scholarly journals P2X7 receptor activity landscape in rat and human glioma cell lines

2020 ◽  
Author(s):  
Damian Matyśniak ◽  
Natalia Nowak ◽  
Vira Chumak ◽  
Paweł Pomorski
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Human Cell ◽  
P2x7 Receptor ◽  
Pore Formation ◽  
Glioma Cell ◽  
Purinergic Receptor ◽  
Receptor Expression ◽  
Calcium Signal ◽  
Glioma Cell Lines

P2X7 is a commonly expressed purinergic receptor, which functions as a cation-permeable channel in the plasma membrane. In certain circumstances, the receptor may also form a large transmembrane pore what results in cell death. P2X7 receptors control numerous physiological and pathological cellular processes and their overexpression is often associated with cancer progression. As nucleotides are important signaling molecules in the central nervous system, P2X7 plays also an important but ambiguous role in glioma biology with contrary observations originating from different glioma models. Therefore, the aim of our research was to investigate P2X7 receptor expression and functions in three human (U-87 MG, U-138 MG, U-251 MG) and one rat (C6) glioma cell lines. Although the receptor mRNA and protein were present in all the studied cells, we found profound differences in their level. We also encountered a problem with one human cell lines authenticity (U-87 MG) and excluded it from most of the experiments. Interestingly, there was no clear dependency between P2X7 receptor level, calcium signal and pore formation ability in the studied glioma lines. In U-138 human cell line, the receptor seemed to be inactive, while in U-251 human and C6 rat cell line its activation resulted in calcium influx and large pore formation. However, the viability of studied cells upon the administration of specific P2X7 agonist – BzATP – was not affected for U-138 and U-251, whereas for C6 cells a stimulatory effect was observed. Our results stress the variability of P2X7 signaling in glioma models and the need for future research which would take into account the complicated landscape of the receptor signaling in the brain.

scholarly journals Temozolomide sensitivity of malignant glioma cell lines – a systematic review assessing consistencies between in vitro studies

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Michael T. C. Poon ◽  
Morgan Bruce ◽  
Joanne E. Simpson ◽  
Cathal J. Hannan ◽  
Paul M. Brennan
Keyword(s):  
Cell Viability ◽  
Malignant Glioma ◽  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Glioma Cell Line ◽  
In Vitro Studies ◽  
Experimental Conditions ◽  
Glioma Cell Lines

Abstract Background Malignant glioma cell line models are integral to pre-clinical testing of novel potential therapies. Accurate prediction of likely efficacy in the clinic requires that these models are reliable and consistent. We assessed this by examining the reporting of experimental conditions and sensitivity to temozolomide in glioma cells lines. Methods We searched Medline and Embase (Jan 1994-Jan 2021) for studies evaluating the effect of temozolomide monotherapy on cell viability of at least one malignant glioma cell line. Key data items included type of cell lines, temozolomide exposure duration in hours (hr), and cell viability measure (IC50). Results We included 212 studies from 2789 non-duplicate records that reported 248 distinct cell lines. The commonest cell line was U87 (60.4%). Only 10.4% studies used a patient-derived cell line. The proportion of studies not reporting each experimental condition ranged from 8.0–27.4%, including base medium (8.0%), serum supplementation (9.9%) and number of replicates (27.4%). In studies reporting IC50, the median value for U87 at 24 h, 48 h and 72 h was 123.9 μM (IQR 75.3–277.7 μM), 223.1 μM (IQR 92.0–590.1 μM) and 230.0 μM (IQR 34.1–650.0 μM), respectively. The median IC50 at 72 h for patient-derived cell lines was 220 μM (IQR 81.1–800.0 μM). Conclusion Temozolomide sensitivity reported in comparable studies was not consistent between or within malignant glioma cell lines. Drug discovery science performed on these models cannot reliably inform clinical translation. A consensus model of reporting can maximise reproducibility and consistency among in vitro studies.

scholarly journals A Novel Gene RNF138 Expressed in Human Gliomas and Its Function in the Glioma Cell Line U251

2012 ◽  
Vol 35 (3) ◽  
pp. 167-178 ◽  
Author(s):  
You-xin Zhou ◽  
San-song Chen ◽  
Ting-feng Wu ◽  
Da-dong Ding ◽  
Xiong-hui Chen ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Cell Line ◽  
Brain Tissue ◽  
Cell Lines ◽  
Cell Apoptosis ◽  
Glioma Cell ◽  
Glioma Cell Line ◽  
Glioma Tissue ◽  
Glioma Cell Lines

