involved in the above abnormalities.MethodsEnSC-EM-EC (n = 12) and EnSC-Control (n = 13) were successfully isolated. Then, the proliferative capacity, migratory capacity and angiogenic potential of EnSCs were evaluated by conventional MTT assay, flow cytometry, wound healing assay, transwell assay, tube formation assay and chick embryo chorioallantoic membrane assay respectively. The expression of 11 angiogenesis-associated biological factors and 11 cytokines secreted by EnSCs and 17 adhesion molecules expressed on EnSCs were determined by protein array assays respectively. Differentially expressed genes (DEGs) between EnSC-EM-EC and EnSC-Control were analysed by RNA-sequence.ResultsEnSC-EM-EC exhibited unique biological characteristics, including prolonged mitosis, enhanced migratory capacity and enhanced angiogenic potential. Greater amounts of angiogenic factors (especially VEGF and PDGF) were secreted by EnSC-EM-EC than by EnSC-Control; however, the distinct profiles of cytokines secreted by EnSC-EM-EC and adhesion molecules expressed by EnSC-EM-EC require further investigation. A total of 523 DEGs between EnSC-EM-EC and EnSC-Control were identified and analysed using the KEGG and Gene Ontology databases.ConclusionsOur results not only improve the understanding of EMs but also contribute to the development of EnSC-EM-EC as a tool for EMs drug discovery. These cells could be of great help in exploiting promising therapeutic targets and new biomarkers for EMs treatment and prognosis.

A Bromodomain-Containing Protein 4 (BRD4) Inhibitor Suppresses Angiogenesis by Regulating AP-1 Expression.

Angiogenesis dysregulation contributes to inflammation, infections, immune disorders, and carcinogenesis. Bromodomain-containing protein 4 (BRD4) is an epigenetic reader that recognizes histone proteins and acts as a transcriptional regulator to trigger tumor growth and the inflammatory response. The pan-bromodomain and extra-terminal domain (BET) inhibitor, (+)-JQ1 (1), was reported to inhibit angiogenesis. However, owing to the non-selectivity action of (+)-JQ1 towards all BET family members, the role of BRD4 and that of its bromodomains (BD1 and BD2) in angiogenesis remains elusive. Herein, we identified a potent BRD4 inhibitor, ZL0513 (7), which exhibited significant anti-angiogenic effects in chick embryo chorioallantoic membrane (CAM) and yolk sac membrane (YSM) models. This inhibitor also directly suppressed the viability and tube formation of human umbilical vascular endothelial cells (HUVECs). Moreover, ZL0513 (7) was found to inhibit the phosphorylation of c-jun and c-fos, important members of activating protein-1 (AP-1) transcription factor complexes that enhance angiogenesis. The findings on this novel BRD4 inhibitor indicate that, in addition to being a powerful pharmacological tool for further elucidating the roles and functions of BRD4 and its BD domains in angiogenesis, it may serve as a potential therapeutic strategy for targeting the vasculature in various angiogenesis-dysregulated human diseases.

Targeting lactate production and efflux in prostate cancer

Prostate cancer (PCa) is the most commonly diagnosed cancer in men worldwide. Screening and management of PCa remain controversial and, therefore, the discovery of novel molecular biomarkers is urgently needed. Alteration in cancer cell metabolism is a recognized hallmark of cancer, whereby cancer cells exhibit high glycolytic rates with subsequent lactate production, regardless of oxygen availability. To maintain the hyperglycolytic phenotype, cancer cells efficiently export lactate through the monocarboxylate transporters MCT1 and MCT4. The impact of inhibiting lactate production/extrusion on PCa cell survival and aggressiveness was investigated in vitro and ex vivo using primary tumor and metastatic PCa cell lines and the chicken embryo chorioallantoic membrane (CAM) model. In this study, we showed the metastatic PCa cell line (DU125) displayed higher expression levels of MCT1/4 isoforms and glycolysis-related markers than the localized prostate tumor-derived cell line (22RV1), indicating these proteins are differentially expressed throughout prostate malignant transformation. Moreover, disruption of lactate export by MCT1/4 silencing resulted in a decrease in PCa cell growth and motility. To support these results, we pharmacological inhibited lactate production (via inhibition of LDH) and release (via inhibition of MCTs) and a reduction in cancer cell growth in vitro and in vivo was observed. In summary, our data provide evidence that MCT1 and MCT4 are important players in prostate neoplastic progression and that inhibition of lactate production/export can be explored as a strategy for PCa treatment.

Discovery of new fluorescent thiazole\u2013pyrazoline derivatives as autophagy inducers by inhibiting mTOR activity in A549 human lung cancer cells

A series of fluorescent thiazole–pyrazoline derivatives was synthesized and their structures were characterized by 1H NMR, 13C NMR, and HRMS. Biological evaluation demonstrated that these compounds could effectively inhibit the growth of human non-small cell lung cancer (NSCLC) A549 cells in a dose- and time-dependent manner in vitro and inhibit tumor growth in vivo. The structure–activity relationship (SAR) of the compounds was analyzed. Further mechanism research revealed they could induce autophagy and cell cycle arrest while had no influence on cell necrosis. Compound 5e inhibited the activity of mTOR via FKBP12, which could be reversed by 3BDO, an mTOR activator and autophagy inhibitor. Compound 5e inhibited growth, promoted autophagy of A549 cells in vivo. Moreover, compound 5e showed good selectivity with no influence on normal vascular endothelial cell growth and the normal chick embryo chorioallantoic membrane (CAM) capillary formation. Therefore, our research provides potential lead compounds for the development of new anticancer drugs against human lung cancer.

Chick embryo chorioallantoic membrane as a suitable in vivo model to evaluate drug delivery systems for cancer treatment: A review

Cancer represents a significant public health problem. More than 18.1 million people are annually diagnosed with cancer and 9.6 million die mainly due to metastatic disease. Chemotherapy has been one of the main cancer treatment modalities; however, most of the chemotherapeutic agents are non-specific, exhibiting several toxic side effects, which compromises the patient’s quality of life. Therefore, it is necessary to search for new therapeutic alternatives, using for example, drug delivery systems (DDS) to target cancer cells, increasing the selectivity of chemotherapeutic drugs. This approach is promising; however, it is crucial to evaluate the biological performance of the systems. Although mammalian models continue to be explored for clinical applications, they are time-consuming and very restrictive from the ethical and legal perspectives. Hence, the chick embryo chorioallantoic membrane (CAM) has been shown to be a suitable in vivo model since it allows a more appropriate model for the study of drugs and/or DDS performance than in vitro tests. Thereby, this article revises the recent advances of DDS for cancer therapy, evaluating the feasibility of the CAM model.