scholarly journals Albumin–Methotrexate Prodrug Analogues That Undergo Intracellular Reactivation Following Entrance into Cancerous Glioma Cells

Pharmaceutics ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 71
Author(s):  
Itzik Cooper ◽  
Michal Schnaider-Beeri ◽  
Mati Fridkin ◽  
Yoram Shechter
Keyword(s):  
Dihydrofolate Reductase ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Peptide Bond ◽  
Glioma Cell Line ◽  
Native Structure ◽  
Free System ◽  
A Cell ◽  
Covalently Linked ◽  
Single Peptide

A family of monomodified bovine serum albumin (BSA) linked to methotrexate (MTX) through a variety of spacers was prepared. All analogues were found to be prodrugs having low MTX-inhibitory potencies toward dihydrofolate reductase in a cell-free system. The optimal conjugates regenerated their antiproliferative efficacies following entrance into cancerous glioma cell lines and were significantly superior to MTX in an insensitive glioma cell line. A BSA–MTX conjugate linked through a simple ethylene chain spacer, containing a single peptide bond located 8.7 Å distal to the protein back bone, and apart from the covalently linked MTX by about 12 Å, was most effective. The inclusion of an additional disulfide bond in the spacer neither enhanced nor reduced the killing potency of this analogue. Disrupting the native structure of the carrier protein in the conjugates significantly reduced their antiproliferative activity. In conclusion, we have engineered BSA–MTX prodrug analogues which undergo intracellular reactivation and facilitate antiproliferative activities following their entrance into glioma cells.

Enhancement of radiosensitivity of wild-type p53 human glioma cells by adenovirus-mediated delivery of the p53 gene

1998 ◽  
Vol 89 (1) ◽  
pp. 125-132 ◽  
Author(s):  
Frederick F. Lang ◽  
W. K. Alfred Yung ◽  
Uma Raju ◽  
Floralyn Libunao ◽  
Nicholas H. A. Terry ◽  
...  
Keyword(s):  
Cell Line ◽  
Glioma Cell ◽  
Radiation Treatment ◽  
P53 Protein ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Wild Type ◽  
Content Type ◽  
Infected Cells ◽  
Fine Print

