Elucidating the Innate Immunological Effects of Mild Magnetic Hyperthermia on U87 Human Glioblastoma Cells: An In Vitro Study

Cancer immunotherapies have been approved as standard second-line or in some cases even as first-line treatment for a wide range of cancers. However, immunotherapy has not shown clinically relevant success in glioblastoma (GBM). This is principally due to the brain’s “immune-privileged” status and the peculiar tumor microenvironment (TME) of GBM characterized by a lack of tumor-infiltrating lymphocytes and the establishment of immunosuppressive mechanisms.
Herein, we explore a local mild thermal treatment, generated via cubic-shaped iron oxide magnetic nanoparticles (size ~17 nm) when exposed to an external alternating magnetic field (AMF), to induce immunogenic cell death (ICD) in U87 glioblastoma cells. In accordance with what has been observed with other tumor types, we found that mild magnetic hyperthermia (MHT) modulates the immunological profile of U87 glioblastoma cells by inducing stress-associated signals leading to enhanced phagocytosis and killing of U87 cells by macrophages.
At the same time, we demonstrated that mild magnetic hyperthermia on U87 cells has a modulatory effect on the expression of inhibitory and activating NK cell ligands. Interestingly, this alteration in the expression of NK ligands in U87 cells upon MHT treatment increased their susceptibility to NK cell killing and enhanced NK cell functionality.
The overall findings demonstrate that mild MHT stimulates ICD and sensitizes GBM cells to NK-mediated killing by inducing the upregulation of specific stress ligands, providing a novel immunotherapeutic approach for GBM treatment, with potential to synergize with existing NK cell-based therapies thus improving their therapeutic outcomes.

Lectin isolated from Abelmoschus esculentus induces caspase mediated apoptosis in Human U87 glioblastoma cell lines and modulates the expression of circadian clock genes

Lectins are a cluster of proteins which are capable of recognizing and binding to glycoconjugates and are extensively found in plants, animals, fungi and bacteria.
Plant-derived lectins have been gaining importance over the years due to their innumerable biological activities and also have the added possibility of being compatible to the human system while simultaneously exhibiting properties like antimicrobial and antitumor activities.
Abelmoschus esculentus (AE) commonly known as okra is a vegetable with medicinal properties. AE extracts are used to treat disorders such as constipation, microbial infection, urine retention, hypoglycemia and inflammation in humans.
Previous studies showed that lectin isolated from AE exhibited anti inflammatory, anti nociceptive, anticancer, antioxidant and hemagglutinating activities.
However, the antitumor effect of the lectin derived from this plant against neural cancer cells still remains unexplored. Glioblastoma is a malignant tumor of the nervous system.
Treatment options for patients afflicted by glioblastoma is limited to surgical resection, preceded by radiation therapy and followed by chemotherapy.
Hence it would be of interest to identify novel bio molecules with ability to selectively target glioblastoma with minimum side effects. In this aspect, lectins from vegetables that are commonly used as food products could offer a promising lead as anticancer molecules.
The present study proves the anti-proliferative effect of lectin isolated from AE on human U87 glioma cells. MTT assay showed significant concentration dependent cytotoxic activity and the IC50 value was calculated as 21μg/ml. Further, annexin V/FITC staining by FACS, the expression of caspase 3 and 7 and the circadian genes clock and Bmal1 using RT-PCR and the generation of intracellular ROS, cell cycle analysis by FACS revealed the ability of AEL to induce effective apoptosis.

Probing mechanobiological role of filamin A in migration and invasion of human U87 glioblastoma cells using submicron soft pillars

Filamin A (FLNa) belongs to an actin-binding protein family in binding and cross-linking actin filaments into a three-dimensional structure. However, little attention has bee
n given to its mechanobiological role in cancer cells. Here, we quantitatively investigated the role of FLNa by analyzing the following parameters in negative control (NC) and FLNa-knockdown (KD) U87 glioma cells using submicron pillars (900 nm diameter and 2 μm height): traction force (TF), rigidity sensing ability, cell aspect ratio, migration speed, and invasiveness.
During the initial phase of cell adhesion (< 1 h), FLNa-KD cells polarized more slowly than did NC cells, which can be explained by the loss of rigidity sensing in FLNa-KD cells.
The higher motility of FLNa-KD cells relative to NC cells can be explained by the high TF exerted by FLNa-KD cells when compared to NC cells, while the higher invasiveness of FLNa-KD cells relative to NC cells can be explained by a greater number of filopodia in FLNa-KD cells than in NC cells. Our results suggest that FLNa plays important roles in suppressing motility and invasiveness of U87 cells.

