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.

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.

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

Upregulation of miRNA-202 promotes apoptosis and inhibits migration of glioma cell line via downregulation of ROCK1 gene

2020 ◽  
Author(s):  
Mousa Behzadi ◽  
Hamed Hatami ◽  
Fatemeh Alian ◽  
Maryam Shojaee ◽  
Masoumeh Alimohammadi ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Glioma Cell ◽  
Glioma Cell Line ◽  
Human Glioma ◽  
Human Glioma Cell Line ◽  
Human Glioma Cell ◽  
Glioma Cell Lines ◽  
And Migration ◽  
Line P

Abstract Background: We evaluated role(s) of miR-202 in glioma cell lines, its effect on ROCK1 expression, and also evaluation of apoptosis and migration of human glioma cell line after transfection with miR-202 mimics and inhibitors. Material and methods: The cell lines were transfected with mimic, inhibitor and NC of miR-202. Reverse transcription polymerase chain reaction (RT-PCR) was conducted to evaluate the expression of miR‐202 and ROCK1 . Western blot was performed to detect the protein level of ROCK1. Furthermore, MTT and wound healing assay were performed to evaluate the effects of miR-202 on apoptosis and migration of human glioma cell line, respectively. Results: miR-202 showed a significantly decrease in human glioma cell lines, compared with the NHA cell line (P<0.05). The ROCK1 expression was significantly upregulated in glioma cell lines, compared with the NHA cell line ( P <0.05). Furthermore, a negative correlation was observed between expression of ROCK1 and miR-202 ( P =0.01, r=-0.426). The mRNA and protein levels of ROCK1 were decreased in U87 cell line in miR-202 mimics group, compared with mimic NC group (P<0.05). In addition, apoptosis was significantly increased in miR-202 mimics, compared with the NC group in U87 cell line at 72 and 96 h (P<0.05). Furthermore, invasion showed a significant decrease in miR-202 mimic group, compared with U87 cell line at 24 and 48 h (P<0.05). Conclusions: The miR-202 could serve as a tumour-suppressor miRNA in glioma. Therefore, targeting ROCK1 by miR-202 may increase improve disease outcome and could be considered as a potential therapeutic target for glioma patients.

Expression of P-glycoprotein in human glioma cell lines and surgical glioma specimens

1991 ◽  
Vol 74 (3) ◽  
pp. 460-466 ◽  
Author(s):  
Tsuyoshi Matsumoto ◽  
Eiichi Tani ◽  
Keizo Kaba ◽  
Hideki Shindo ◽  
Katsuya Miyaji
Keyword(s):  
Multidrug Resistance ◽  
Cell Line ◽  
Cell Lines ◽  
Western Blotting ◽  
Glioma Cell ◽  
Human Glioma ◽  
Content Type ◽  
P Glycoprotein ◽  
Glioma Cell Lines ◽  
Fine Print

✓ The expression of P-glycoprotein, a product of multidrug resistance gene 1, was studied by Western blotting and immunohistochemistry in five human glioma cell lines. One glioma cell line was resistant to vincristine, Adriamycin (doxorubicin), and etoposide, and the other four glioma cell lines were sensitive to each drug. The multidrug-resistant cell line showed a high expression of P-glycoprotein in Western blot analysis and a positive immunostaining for P-glycoprotein mainly along the cell membrane, whereas all multidrug-sensitive glioma cell lines demonstrated no expression of P-glycoprotein in Western blotting and no immunostaining for P-glycoprotein, thus showing a good correlation between the expression level of P-glycoprotein and the extent of multidrug resistance. In 18 human surgical glioma specimens, there was no evidence of complete absence of immunostaining for P-glycoprotein. With a definition of more than 20% of P-glycoprotein-positive cells as positive, from 10% to 20% as intermediate, and less than 10% as negative, immunostaining for P-glycoprotein was positive in one specimen and intermediate in six of 15 specimens taken from virgin gliomas, and positive in two specimens and intermediate in one of three recurrent gliomas treated previously with irradiation, ACNU (1-(4-amino-2-methyl-pyrimidine-5-yl)-methyl-3-(2-chloroethyl)-3-nitrosourea hydrochloride), cisplatin, vincristine, and/or procarbazine.

Enhancement of temozolomide-induced apoptosis by valproic acid in human glioma cell lines through redox regulation

2011 ◽  
Vol 89 (3) ◽  
pp. 303-315 ◽  
Author(s):  
Ching-Hsein Chen ◽  
Yu-Jia Chang ◽  
Maurice S. B. Ku ◽  
King-Thom Chung ◽  
Jen-Tsung Yang
Keyword(s):  
Valproic Acid ◽  
Cell Lines ◽  
Glioma Cell ◽  
Redox Regulation ◽  
Human Glioma ◽  
Human Glioma Cell ◽  
Glioma Cell Lines ◽  
Induced Apoptosis

scholarly journals Verotoxin-1 induction of apoptosis in Gb3-expressing human glioma cell lines

2006 ◽  
Vol 5 (9) ◽  
pp. 1211-1217 ◽  
Author(s):  
David Johansson ◽  
Anders Johansson ◽  
Kjell Grankvist ◽  
Ulrika Andersson ◽  
Roger Henriksson ◽  
...  
Keyword(s):  
Cell Lines ◽  
Glioma Cell ◽  
Human Glioma ◽  
Human Glioma Cell ◽  
Glioma Cell Lines ◽  
Induction Of Apoptosis

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.

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