Object. The authors sought to determine whether combining p53 gene transfer with radiation therapy would enhance the therapeutic killing of p53 wild-type glioma cells. It has been shown in several reports that adenovirus-mediated delivery of the p53 gene into p53 mutant gliomas results in dramatic apoptosis, but has little effect on gliomas containing wild-type p53 alleles. Therefore, p53 gene therapy alone may not be a clinically effective treatment for gliomas because most gliomas are composed of both p53 mutant and wild-type cell populations. One potential approach to overcome this problem is to exploit the role p53 plays as an important determinant in the cellular response to ionizing radiation. Methods. In vitro experiments were performed using the glioma cell line U87MG, which contains wild-type p53. Comparisons were made to the glioma cell line U251MG, which contains a mutant p53 allele. Monolayer cultures were infected with an adenovirus containing wild-type p53 (Ad5CMV-p53), a control vector (dl312), or Dulbecco's modified Eagle's medium (DMEM). Two days later, cultures were irradiated and colony-forming efficiency was determined. Transfection with p53 had only a minor effect on the plating efficiency of nonirradiated U87MG cells, reducing the plating efficiency from 0.23 ± 0.01 in DMEM to 0.22 ± 0.04 after addition of Ad5CMV-p53. However, p53 transfection significantly enhanced the radiosensitivity of these cells. The dose enhancement factor at a surviving fraction of 0.10 was 1.5, and the surviving fraction at 2 Gy was reduced from 0.61 in untransfected controls to 0.38 in p53-transfected cells. Transfection of the viral vector control (dl312) had no effect on U87MG radiosensitivity. In comparison, transfection of Ad5CMV-p53 into the p53 mutant cell line U251MG resulted in a significant decrease in the surviving fraction of these cells compared with controls, and no radiosensitization was detected. To determine whether Ad5CMV-p53—mediated radiosensitization of U87MG cells involved an increase in the propensity of these cells to undergo apoptosis, flow cytometric analysis of terminal deoxynucleotidyl transferase-mediated biotinylated-deoxyuridinetriphosphate nick-end labeling—stained cells was performed. Whereas the amount of radiation-induced apoptosis in uninfected and dl312-infected control cells was relatively small (2.1 ± 0.05% and 3.7 ± 0.5%, respectively), the combination of Ad5CMV-p53 infection and radiation treatment significantly increased the apoptotic frequency (18.6 ± 1.4%). To determine whether infection with Ad5CMV-p53 resulted in increased expression of functional exogenous p53 protein, Western blot analysis of p53 was performed on U87MG cells that were exposed to 9 Gy of radiation 2 days after exposure to Ad5CMV-p53, dl312, or DMEM. Infection with Ad5CMV-p53 alone increased p53 levels compared with DMEM- or dl312-treated cells. Irradiation of Ad5CMV-p53—infected cells resulted in a further increase in p53 that reached a maximum at 2 hours postirradiation. To determine whether exogenous p53 provided by Ad5CMV-p53 had transactivating activity, U87MG cells were treated as described earlier and p21 messenger RNA levels were determined. Infection of U87MG cells with Ad5CMV-p53 only resulted in an increase in p21 compared with DMEM- and dl312-treated cells. Irradiation of Ad5CMV-p53—infected cells resulted in an additional time-dependent increase in p21 expression. Conclusions. These data indicate that adenovirus-mediated delivery of p53 may enhance the radioresponse of brain tumor cells containing wild-type p53 and that this radiosensitization may involve converting from a clonogenic to the more sensitive apoptotic form of cell death. Although the mechanism underlying this enhanced apoptotic susceptibility is unknown, the Ad5CMV-p53—infected cells have a higher level of p53 protein, which increases further after irradiation, and this exogenous p53 is transcriptionally active. Thus, it is possible that the combination of Ad5CMV-p53 infection and radiation treatment increases p53 protein to a level that is sufficient to overcome at least partially the block in apoptosis existing in U87MG cells.

scholarly journals Single-Cell Mechanophenotyping in Microfluidics to Evaluate Behavior of U87 Glioma Cells

Micromachines ◽  
2020 ◽  
Vol 11 (9) ◽  
pp. 845
Author(s):  
Esra Sengul ◽  
Meltem Elitas
Keyword(s):  
Single Cell ◽  
Glioma Cell ◽  
Single Cells ◽  
Glioma Cells ◽  
Population Level ◽  
Glioma Cell Line ◽  
Scientific Discipline ◽  
Regular Growth ◽  
Mesenchymal Transition ◽  
Deformation Properties

Integration of microfabricated, single-cell resolution and traditional, population-level biological assays will be the future of modern techniques in biology that will enroll in the evolution of biology into a precision scientific discipline. In this study, we developed a microfabricated cell culture platform to investigate the indirect influence of macrophages on glioma cell behavior. We quantified proliferation, morphology, motility, migration, and deformation properties of glioma cells at single-cell level and compared these results with population-level data. Our results showed that glioma cells obtained slightly slower proliferation, higher motility, and extremely significant deformation capability when cultured with 50% regular growth medium and 50% macrophage-depleted medium. When the expression levels of E-cadherin and Vimentin proteins were measured, it was verified that observed mechanophenotypic alterations in glioma cells were not due to epithelium to mesenchymal transition. Our results were consistent with previously reported enormous heterogeneity of U87 glioma cell line. Herein, for the first time, we quantified the change of deformation indexes of U87 glioma cells using microfluidic devices for single-cells analysis.