WDR81 Gene Silencing Can Reduce Exosome Levels in Human U87-MG Glioblastoma Cells

Glioblastoma is a very invasive and prevalent brain tumor that affects 15 in 100,000 persons over the age of 70 years. Studies have shown that the expression of the WD repeat domain 81 (WDR81) gene, which is effective in vesicular transport and inhibition of autophagy, is increased in glioblastoma.
The decreased autophagy was found to be related to the increased production of exosomes, which is a major factor in the pathogenesis of glioblastoma. The PI-3kinase complex is a pre-autophagic complex that is highly active in the absence of WDR81.
The WDR81 gene, as a negative regulator of PI3K activity, prevents autophagy and increases exosome secretion by preventing the formation of the class III PI3K complex. Therefore, targeted reduction of exosomes can be considered an effective strategy for reducing the pathogenesis of glioblastoma.
This study aimed to assess the effect of WDR81 gene silencing with siRNA on exosome levels in a U87-MG cell line. Culturing of U87-MG cells was carried out in Dulbecco’s modified Eagle medium (DMEM) containing 5% FBS and 1% penicillin/streptomycin. Thereafter, silencing of WDR81 was performed using WDR81 siRNA, whose gene expression level was determined via real-time qRT-PCR. Cell viability was evaluated using the MTT assay.
The exosomes were extracted from a cell culture using the Exocib kit. The size accuracy of the exosomes was confirmed by dynamic light scattering (DLS). Finally, the protein content and RNA of the exosomes were assessed. WDR81 gene expression of siRNA-transfected cells was decreased to 82% after 24 h compared to the non-transfected control cells.
The analysis of the exosomes showed that the concentration of exosomes and their RNA and protein content in the siRNA-transfected cells decreased significantly compared to the non-transfected control cells. No considerable difference was observed in cell viability after transfection with either WDR81-specific siRNAs or scrambled control siRNAs.
Our findings showed that silencing the WDR81 gene could reduce the level of exosomes in human U87-MG glioblastoma cells. Therefore, the reduced exosome content may be suggested as a new gene therapy strategy for targeted therapy of glioblastoma by increasing autophagy via activation of PI3KIII. However, more studies are needed in this regard.

Enantioselective effect of cysteine functionalized mesoporous silica nanoparticles in U87 MG and GM08680 human cells and Staphylococcus aureus bacteria

 

Chirality is a fundamental phenomenon in biological systems, since most of the biomolecules and biological components and species are chiral and therefore recognize and respond differently depending on the enantiomer present. With increasing research into the use of nanomaterials for biomedical purposes, it is essential to understand the role that chirality of nanoparticles plays at the cellular level.

Human U87 Cell Lysate

from BosterBio
LYSATE0041 | 200ug: 180.00 EUR

ExoStd? U87 MG Fluorescent Exosome Standard

from Biovision
M1084-100 | : 907.20 EUR

ExoStd? Lyophilized Exosome Standard (30 µg, U87 MG, 2 vials)

from Biovision
M1054-2 | : 724.80 EUR

ExoStd? Lyophilized Exosome Standard (30 µg, U87 MG, 4 vials)

from Biovision
M1054-4 | : 1096.80 EUR

ExoStd? Lyophilized Exosome Standard (30 µg, U87 MG, 6 vials)

from Biovision
M1054-6 | : 1567.20 EUR

ExoStd? Lyophilized Exosome Standard (100 µg, U87 MG, 2 vials)

from Biovision
M1055-2 | : 861.60 EUR

ExoStd? Lyophilized Exosome Standard (100 µg, U87 MG, 4 vials)

from Biovision
M1055-4 | : 1370.40 EUR

ExoStd? Lyophilized Exosome Standard (100 µg, U87 MG, 6 vials)

from Biovision
M1055-6 | : 1939.20 EUR

Human CellExp? Human CD40/ TNFRSF5, Human recombinant

from Biovision
9230-10 | : 333.60 EUR

Human CellExp? Human CD40/ TNFRSF5, Human recombinant

from Biovision
9230-50 | : 868.80 EUR

Human CellExp? Renin, Human Recombinant

from Biovision
6300-10 | : 196.80 EUR

Human CellExp? Renin, Human Recombinant

from Biovision
6300-100 | : 940.80 EUR

Human CellExp? Renin, Human Recombinant

from Biovision
6300-50 | : 601.20 EUR

Human CellExp? EPO, Human Recombinant

from Biovision
6447-10 | : 646.80 EUR

Human CellExp? EPO, Human Recombinant

from Biovision
6447-50 | : 2017.20 EUR

Human CellExp? HGF, Human Recombinant

from Biovision
6456-10 | : 464.40 EUR

Human CellExp? HGF, Human Recombinant

from Biovision
6456-50 | : 1677.60 EUR

Human CellExp? HGH, Human Recombinant

from Biovision
6457-10 | : 405.60 EUR

Human CellExp? HGH, Human Recombinant

from Biovision
6457-50 | : 1455.60 EUR

Human CellExp? Noggin, Human Recombinant

from Biovision
6474-10 | : 360.00 EUR

Human CellExp? Noggin, Human Recombinant

from Biovision
6474-50 | : 1286.40 EUR

Human CellExp? SCF, Human Recombinant

from Biovision
6478-10 | : 411.60 EUR
Here, the chiral cysteine functionalization of mesoporous silica nanoparticles has been shown to broadly affect its interaction with U87 MG human glioblastoma cell, healthy human fibroblast (GM08680) and methicillin-resistant S. aureus bacteria. We believe that this research is important to further advancement of nano-biotechnology.

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