Human Glioblastoma Cancer Research Articles

Targeting BMI-1 with PLGA\u2013PEG nanoparticle-containing PTC209 modulates the behavior of human glioblastoma stem cells and cancer cells

Despite advances in glioblastoma (GBM) treatments, current approaches have failed to improve the overall survival of patients. The oncogene BMI-1, a core member of the polycomb group proteins, is a potential novel therapeutic target for GBM. To enhance the efficacy and reduce the toxicity, PTC209, a BMI-1 inhibitor, was loaded into a PLGA–PEG nanoparticle conjugated with CD133 antibody (Nano-PTC209) and its effect on the behavior of human GBM stem-like cells (GSCs) and the human glioblastoma cell line (U87MG) was assessed. Nano-PTC209 has a diameter of ~ 75 nm with efficient drug loading and controlled release. The IC50 values of Nano-PTC209 for GSCs and U87MG cells were considerably lower than PTC209. Nano-PTC209 significantly decreased the viability of both GSCs and U87MG cells in a dose-dependent manner and caused a significant enhancement of apoptosis and p53 levels as well as inhibition of AKT and JNK signaling pathways. Furthermore, Nano-PTC209 significantly inhibited the migration ability, decreased the activity of metalloproteinase-2 and -9, and increased the generation of reactive oxygen species in both GSCs and U87MG cells. Our data indicate that PLGA–PEG nanoparticle conjugated with CD133 antibody could be an ideal nanocarrier to deliver PTC209 and effectively target BMI-1 for potential approaches in the treatment of GBM.

Gold Nanoparticles Synthesized Using Extracts of Cyclopia intermedia, Commonly Known as Honeybush, Amplify the Cytotoxic Effects of Doxorubicin.

Cyclopia intermedia (C. intermedia) is an indigenous South African shrub used to prepare the popular medicinal honeybush (HB) tea. This plant contains high levels of mangiferin (MGF), a xanthonoid that was reported to have numerous biological activities, including anti-tumor activity. MGF and extracts that contain high concentrations of MGF, such as extracts from Mangifera indica L. or mango have been used to synthesize gold nanoparticles (AuNPs) using green nanotechnology. It has previously been shown that when AuNPs synthesized from M. indica L. extracts are used in combination with doxorubicin (DOX) and Ayurvedic medicine, the anti-tumor effects appear to be augmented. It has also been demonstrated that MGF used in combination with DOX resulted in enhanced anti-tumor effects. In this study, C. intermedia (HB) and MGF were used to synthesize HB-AuNPs and MGF-AuNPs, respectively. The physicochemical properties of the AuNPs were characterized by the UV-Visible Spectroscopy (UV-Vis), dynamic light scattering (DLS), Fourier transform infra-red spectroscopy (FTIR), X-ray diffraction spectroscopy (XRD) and high-resolution transmission electron microscopy (HR-TEM). The cytotoxicity of HB-AuNPs and MGF-AuNPs were assessed on human colon (Caco-2), prostate (PC-3) and glioblastoma (U87) cancer cells; as well as normal breast epithelial (MCF-12A) cells using the MTT assay. Both HB-AuNPs and MGF-AuNPs demonstrated relatively low cytotoxicity in these cells. However, when these nanoparticles were used in combination with DOX, the cytotoxicity of DOX was significantly augmented.

Assessment of replication of bovine herpesvirus type 4 in human glioblastoma and breast cancer cells as a potential oncolytic virus.

Oncolytic viruses have been extensively used in cancer treatment due to their tropism, selective replication only in tumor cells, and possible synergic interaction with other therapeutics. Different researchers have demonstrated that bovine herpesvirus 4 (BoHV-4), a member of the gammaherpesviridae family, has oncolytic potential in some human-origin cancer cell lines like glioma through the selective replication strategy. Using four apoptosis detection methods, namely MTT, LDH, TUNEL, and Annexin V assays, we evaluated the apoptotic effect of BoHV-4 Movar33/63 reference strain along with a recombinant BoHV-4 expressing EGFP in U87 MG cells (human glioblastoma cell line), MDA MB-231 (human breast cancer cell line), and MCF10a (non-tumorigenic human mammary epithelial cell line). Our findings indicate that this virus can replicate and induce apoptosis in these cell lines and hinder in vitro proliferation in a dose-dependent manner. In conclusion, BoHV-4 has in vitro potential as a novel oncolytic virus in human cancer therapy. However, its replication potential in the MCF10a cells as a non-tumorigenic human mammary epithelial cell line is a concern in using this virus in cancer therapy, at least against human mammary tumors. Further studies must therefore be conducted to examine the specific apoptotic pathways induced by this virus to move on to further experiments.