Effect of Geldanamycin on the Glioma Cell Invasion Induced by Hepatocyte Growth Factor

2014 ◽  
Vol 1033-1034 ◽  
pp. 224-228
Author(s):  
Yan Xia Sun ◽  
Guang Yu Zhou ◽  
Li Li ◽  
Xue Mei Han
Keyword(s):  
Growth Factor ◽  
Hepatocyte Growth Factor ◽  
Cell Invasion ◽  
Culture System ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Invasive Growth ◽  
Transwell Culture ◽  
Hepatocyte Growth

To investigate the effect of geldanamycin (GDM) on the invasion ability of glioma cell induced by hepatocyte growth factor (HGF). Malignant glioma cell line U251-MG and U87-MG were cultured’and the capability of cell invasion was detected using a Transwell culture system. HGF significantly promoted the invasion ability of both U251-MG and U87-MG cells as compared with the normal control (NC) (P < 0.05). Forty eight hours after GDM treatment, the invasive growth of glioma cells was significantly decreased as compared with either NC or HGF group (P < 0.05). When cells were exposed to GDM plus HGF for 48 h, the cell invasion capability was greatly reduced as compared with either NC or HGF group (P < 0.05). The number of invaded cells in GDM plus HGF group was similar to that of GDM group.GDM can inhibit the invasion ability of glioma cells induced by HGF.

scholarly journals HIV-1 Envelope Protein gp120 Promotes Proliferation and the Activation of Glycolysis in Glioma Cell

Cancers ◽  
2018 ◽  
Vol 10 (9) ◽  
pp. 301 ◽  
Author(s):  
Gabriel Valentín-Guillama ◽  
Sheila López ◽  
Yuriy Kucheryavykh ◽  
Nataliya Chorna ◽  
Jose Pérez ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Envelope Glycoprotein ◽  
Glioma Cell ◽  
Lipid Synthesis ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Atp Production ◽  
Immunodeficiency Virus ◽  
Glycoprotein Gp120 ◽  
Hiv Envelope

Patients infected with human immunodeficiency virus (HIV) are more prone to developing cancers, including glioblastomas (GBMs). The median survival for HIV positive GBM patients is significantly shorter than for those who are uninfected, despite the fact that they receive the same treatments. The nature of the GBM–HIV association remains poorly understood. In this study, we analyzed the effect of the HIV envelope glycoprotein gp120 on GBM cell proliferation. Specifically, we performed cell cycle, western blot, protein synthesis and metabolomics analysis as well as ATP production and oxygen consumption assays to evaluate proliferation and metabolic pathways in primary human glioma cell line, U87, A172 cells and in the HIVgp120tg/GL261 mouse model. Glioma cells treated with gp120 (100 ng/mL for 7–10 days) showed higher proliferation rates and upregulation in the expression of enolase 2, hexokinase and glyceraldehyde-3-phosphate dehydrogenase when compared to untreated cells. Furthermore, we detected an increase in the activity of pyruvate kinase and a higher glycolytic index in gp120 treated cells. Gp120 treated GBM cells also showed heightened lipid and protein synthesis. Overall, we demonstrate that in glioma cells, the HIV envelope glycoprotein promotes proliferation and activation of glycolysis resulting in increased protein and lipid synthesis.

scholarly journals Effects of miR-99a on the migration and proliferation of glioma cells

2018 ◽  
Vol 26 (2) ◽  
pp. 145-152
Author(s):  
Yifan Xu ◽  
Tianyu Lu ◽  
Wu Xu ◽  
Yuxiang Dai ◽  
Weibang Liang ◽  
...  
Keyword(s):  
Cell Line ◽  
Mtt Assay ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Scratch Assay ◽  
Knock Down ◽  
And Migration ◽  
Proliferation And Migration

Abstract Background To evaluate the effects of miR-99a on the migration and proliferation of glioma cells. Materials and Methods: Glioma cell line LN229 with stable up-regulation of miR-99a was constructed by transfection of hsa-miR-99a mimics, and cells with stable miR-99a knock-down were established by transfection of hsa-miR-99a inhibitor. The proliferation capacities of two groups were detected by the MTT assay, and their migration capacities were detected by the scratch assay. Results: LN229 cells with stable up-regulation and knock-down of miR-99a were successfully constructed. Up-regulating miR-99a inhibited the proliferation and migration of glioma cells, but knocking down this gene promoted their proliferation and migration. Conclusion: MiR-99a significantly affected the proliferation and migration of glioma cells, as a potentially eligible target for glioma therapy.