Background: The gliomas represent the most common primary malignant brain tumors; however, little is known about the molecular pathogenesis of these tumors. Recent research reveals that the oncogenesis and development of gliomas have a close relation to the overexpression of several oncogenes and the inactivation of tumor suppressor genes. Whether the RING finger protein, RNF138, a newly discovered protein, plays a role in glioma oncogenesis is unknown. The present study investigates the expression levels of RNF138 mRNA in glioma samples and noncancerous brain samples and its function in the human glioma cell line U251.Methods: RT-PCR was used to ascertain the expression of RNF138 mRNA in the glioma cell lines U251, SHG44, U87, A172, and U373. The RNF138 mRNA expression levels of 35 pathological confirmed glioma samples (Grade I – 4 cases, Grade II – 13 cases, Grade III – 11 cases, and Grade IV – 7 cases) and five noncancerous brain tissue samples were analyzed by real-time quantitative PCR. By RNA interference (RNAi) with the lentivirus vector system, the expression of RNF138 was inhibited in the human astrocytomas-glioblastoma multiforme cell line U251. The effects of RNF138-knockdown on cell proliferation were assessed by Cellomics, and cell cycle and cell apoptosis were assessed by FACS.Results: The RNF138 mRNA is expressed in the five glioma cell lines, and its expression level is significantly higher in glioma tissue than in noncancerous brain tissue. By down-regulation of RNF138 expression, U251 cell proliferation was inhibited and cell apoptosis increased. At the same time, S stage cells lessened and G2 stage cells increased.Conclusion: The RNF138 gene is highly expressed in glioma tissue and glioma cell lines. It plays an important role in glioma cell proliferation, apoptosis, and cell cycle.

Amplification and expression of a multidrug resistance gene in human glioma cell lines

1990 ◽  
Vol 72 (1) ◽  
pp. 96-101 ◽  
Author(s):  
Tsuyoshi Matsumoto ◽  
Eiichi Tani ◽  
Keizo Kaba ◽  
Nobuo Kochi ◽  
Hideki Shindo ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Multidrug Resistant ◽  
Glioma Cell Line ◽  
Cross Resistance ◽  
Human Glioma ◽  
Human Glioma Cell ◽  
Glioma Cell Lines

✓ Two human glioma cell lines were examined for multidrug resistance (MDR). A vincristine (VCR)-resistant glioma cell line showed a cross resistance to Adriamycin (doxorubicin, ADR) and etoposide (VP-16) to varying extents, suggesting the presence of MDR; the resistance to VCR was considerably decreased by calcium entry blockers. On the other hand, another VCR-sensitive glioma cell line exhibited no cross resistance to ADR or VP-16. Double minute chromosomes and homogeneously staining regions as well as clonal aberrations of chromosome 7 were not observed in cytogenetic studies of multidrug-resistant and multidrug-sensitive glioma cell lines. In Northern and Southern blot analyses, MDR gene 1 (MDR1) messenger ribonucleic acid (mRNA) was shown to be overexpressed without any amplification of the MDR1 gene in multidrug-resistant glioma cell lines as compared to multidrug-sensitive glioma cell lines. It would be reasonable to suggest that amplification of the MDR1 gene may not be a sine qua non for acquisition of MDR and that the MDR1 mRNA level may be well correlated with the extent of MDR.

scholarly journals Comparison of 99mTc-Tetrofosmin and 99mTc-Sestamibi Uptake in Glioma Cell Lines: The Role of P-Glycoprotein Expression

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
George A. Alexiou ◽  
Xanthi Xourgia ◽  
Evrysthenis Vartholomatos ◽  
Spyridon Tsiouris ◽  
John A. Kalef-Ezra ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Tumor Imaging ◽  
High Grade Glioma ◽  
High Grade ◽  
P Glycoprotein ◽  
99Mtc Tetrofosmin ◽  
Glioma Cell Lines

