scholarly journals Cytotoxicity Effect of Quinoin, Type 1 Ribosome-Inactivating Protein from Quinoa Seeds, on Glioblastoma Cells

Toxins ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 684
Author(s):  
Rossella Rotondo ◽  
Sara Ragucci ◽  
Salvatore Castaldo ◽  
Maria Antonietta Oliva ◽  
Nicola Landi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Chemotherapeutic Agent ◽  
Primary Cells ◽  
Glioblastoma Cells ◽  
Ribosome Inactivating Protein ◽  
Primary Glioblastoma ◽  
Ribosome Inactivating Proteins ◽  
Primary Cell Lines ◽  
Target Cancer

Ribosome-inactivating proteins (RIPs) are found in several edible plants and are well characterized. Many studies highlight their use in cancer therapy, alone or as immunoconjugates, linked to monoclonal antibodies directed against target cancer cells. In this context, we investigate the cytotoxicity of quinoin, a novel type 1 RIP from quinoa seeds, on human continuous and primary glioblastoma cell lines. The cytotoxic effect of quinoin was assayed on human continuous glioblastoma U87Mg cells. Moreover, considering that common conventional glioblastoma multiforme (GBM) cell lines are genetically different from the tumors from which they derive, the cytotoxicity of quinoin was subsequently tested towards primary cells NULU and ZAR (two cell lines established from patients’ gliomas), also in combination with the chemotherapeutic agent temozolomide (TMZ), currently used in glioblastoma treatment. The present study demonstrated that quinoin (2.5 and 5.0 nM) strongly reduced glioblastoma cells’ growth. The mechanisms responsible for the inhibitory action of quinoin are different in the tested primary cell lines, reproducing the heterogeneous response of glioblastoma cells. Interestingly, primary cells treated with quinoin in combination with TMZ were more sensitive to the treatment. Overall, our data highlight that quinoin could represent a novel tool for glioblastoma therapy and a possible adjuvant for the treatment of the disease in combination with TMZ, alone or as possible immunoconjugates/nanoconstructs.

scholarly journals Assessment of Intratumoral Heterogeneity in Isolated Human Primary High-Grade Glioma: Cluster of Differentiation 133 and Cluster of Differentiation 15 Double Staining of Glioblastoma Subpopulations

2021 ◽  
Vol 9 (A) ◽  
pp. 87-94
Author(s):  
Ahmad Faried ◽  
Wahyu Widowati ◽  
Rizal Rizal ◽  
Hendrikus M. B. Bolly ◽  
Danny Halim ◽  
...  
Keyword(s):  
Brain Tumors ◽  
Cell Line ◽  
Cell Lines ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Double Staining ◽  
Glioblastoma Cells ◽  
Ki 67 ◽  
Primary Glioblastoma ◽  
Cluster Of Differentiation

BACKGROUND: Gliomas are the most common primary brain tumors, representing 50–60% of malignant primary brain tumors. Gliomas are highly heterogeneous with marked inter- and intratumoral diversity. Gliomas heterogeneity is a challenging issue in the development of personalized treatment. The simplest method for studying heterogeneity is using ex vivo cell cultures; in our case, the cell lines were isolated from patient with glioblastomas. AIM: Here, we reported distinct cell subpopulations heterogeneity in glioblastoma cells. METHODS: Human glioblastoma cells isolation is conducted by enzymatic method with combination of collagenase I, hyaluronidase, and trypsin enzyme in proportional amount from patient. Immunostaining was performed to assess glial fibrillary acidic protein (GFAP), Ki-67, isocitrate dehydrogenase-1 (IDH-1) status, and program death ligand-1 (PD-L1) expression. Primary glioblastoma cell line was characterized by flow cytometry (fluorescence-activated cell sorting) analysis based on cluster of differentiation (CD) 133 and CD15 marker expression. U87MG and CGNH-89 cell lines were used as control. Distinct subpopulation analysis was performed by double staining of CD133 and CD15 in isolated primary glioblastoma cell line and its comparative control cells. RESULTS: Our isolated glioblastoma cells morphology was adherent cells which were able to form spheres depending on environment. Immunostaining confirmed GFAP, Ki-67, IDH-1 mutants, and PD-L1 expression. Our isolated glioblastoma cells expressed CD133 and CD15, coexpressed CD133/CD15 in different patterns. The highest subpopulation in primary glioblastoma was CD133+/CD15+. CONCLUSION: Glioblastoma cells can be isolated using enzymatic methods. Isolated glioblastoma cells consist of four different subpopulations distinguished by CD133/CD15 double staining. Intratumoral heterogeneity exists and directly or indirectly depends on their microenvironment.

scholarly journals In Vitro/In Silico Study on the Role of Doubling Time Heterogeneity among Primary Glioblastoma Cell Lines

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
M.-E. Oraiopoulou ◽  
E. Tzamali ◽  
G. Tzedakis ◽  
A. Vakis ◽  
J. Papamatheakis ◽  
...  
Keyword(s):  
Cell Lines ◽  
In Silico ◽  
Intratumoral Heterogeneity ◽  
Patient Specific ◽  
Cell Physiology ◽  
Glioblastoma Cell ◽  
Glioblastoma Cells ◽  
Primary Glioblastoma ◽  
Glioblastoma Cell Lines