scholarly journals The primary culture of malignant glioma cells as a model for the study of anti-tumor activity of substances

2020 ◽  
Vol 17 (2) ◽  
pp. 196-203
Author(s):  
I. M. Shuba ◽  
V. V. Lylo ◽  
I. S. Karpova ◽  
O. Y. Glavatskyi ◽  
O. I. Kornelyuk
Keyword(s):  
Cell Culture ◽  
Malignant Glioma ◽  
Primary Culture ◽  
Cell Line ◽  
Glioma Cell ◽  
Primary Cell Culture ◽  
Malignant Gliomas ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Malignant Glioma Cells

Aim. The aim of our work was to optimize the scheme of obtaining primary cell culture of malignant gliomas, which can be a model for a personalized approach in the selection of chemotherapeutic exposure tactics. Methods. The standard glioma cell line U-251MG and cells obtained as a result of mechanical disaggregation of Gr III–IV tumor fragments to single isolated cells were used. Results. A comparative analysis of the results of cultivation of the standard glioma cell line U-251MG and the primary cell culture of malignant gliomas. An optimized scheme for obtaining primary cultures of human malignant glioma cells isolated from glial tumor fragments obtained during surgery is proposed. Conclusions. Today, more and more preferred methods of individual determination of chemosensitivity over the appointment of standard chemotherapy regimens and it is the primary culture of tumor cells, from our point of view, can be used to test the response to the effect of chemotherapy.Keywords: malignant glioma cells, primary culture, standard cell line.

COMPARATIVE ANALYSIS OF THE BINDING EFFICIENCY OF TUMOR- TARGETING PHAGE PARTICLES INTO CANCER CELLS AND HEALTH BRAIN CELLS

2020 ◽  
pp. 23-24
Author(s):  
A. Voitova ◽  
M. Dmitrieva ◽  
V. Richter ◽  
E. Kuligina
Keyword(s):  
Flow Cytometry ◽  
Glioma Cell ◽  
Tumor Targeting ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Enzyme Linked Immunosorbent Assay ◽  
Brain Cells ◽  
Human Glioma ◽  
Human Glioma Cells

The binding efficiency of tumor-targeting phage particles, obtained by phage display, into human glioma cell line U-87 MG and health brain cells was evaluated by flow cytometry and enzyme-linked immunosorbent assay (ELISA). Based on the obtained data, tumor-targeting phage particles that provide the most efficient binding to human glioma cells U-87 MG in vitro are selected for further studies.

scholarly journals Differentiation of Tumorigenic C6 Glioma Cells Induced by Enhanced IL-6 Signaling

Medicina ◽  
2020 ◽  
Vol 56 (11) ◽  
pp. 625
Author(s):  
Inn-Ray Chu ◽  
Rong-Long Pan ◽  
Chung-Shi Yang
Keyword(s):  
Glioma Cell ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
C6 Glioma ◽  
C6 Glioma Cells ◽  
Differentiation Therapy ◽  
C6 Glioma Cell ◽  
Tnf Α ◽  
Total Cell Population ◽  
Factor Α