Tc-Tetrofosmin (Tc-TF) and Tc-Sestamibi (Tc-MIBI) are SPECT tracers that have been used for brain tumor imaging. Tumor’s multidrug resistance phenotype, namely, P-glycoprotein (p-gp), and the multidrug resistance related proteins (MRPs) expression have been suggested to influence both tracers’ uptake. In the present study we set out to compare Tc-MIBI uptake in high-grade glioma cell lines and to investigate the influence of gliomas p-gp expression on both tracers’ uptake. We used four glioma cell lines (U251MG, A172, U87MG, and T98G). The expression of p-gp protein was evaluated by flow cytometry. Twenty μCi (7.4·105 Bq) of Tc-TF and Tc-MIBI were used. The radioactivity in the cellular lysate was measured with a dose calibrator. P-gp was significantly expressed only in the U251MG cell line (). In all gliomas cell lines (U251MG, U87MG, A172, and T98G) the Tc-TF uptake was significantly higher than Tc-sestamibi. The U251MG cell line, in which significant p-gp expression was documented, exhibited the strongest uptake difference. Tc-TF uptake was higher than Tc-MIBI in all studied high-grade glioma cell lines. Thus, Tc-TF may be superior to Tc-MIBI for glioma imaging in vivo.

Receptor expression, cytogenetic, and molecular analysis of six continuous human glioma cell lines

1998 ◽  
Vol 34 (6) ◽  
pp. 455-462 ◽  
Author(s):  
Carol A. Kruse ◽  
Marileila Varella-Garcia ◽  
Bette K. Kleinschmidt-Demasters ◽  
Geoffrey C. Owens ◽  
Elaine B. Spector ◽  
...  
Keyword(s):  
Cell Lines ◽  
Molecular Analysis ◽  
Glioma Cell ◽  
Receptor Expression ◽  
Human Glioma ◽  
Human Glioma Cell ◽  
Glioma Cell Lines

Induction of autophagic cell death and radiosensitization by the pharmacological inhibition of nuclear factor–kappa B activation in human glioma cell lines

2009 ◽  
Vol 110 (3) ◽  
pp. 594-604 ◽  
Author(s):  
Yoshifumi Tsuboi ◽  
Masanori Kurimoto ◽  
Shoichi Nagai ◽  
Yumiko Hayakawa ◽  
Hironaga Kamiyama ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Nuclear Factor Kappa B ◽  
Nuclear Factor ◽  
Human Glioma ◽  
Pharmacological Inhibition ◽  
Glioma Cell Lines ◽  
Radiation Induced

Object The intrinsic radioresistance of certain cancer cells may be closely associated with the constitutive activation of nuclear factor–kappa B (NF-κB) activity, which may lead to protection from apoptosis. Recently, nonapoptotic cell death, or autophagy, has been revealed as a novel response of cancer cells to ionizing radiation. In the present study, the authors analyzed the effect of pitavastatin as a potential inhibitor of NF-κB activation on the radiosensitivity of A172, U87, and U251 human glioma cell lines. Methods The pharmacological inhibition of NF-κB activation was achieved using pitavastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Growth and radiosensitivity assays were performed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Hoechst 33258 staining, supravital acridine orange staining, and electron microscopy were performed utilizing 3 glioma cell lines with or without pitavastatin pretreatment to identify apoptosis or autophagy after irradiation. Results The growth of these 3 glioma cell lines was not significantly inhibited by pitavastatin at a concentration of up to 1 μM. Treatment with 0.1 μM of pitavastatin enhanced radiation-induced cell death in all glioma cell lines, with different sensitivity. Apoptosis did not occur in any pretreated or untreated (no pitavastatin) cell line following irradiation. Instead, autophagic cell changes were observed regardless of the radiosensitivity of the cell line. An inhibitor of autophagy, 3-methyladenine suppressed the cytotoxic effect of irradiation with pitavastatin, indicating that autophagy is a result of an antitumor mechanism. Using the most radiosensitive A172 cell line, the intracellular localization of p50, a representative subunit of NF-κB, was evaluated through immunoblotting and immunofluorescence studies. The NF-κB of A172 cells was immediately activated and translocated from the cytosol to the nucleus in response to irradiation. Pitavastatin inhibited this activation and translocation of NF-κB. Conclusions Autophagic cell death rather than apoptosis is a possible mechanism of radiation-induced and pitavastatin-enhanced cell damage, and radiosensitization by the pharmacological inhibition of NF-κB activation may be a novel therapeutic strategy for malignant gliomas.

scholarly journals ATRX loss induces multiple hallmarks of the alternative lengthening of telomeres (ALT) phenotype in human glioma cell lines in a cell line-specific manner