Production and characterization of bacterial cellulose scaffold and its modification with hyaluronic acid and gelatin for glioblastoma cell culture

Three-dimensional (3D) in vitro cell culture models have recently gained increasing interest in predicting the response of anticancer drugs. In this study first, we tried to obtain a novel hyaluronic acid (HA)/gelatin (Gel) modified bacterial cellulose (BC) composite scaffolds by in situ fermentation method. Morphological and chemical structures, wettability, and thermal stability of scaffolds were evaluated. In particular, the human glioblastoma (GBM) cancer cell line (U251) was seeded into BC/HA/Gel scaffolds to evaluate their potential as in vitro 3D cancer cell culture. MTT proliferation assay, scanning electron microscopy, and confocal microscopy were utilised to determine cell proliferation, morphology and adhesion. The results suggest that our hyaluronic acid and gelatin modified bacterial cellulose scaffold is promising to be used as in vitro 3D culture of GBM cells and may be used to predict treatment response or reactions of new therapeutics.

Lipophilic dye-compatible brain clearing technique allowing correlative magnetic resonance/high-resolution fluorescence imaging in rat models of glioblastoma

In this work we optimized a novel approach for combining in vivo MRI and ex vivo high-resolution fluorescence microscopy that involves: (i) a method for slicing rat brain tissue into sections with the same thickness and spatial orientation as in in vivo MRI, to better correlate in vivo MRI analyses with ex-vivo imaging via scanning confocal microscope and (ii) an improved clearing protocol compatible with lipophilic dyes that highlight the neurovascular network, to obtain high tissue transparency while preserving tissue staining and morphology with no significant tissue shrinkage or expansion. We applied this methodology in two rat models of glioblastoma (GBM; U87 human glioma cells and patient-derived human glioblastoma cancer stem cells) to demonstrate how vital the information retrieved from the correlation between MRI and confocal images is and to highlight how the increased invasiveness of xenografts derived from cancer stem cells may not be clearly detected by standard in vivo MRI approaches. The protocol studied in this work could be implemented in pre-clinical GBM research to further the development and validation of more predictive and translatable MR imaging protocols that can be used as critical diagnostic and prognostic tools. The development of this protocol is part of the quest for more efficacious treatment approaches for this devastating and still uncurable disease. In particular, this approach could be instrumental in validating novel MRI-based techniques to assess cellular infiltration beyond the macroscopic tumor margins and to quantify neo-angiogenesis.

The antiproliferative potential of isolated emodin and aloe-emodin from Rheum ribes on different cancer cell lines

Herbal compounds are important resources that used in the treatment of many diseases, and are inspiring in drug design and development. Rheum ribes, a perennial native plant, has potentially diverse medicinal effects, including antioxidant, antibacterial and anti-tumor activity. The key molecules associated with anticancer activity of R. ribes in tumor formation still remains unclear. The present study was aimed to elucidate the antiproliferative effects of two purified compounds, emodin and aloe-emodin, from R. ribes. In our study, chromatographic separation was used to purification of compounds. Experimental 1D and 2D 1H and 13C-NMR spectra helped us to confirm the chemical structures of emodin and aloe-emodin. The viability of human glioblastoma cell line (U373), human breast carcinoma cells (MCF-7), and human colorectal cancer cell line (HT-29) were detected by WST-1 proliferation assay as dose- and time-dependent. The isolated emodin and aloe-emodin from R. ribes, reduced viability of human glioblastoma, breast and colorectal cancer cell lines in dose- and time-dependent manner significantly. These antiproliferative effects may contribute as a possible strategy to the development of new therapeutic approaches in the treatment of cancer.

JAM-A functions as a female microglial tumor suppressor in glioblastoma.