Background and objectives: Cancer stem cells (CSCs) are obstacles to cancer therapy due to their therapeutic resistance, ability to initiate neoplasia, and roles in tumor relapse and metastasis. Efforts have been made to cure CSCs, such as the use of differentiation therapy, which induces cancer stem-like cells to undergo differentiation and decrease their tumorigenicity. Interleukin 6 (IL-6) upregulates the expression of glial fibrillary acidic protein (GFAP) in C6 glioma cells, indicating that it is able to induce the differentiation of these cells. The C6 glioma cell line forms a high percentage of cancer stem-like cells, leading us to speculate whether IL-6 signaling could modulate the differentiation of tumorigenic C6 glioma cells. However, we observed that IL-6 alone could not efficiently induce the differentiation of these cells. Therefore, different IL-6 signaling elicitors, including IL-6 alone, a combination of IL-6 and soluble IL-6 receptor (IL-6/sIL-6R), and tumor necrosis factor-α (TNF-α) plus IL-6/sIL-6R (TNF-α/IL-6/sIL-6R), were evaluated for their potential use in differentiation therapy. Materials and Methods: The potential of IL-6 signaling elicitors in differentiation therapy were examined by assessing changes in biomarker levels, the rate of cell proliferation, and tumorigenicity, respectively. Results: Enhanced IL-6 signaling could effectively induce C6 glioma cell differentiation, as determined by observed variations in the expression of differentiation, cell cycle, and stem cell biomarkers. Additionally, the total cell population and the tumorigenicity of glioma cells were all considerably reduced after TNF-α/IL-6/sIL-6R treatment. Conclusions: Our findings provide evidence that enhanced IL-6 signaling can efficiently promote tumorigenic C6 glioma cells to undergo differentiation.

Effect of TWIST1 Gene on the Proliferation and Apoptosis of Human Glioma Cell Line TJ861 by Regulating Mammalian Target of Rapamycin Signaling Pathway

2021 ◽  
Vol 11 (4) ◽  
pp. 580-585
Author(s):  
Fei Chen ◽  
Jiajia Hua ◽  
HongWei Shen ◽  
HongLiang Wang
Keyword(s):  
Protein Expression ◽  
Cell Line ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Glioma Cell Line ◽  
Control Group ◽  
Human Glioma ◽  
Human Glioma Cell Line ◽  
Human Glioma Cell ◽  
Mtor Protein

To observe TWIST1 gene expression in human glioma and study the effect human glioma cell line TJ861 on the proliferation and apoptosis, and further explore its potential mechanism to provide some reference for the targeted treatment of glioma in the future. Detection of cancer tissue (Carcinoma tissue) in 55 patients with glioma by RT-PCR and Expression level of TWIST1 in normal and paracancerous tissues (Adjacent tissue), the human glioma cell line TJ861 was further divided into, Nonsense sequence group, (si-NS group), TWIST1 Inhibition group (si-TWIST1 group) and control group. The glioma cells of si-NS group and si-TWIST1 group were transfected with nonsense sequence and TWIST1 siRNA respectively by liposome transfection technology. Use CCK8 assay to test the cell proliferation ability of each group at 0, 12, 24, 36, 48 and 72 hours; 48 hours after siRNA transfection, The ability of DNA replication in each group was detected by EdU staining; Apoptosis related protein expression, in each group, was analyzed by Western blot; TUNEL staining was used to test the apoptosis rate of each group; In the end, We studied TWIST1 effect knocking down on mTOR protein expression in human glioma cells and mTOR protein expression in cancer and adjacent tissues. TWIST1 expression in glioma cells was higher, compared with normal tissues (P <0.05); After transfection of TWIST1 siRNA into human glioma cell line TJ861 in vitro, CCK8 showed glioma cells proliferation ability in si-TWIST1 group at 12, 24, 36, 48 and 72 hours was lower, compared with the control group (P <0.05); After siRNA transfection at 48 hours, the DNA replication ability of glioma cells decreased significantly (P <0.05) with EdU staining; The inhibition of TWIST1 increased Bax expression in glioma cells, and inhibited Bcl-2 expression (P < 0.05) with Western blot; TUNEL staining further confirmed that the apoptosis level of glioma cells in the si-TWIST1 group was higher, compared with the control group (P <0.05). Finally, we found that mTOR protein expression in glioma was higher, compared with adjacent tissues. in vitro experiments showed that mTOR expression in glioma cells was decreased after the inhibition of TWIST1 (P <0.05). TWIST1 expression level in glioma was increased. The inhibition of TWIST1 inhibits the proliferation of glioma by blocking the mTOR signal pathway, and promote the apoptosis of glioma.

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