PLoS ONE ◽  
2018 ◽  
Vol 13 (9) ◽  
pp. e0204159 ◽  
Author(s):  
Jacqueline A. Brosnan-Cashman ◽  
Ming Yuan ◽  
Mindy K. Graham ◽  
Anthony J. Rizzo ◽  
Kaylar M. Myers ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Human Glioma ◽  
Alternative Lengthening Of Telomeres ◽  
Human Glioma Cell ◽  
Alternative Lengthening ◽  
Glioma Cell Lines ◽  
Specific Manner ◽  
A Cell

scholarly journals Multiple Localization of Endogenous MARK4L Protein in Human Glioma

2009 ◽  
Vol 31 (5) ◽  
pp. 357-370
Author(s):  
Ivana Magnani ◽  
Chiara Novielli ◽  
Melissa Bellini ◽  
Gaia Roversi ◽  
Lorenzo Bello ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Cellular Localization ◽  
Protein Identification ◽  
Glioblastoma Cell Line ◽  
Glioblastoma Cell ◽  
Human Glioma ◽  
Glioma Cell Lines ◽  
Biochemical Fractionation

Background: We have previously shown that the sustained expression of MARK4L transcripts in glioma and neural progenitors (NHNPs) declines after exposure to antisense MARK4L oligonucleotides in glioblastoma cell lines. Array-CGH confirmed the genomic duplication of MARK4L identified by FISH in a glioblastoma cell line. This background together with literature data on the exogenous association of MARK4 with interphase centrosome prompted us to investigate the sub-cellular localization of the endogenous MARK4L protein aiming at achieving insights on its possible role in the pathomechanisms of glioma.Methods: Immunodetection was carried out to validate the specificity of MARK4L antibody in gliomas and NHNPs. Mass spectrometry was applied for MARK4L protein identification in a representative glioblastoma cell line. Combined biochemical fractionation and immunodetection analyses were performed to confirm the sub-cellular localization of MARK4L achieved by immunofluorescence in glioma cell lines.Results: By assigning MARK4L protein within the band immunoprecipitated by the specific antibody we validated our anti-MARK4L antibody. We demonstrated that the endogenous MARK4L: (i) colocalizes with centrosomes at all mitotic stages and resides in centrosome-enriched fractions; (ii) associates with the nucleolus and the midbody and respective fractions, and (iii) co-stains the aberrant centrosome configurations observed in glioma cell lines.Conclusions: The overall data merge on the multiplex entry of MARK4L into the cell cycle and link it to the aberrant centrosomes in glioma cell lines suggesting a possible role of this kinase in the abnormal mitotic processes of human glioma.

scholarly journals Temozolomide sensitivity of malignant glioma cell lines: a systematic review assessing consistencies between in vitro studies

2021 ◽  
Author(s):  
Michael TC Poon ◽  
Morgan Bruce ◽  
Joanne Simpson ◽  
Cathal J Hannan ◽  
Paul M Brennan
Keyword(s):  
Cell Viability ◽  
Malignant Glioma ◽  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Glioma Cell Line ◽  
In Vitro Studies ◽  
Experimental Conditions ◽  
Glioma Cell Lines

Background: Malignant glioma cell line models are integral to pre-clinical testing of novel potential therapies. Accurate prediction of likely efficacy in the clinic requires that these models are reliable and consistent. We assessed this by examining the reporting of experimental conditions and sensitivity to temozolomide in glioma cells lines. Methods: We searched Medline and Embase (Jan 1994-Jan 2021) for studies that evaluated the effect of temozolomide monotherapy on cell viability of at least one malignant glioma cell line. Studies using a drug-resistant cell line or a modified preparation of temozolomide were excluded. Key data items included type of cell lines, temozolomide exposure duration, and cell viability measure (IC50). Results: We included 212 eligible studies from 2,789 non-duplicate records that reported 248 distinct cell lines. The commonest cell line was U87 (60.4%). Only 10.4% studies used a patient-derived cell line. The proportion of studies not reporting each experimental condition ranged from 8.0-27.4%, including base medium (8.0%), serum supplementation (9.9%) and number of replicates (27.4%). In studies reporting IC50 the median value for U87 cell line at 24 hours, 48 hours and 72 hours was 123.9μM (IQR 75.3-277.7μM), 223.1μM (IQR 92.0-590.1μM) and 230.0μM (IQR 34.1-650.0μM), respectively (Figure 2A). The median IC50 at 72 hours for patient-derived cell lines was 220μM (IQR 81.1-800.0μM). Conclusions: Temozolomide sensitivity reported in comparable studies was not consistent between and within individual malignant glioma cell lines. Drug discovery science performed on these models cannot reliably inform clinical translation. A consensus model of reporting can maximise reproducibility and consistency among in vitro studies.

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