Glioblastoma (GBM) is the most aggressive primary brain tumor and has a dismal prognosis. Previously, we identified that junctional adhesion molecule A (JAM-A), a cell adhesion molecule, is highly elevated in human GBM cancer stem cells and predicts poor patient prognosis. While JAM-A is also highly expressed in other cells in the tumor microenvironment, specifically microglia and macrophages, how JAM-A expression in these cells affects tumor growth has yet to be determined. The goal of this study was to understand the role of microenvironmental JAM-A in mediating GBM growth. Male and female wild-type (WT) and JAM-A-deficient mice were transplanted intracranially with the syngeneic glioma cell lines GL261 and SB28 and were assessed for differences in survival and microglial activation in tumors and in vitro. RNA-sequencing was performed to identify differentially regulated genes among all genotypes, and differences were validated in vitro and in vivo. We found that JAM-A-deficient female mice succumbed to GBM more quickly compared with WT females and JAM-A-deficient and male WT mice. Analysis of microglia in the tumors revealed that female JAM-A-deficient microglia were more activated, and RNA-sequencing identified elevated expression of Fizz1 and Ifi202b specifically in JAM-A-deficient female microglia. Our findings suggest that JAM-A functions to suppress pathogenic microglial activation in the female tumor microenvironment, highlighting an emerging role for sex differences in the GBM microenvironment and suggesting that sex differences extend beyond previously reported tumor cell-intrinsic differences.

Synergistic or Additive Pharmacological Interactions between Magnoflorine and Cisplatin in Human Cancer Cells of Different Histological Origin.

Magnoflorine is an aporphine alkaloid present in plant species belonging to the Berberidaceae, Magnoliaceae, Menispermaceae, or Papaveraceae botanical families. The interest of magnoflorine has increased recently due to its multiplicity of pharmacological properties. The aim of this study was the analysis of combined anti-proliferative effect of magnoflorine and cisplatin and the assessment of drug–drug pharmacological interaction between these agents using isobolographic method in MDA-MB-468 human breast, NCIH1299 lung, TE671 rhabdomyosarcoma, or T98G glioblastoma cancer cell lines. Magnoflorine in combination with cisplatin at a fixed ratio of 1:1 augmented their anticancer action and yielded synergistic or additive pharmacological interactions by means of isobolographic method, therefore combined therapy using these two active agents can be a promising chemotherapy regimen in the treatment of some types of breast, lung, rhabdomyosarcoma, and glioblastoma cancers.

Cross Interaction between M2 Muscarinic Receptor and Notch1/EGFR Pathway in Human Glioblastoma Cancer Stem Cells: Effects on Cell Cycle Progression and Survival.

Glioblastomas (GBM) are the most aggressive form of primary brain tumors in humans. A key feature of malignant gliomas is their cellular heterogeneity. In particular, the presence of an undifferentiated cell population of defined Glioblastoma Stem cells (GSCs) was reported. Increased expression of anti-apoptotic and chemo-resistance genes in GCSs subpopulation favors their high resistance to a broad spectrum of drugs. Our previous studies showed the ability of M2 muscarinic receptors to negatively modulate the cell growth in GBM cell lines and in the GSCs. The aim of this study was to better characterize the inhibitory effects of M2 receptors on cell proliferation and survival in GSCs and investigate the molecular mechanisms underlying the M2-mediated cell proliferation arrest and decreased survival. Moreover, we also evaluated the ability of M2 receptors to interfere with Notch1 and EGFR pathways, whose activation promotes GSCs proliferation. Our data demonstrate that M2 receptors activation impairs cell cycle progression and survival in the primary GSC lines analyzed (GB7 and GB8). Moreover, we also demonstrated the ability of M2 receptor to inhibit Notch1 and EGFR expression, highlighting a molecular interaction between M2 receptor and the Notch-1/EGFR pathways also in GSCs.

Tragacanth-mediate synthesis of NiO nanosheets for cytotoxicity and photocatalytic degradation of organic dyes.

In this study, NiO nanosheets have been manufactured using a co-precipitation approach that involved the usage of nickel nitrate (Ni (NO3)2.6H2O) as the raw material and tragacanth in the role of a stabilizing agent. NiO nanosheets have been fabricated through the reduction of nickel nitrate solution that had been obtained by the application of aqueous extract of tragacanth, which is capable of functioning as a reducing and stabilizing agent. In the following, the physical and chemical properties of tragacanth-stabilized NiO nanosheets have been identified via FESEM, EDS, XRD, UV-Vis, and FT-IR techniques. According to the XRD pattern, these particular nanosheets have contained a cubic structure and group space Fm3m, along with the average size of about 18 to 43nm that had been in agreement with the FESEM measurements. In addition, we have evaluated the photocatalytic activity of tragacanth-stabilized NiO nanosheets on the degradations of methylene blue (MB) and methyl orange (MO) dyes. The performed photocatalytic assessment has displayed that the nanosheets can degrade 82% of MO within 210min and 60% of MB in 300min. The cytotoxicity of tragacanth-stabilized NiO nanosheets on human Glioblastoma cancer (U87MG) cell lines has been investigated via the MTT assay, while it has been detected in the obtained results that the inhibitory concentration (IC50) had been 125µg/mL.