The application of accurate cancer predictive algorithms validated with experimental data is a field concerning both basic researchers and clinicians, especially regarding a highly aggressive form of cancer, such as Glioblastoma. In an aim to enhance prediction accuracy in realistic patient-specific environments, accounting for both inter- and intratumoral heterogeneity, we use patient-derived Glioblastoma cells from different patients. We focus on cell proliferation using in vitro experiments to estimate cell doubling times and sizes for established primary Glioblastoma cell lines. A preclinically driven mathematical model parametrization is accomplished by taking into account the experimental measurements. As a control cell line we use the well-studied U87MG cells. Both in vitro and in silico results presented support that the variance between tumor staging can be attributed to the differential proliferative capacity of the different Glioblastoma cells. More specifically, the intratumoral heterogeneity together with the overall proliferation reflected in both the proliferation rate and the mechanical cell contact inhibition can predict the in vitro evolution of different Glioblastoma cell lines growing under the same conditions. Undoubtedly, additional imaging techniques capable of providing spatial information of tumor cell physiology and microenvironment will enhance our understanding regarding Glioblastoma nature and verify and further improve our predictability.

scholarly journals Type 1 ribosome-inactivating proteins are the most abundant proteins in iris (Iris hollandica var. Professor Blaauw) bulbs: characterization and molecular cloning

1997 ◽  
Vol 324 (3) ◽  
pp. 963-970 ◽  
Author(s):  
Els J. M. VAN DAMME ◽  
Annick BARRE ◽  
Luigi BARBIERI ◽  
Paola VALBONESI ◽  
Pierre ROUGE ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Nitrogen Storage ◽  
Related Protein ◽  
Ribosome Inactivating Protein ◽  
Storage Organ ◽  
Abundant Protein ◽  
Ribosome Inactivating Proteins ◽  
Sequence Identity ◽  
High Degree

The most abundant protein of Iris bulbs has been identified as a type 1 ribosome-inactivating protein (RIP). Analysis of the purified proteins and molecular cloning of the corresponding cDNAs demonstrated that this type 1 RIP is a mixture of three isoforms that exhibit a high degree of sequence identity and have similar, though not identical, ribosome-inactivating and polynucleotide:adenosine glycosidase activities. The accumulation of large quantities of type 1 RIP in a vegetative storage organ suggests that this presumed defence-related protein also plays a role in the nitrogen-storage metabolism of the bulb.

Fructose metabolism in hepatocytes: primary cells or cell lines?

2007 ◽  
Vol 45 (01) ◽  
Author(s):  
T Speicher ◽  
G Künstle ◽  
A Wendel
Keyword(s):  
Cell Lines ◽  
Primary Cells ◽  
Fructose Metabolism

Glioblastoma invasion and NMDA receptors: A novel prospect

2019 ◽  
Vol 106 (3) ◽  
pp. 250-260 ◽  
Author(s):  
DN Nandakumar ◽  
P Ramaswamy ◽  
C Prasad ◽  
D Srinivas ◽  
K Goswami
Keyword(s):  
Glutamate Receptors ◽  
Cell Lines ◽  
Intracellular Signaling ◽  
Transcript Level ◽  
Glioblastoma Cells ◽  
Invasion And Migration ◽  
Matrigel Invasion ◽  
Nmdar Activation ◽  
And Migration

Purpose Glioblastoma cells create glutamate-rich tumor microenvironment, which initiates activation of ion channels and modulates downstream intracellular signaling. N-methyl-D-aspartate receptors (NMDARs; a type of glutamate receptors) have a high affinity for glutamate. The role of NMDAR activation on invasion of glioblastoma cells and the crosstalk with α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs) is yet to be explored. Main methods LN18, U251MG, and patient-derived glioblastoma cells were stimulated with NMDA to activate NMDAR glutamate receptors. The role of NMDAR activation on invasion and migration and its crosstalk with AMPAR were evaluated. Invasion and migration of glioblastoma cells were investigated by in vitro trans-well Matrigel invasion and trans-well migration assays, respectively. Expression of NMDARs and AMPARs at transcript level was evaluated by quantitative real-time polymerase chain reaction. Results We determined that NMDA stimulation leads to enhanced invasion in LN18, U251MG, and patient-derived glioblastoma cells, whereas inhibition of NMDAR using MK-801, a non-competitive antagonist of the NMDAR, significantly decreased the invasive capacity. Concordant with these findings, migration was significantly augmented by NMDAR in both cell lines. Furthermore, NMDA stimulation upregulated the expression of GluN2 and GluA1 subunits at the transcript level. Conclusions This study demonstrated the previously unexplored role of NMDAR in invasion of glioblastoma cells. Furthermore, the expression of the GluN2 subunit of NMDAR and the differential overexpression of the GluA1 subunit of AMPAR in both cell lines provide a plausible rationale of crosstalk between these calcium-permeable subunits in the glutamate-rich microenvironment of glioblastoma.

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