TiO2 hollow nanospheres functionalized with folic acid and ZnPc for targeted photodynamic therapy in glioblastoma cancer

Abstract

The Effect of Diopatra claparedii Grube, 1878 Aqueous Extract on Chang Liver and Human Primary Glioblastoma (U-87) Cell Lines

Due to the distinctive regenerative ability of Diopatra claparedii Grube, 1878, the local marine polychaete has the potential as a cellular growth agent. In this study, the growth effect was investigated in normal cells and cancer cells. Different concentrations (0-100mg/mL) of D. claparedii aqueous extract were tested on Chang Liver (normal cells), and Human Primary Glioblastoma (U-87) (cancer cells) cell lines for 24, 48 and 72 hours. Percentage of cell viability was evaluated by [2-(4, 5-dimethyl-2-thiazolyl)-3, 5-diphenyl-2H tetrazolium bromide] (MTT) assay. The findings suggested that the extract had a proliferative effect on normal cell growth when tested at lower doses (<60 mg/mL) but inhibited normal cells at concentrations >80 mg/mL in all incubation periods. Meanwhile, it showed the cytotoxic effect on cancer cells only after 48h when treated with all concentrations. As demonstrated, the extract could induce normal cell growth without causing abnormal or cancer cells progression at low concentrations after 48h and 72h.

TNF-derived peptides inhibit tumour growth and metastasis through cytolytic effects on tumour lymphatics.

Tumour necrosis factor (TNF) is a multi-functional cytokine with profound and diverse effects on physiology and pathology. Identifying the molecular determinants underlying the functions and pathogenic effects of TNF is key to understanding its mechanisms of action and identifying new therapeutic opportunities based on this important molecule. Previously, we showed that some evolutionarily conserved peptides derived from TNF could induce cell death (e.g. apoptosis and/or necrosis), a feature of immune defence mechanisms shared by many vertebrates. In this study, we demonstrated that necrosis-inducing peptide P16 kills human glioblastoma cancer cells and primary human hepatoma or renal cancer cells isolated from patients who had not responded to standard treatments. Importantly, we show that the necrosis-inducing peptide P1516 significantly improves survival by inhibiting tumour metastasis in a 4T1 breast cancer syngeneic graft mouse model. Because the lymphatic system is an important metastatic route in many cancers, we also tested the effect of TNF-derived peptides on monolayers of primary human lymphatic endothelial cells (hDLEC) and found that they increased junctional permeability by inducing cytoskeletal reorganization, gap junction formation and cell death. Transmission electron microscopy imaging evidence, structural analysis and in-vitro liposome leakage experiments strongly suggest that this killing is due to the cytolytic nature of these peptides. P1516 provides another example of a pro-cytotoxic TNF peptide that probably functions as a cryptic necrotic factor released by TNF degradation. Its ability to inhibit tumour metastasis and improve survival may form the basis of a novel approach to cancer therapy.

Abstract LB-110: Proteasome addiction a new therapeutic opportunity to treat PTEN-deficient brain tumors

Abstract One of the most common genetic alterations in glioblastoma (30-40%) occurs in the tumor suppressor gene PTEN (phosphatase and tensin homolog), where loss of function has been mechanistically linked to increased tumor cell invasion, and to a lack of radio- and chemo-therapy response. To identify new drug compounds that target PTEN-deficient brain tumors we performed a high throughput drug screening using patient-derived GBM spheres and found that PTEN-deficient samples were highly sensitive to proteasome inhibitors. We confirmed PTEN dependency to proteasome inhibition by genetically overexpressing or deleting PTEN in GBM cells and established that the drug inhibition response relied on PTEN enzymatic activity. Additionally, proteasome inhibition specifically suppressed tumor growth and increased survival of mice orthotopically engrafted with human PTEN-null glioblastoma cancer stem cells. Mechanistically, we determined that PTEN-deficient cells were more sensitive to proteasome inhibition due to an increase in protein synthesis rate and loss of autophagy activity associated with activation of the PI3K/mTOR pathway. Finally, integrated in silico analysis of the Samsung Medical Center dataset corroborated that samples with PTEN/EGFR/PI3K alterations were significantly more sensitive to Carfilzomib, an FDA-approved proteasome inhibitor. This study reveals that PTEN-deficient cells are "proteasome addictive" and opens a new therapeutic opportunity to treat glioblastoma PTEN-deficient patients. Citation Format: Jorge A. Benitez, Darren Finlay, Alexandre Rosa-Campos, Jianhui Ma, Tomoyuki Koga, Kristiina Vuori, Frank Furnari. Proteasome addiction a new therapeutic opportunity to treat PTEN-deficient brain tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr LB-110.

Application of high-mannose-type glycan-specific lectin from Oscillatoria Agardhii for affinity isolation of tumor-derived extracellular vesicles

Tumor cells secrete membrane vesicles of various sizes, termed extracellular vesicles (EVs), which have gained increasing attention as potential tumor diagnostic markers. Tumor-derived EVs are enriched with high-mannose-type glycans. Here, we report the affinity isolation of EVs from human melanoma A375 cells by using high-mannose-type glycan-specific agglutinin from Oscillatoria Agardhii (OAA). Glycan analysis of melanoma EVs revealed the presence of high-mannose-type glycans with structural units preferred by OAA. We showed that in solution, OAA binds to melanoma EVs in a high-mannose-type glycan-dependent manner. Furthermore, OAA-immobilized beads were found to capture 60% of the particles and most proteinous components from melanoma EVs. Major EV glycoproteins that potentially interact with OAA were identified to be cluster of differentiation 109 (CD109), integrin α6 and a disintegrin and metalloproteinase domain-containing protein 10 (ADAM10). In addition to melanoma EVs, OAA captured EVs from human lung cancer, glioblastoma and colon cancer cells, but not those from endothelial cells and fibroblasts. These results indicate that OAA-immobilized beads may serve as a novel platform for affinity-capture of tumor-derived EVs.

<p>Upregulation of microRNA-1270 suppressed human glioblastoma cancer cell proliferation migration and tumorigenesis by acting through WT1</p>

BackgroundGlioblastoma multiforme (GBM) is one of the most aggressive brain tumors among human beings. In this study, we explored the functions of human microRNA-1270 (hsa-miR-1270) on GBM cancer cell proliferation, migration, and tumorigenesis.Materials and methodsIn GBM cell lines and clinical tissues, hsa-miR-1270 expression was probed by quantitative real-time PCR (qRT-PCR). In LN-18 and A172 cells, hsa-miR-1270 was upregulated by lentiviral transduction. The effects of hsa-miR-1270 upregulation on GBM in vitro and in vivo functions were probed by proliferation, migration, and xenograft assays, respectively. The correlation between hsa-miR-1270 and Wilms’ tumor gene (WT1) was probed by dual-luciferase activity assay, qRT-PCR, and Western blot. WT1 was then secondarily over-expressed in hsa-miR-1270-upregulated LN-18 and A172 cells, to explore its mechanisms in GBM’s association with hsa-miR-1270.ResultsHsa-miR-1270 was significantly downregulated in both GBM cell lines and clinical tumors. Upregulating hsa-miR-1270 considerably suppressed GBM cell proliferation and migration in vitro and xenograft in vivo. WT1 was inversely correlated with hsa-miR-1270 in GBM. WT1 overexpression in hsa-miR-1270-upregulated GBM cells reversed the anticancer functions of hsa-miR-1270 on cancer proliferation and migration.ConclusionHsa-miR-1270 upregulation may have suppressing effects on GBM cancer cells, likely by functionally acting through WT1.

Histone Deacetylase Inhibitors Impair Vasculogenic Mimicry from Glioblastoma Cells.

Glioblastoma (GBM), a high-grade glioma (WHO grade IV), is the most aggressive form of brain cancer. Available treatment options for GBM involve a combination of surgery, radiation and chemotherapy but result in a poor survival outcome. GBM is a high-vascularized tumor and antiangiogenic drugs are widely used in GBM therapy as adjuvants to control abnormal vasculature. Vasculogenic mimicry occurs in GBM as an alternative vascularization mechanism, providing a means whereby GBM can escape anti-angiogenic therapies. Here, using an in vitro tube formation assay on Matrigel®, we evaluated the ability of different histone deacetylase inhibitors (HDACis) to interfere with vasculogenic mimicry. We found that vorinostat (SAHA) and MC1568 inhibit tube formation by rat glioma C6 cells. Moreover, at sublethal doses for GBM cells, SAHA, trichostatin A (TSA), entinostat (MS275), and MC1568 significantly decrease tube formation by U87MG and by patient-derived human GBM cancer stem cells (CSCs). The reduced migration and invasion of HDACis-treated U87 cells, at least in part, may account for the inhibition of tube formation. In conclusion, our results indicate that HDACis are promising candidates for blocking vascular mimicry in GBM.

Low glucose availability alters the expression of genes involved in initial adhesion of human glioblastoma cancer cell line SF767.

Studying the metabolic pathways of cancer cells is considered as a key to control cancer malignancies and open windows for effective drug discovery against cancer. Of all the properties of a tumor, metastasis potential is a defining characteristic. Metastasis is controlled by a variety of factors that directly control the expression of cell adhesion proteins. In this study we have investigated the expression of cell to cell and cell to matrix adhesion protein genes during the initial phases of attachment of human glioblastoma cancer cell line SF767 (66Y old human female: UCSF Neurosurgery Tissue Bank) to the attachment surface under (Cell culture treated polystyrene plate bottom) glucose-rich and glucose-starved conditions. The aim was to imitate the natural microenvironment of glucose availability to cancer cells inside a tumor that triggers epithelial to mesenchymal transition (EMT). In this study, we have observed the gene expression of epithelial and mesenchymal isoforms of cadherin (E-CAD and N-CAD) and Ig like cell adhesion molecules (E-CAM and N-CAM) along with Integrin family subunits for the initial attachment of cancer cells. We observed that high glucose environments promoted cell survival and cell adhesion, whereas low glucose accelerated EMT by downregulating the expression level of integrin, E-CAD, and N-CAD, and upregulation of N-CAM during early period of cell adhesion. Low glucose availability also downregulated variety of structural and regulatory genes, such as zinc finger E-box binding home box 1A), cytokeratin, Snail, and β catenin, and upregulation of hypoxia-inducible factor 1, matrix metalloprotease 13/Collagenase 3, vimentim, p120, and fructose 1,6 bisphosphatase. Glucose conditions are more efficient for cancer studies in this case glioblastoma cells.

Cytotoxicity and apoptogenic properties of the standardized extract of Portulaca oleracea on glioblastoma multiforme cancer cell line (U-87): a mechanistic study.

The traditional uses of Portulaca oleracea L. (PO) with anti-inflammatory and anti-cancer activity as well as antioxidants properties were expressed previously. Glioma is considered the most common primary brain tumor and its malignant form is the most lethal adult brain tumor, that glioblastoma covers about 50 % of glioma tumors. The present study was aimed to evaluate the cytotoxicity and apoptogenic effects of the hydro-ethanolic extract of PO on human glioblastoma cancer cell line (U-87) and the role of NF-κB. Cytotoxicity of the extract in the presence or absence of Vitamin C was evaluated using MTT assay, and the following hypotonic PI and SubG1 peak were performed. Moreover, the reactive oxygen species (ROS), the level of NF-κB protein and nitric oxide (NO) production were investigated. The extract had cytotoxicity and apoptogenic effects on U-87 cells in both the concentration and time-dependent manners. The mechanism of cytotoxicity and apoptosis induction of the extract at the first hours of incubation and low concentrations were dependent on ROS. However, the toxicity was replaced with NO pathway with time-lapse and higher concentrations. Results also indicated that the extract acts as an NF-κB inhibitor with concentration and time-dependent manners. The present study may suggest the anti-NF-κB activity of PO along with two upstream ROS and NO mechanisms. Furthermore, the extract as ethnobotanical may be used as adjunctive anti-cancer therapy against glioblastoma multiforme.

Aspirin inhibits cancer stem cells properties and growth of glioblastoma multiforme through Rb1 pathway modulation.

Several clinical studies indicated that the daily use of aspirin or acetylsalicylic acid reduces the cancer risk via cyclooxygenases (Cox-1 and Cox-2) inhibition. In addition, aspirin-induced Cox-dependent and -independent antitumor effects have also been described. Here we report, for the first time, that aspirin treatment of human glioblastoma cancer (GBM) stem cells, a small population responsible for tumor progression and recurrence, is associated with reduced cell proliferation and motility. Aspirin did not interfere with cell viability but induced cell-cycle arrest. Exogenous prostaglandin E2 significantly increased cell proliferation but did not abrogate the aspirin-mediated growth inhibition, suggesting a Cox-independent mechanism. These effects appear to be mediated by the increase of p21 waf1 and p27 Kip1 , associated with a reduction of Cyclin D1 and Rb1 protein phosphorylation, and involve the downregulation of key molecules responsible for tumor development, that is, Notch1, Sox2, Stat3, and Survivin. Our results support a possible role of aspirin as adjunctive therapy in the clinical management of GBM patients.