Genetic Analysis of a Multifocal Glioblastoma Multiforme: A Suitable Tool to Gain New Aspects in Glioma Development

Neurosurgery ◽  
2003 ◽  
Vol 53 (6) ◽  
pp. 1377-1384 ◽  
Author(s):  
Dietmar Krex ◽  
Brigitte Mohr ◽  
Hella Appelt ◽  
Hans K. Schackert ◽  
Gabriele Schackert
Keyword(s):  
Glioblastoma Multiforme ◽  
Loss Of Heterozygosity ◽  
Cell Lines ◽  
Homozygous Deletion ◽  
Receptor Protein ◽  
Pten Gene ◽  
Balanced Translocation ◽  
In Vitro Proliferation ◽  
Suitable Tool

Abstract OBJECTIVE Multifocal glioblastomas constitute an increasingly diagnosed subgroup of glioblastoma multiforme, the most malignant primary brain tumor in adults. The molecular background of these lesions is unknown. However, the ability to study multiple lesions of one patient simultaneously could provide new aspects in glioma development. METHODS Short-term cell cultures were derived from three isolated glioblastoma lesions of one patient. Spectral karyotyping and interphase fluorescence in situ hybridization were used for cytogenetic analysis. Loss of heterozygosity was assessed in tumor tissues and cell lines for seven gene loci (p73, p21, p16, PTEN, p27, Rb, p53). In addition, sequence analysis of the PTEN and p53 loci was performed, epidermal growth factor receptor protein expression was assessed, and in vitro proliferation was assayed. RESULTS A balanced translocation [t(1;15)(p3?6;q2?5)] that has not been described previously in glioblastomas was identified in all cell lines. Primarily, the cell lines have a homozygous deletion of the p16 locus and inactivation of the PTEN gene by loss of heterozygosity and an identical mutation in common. Furthermore, the cell lines harbor a hemizygous (R175H) or two heterozygous (R175H, R213Q) mutations of the p53 gene or none at all. The occurrence of p53 mutations correlates with the size of the original tumors and in vitro proliferation. CONCLUSION The analysis of a multifocal glioma revealed three main aspects: 1) the combined cytogenetic and molecular analysis of this subgroup of glioblastoma multiforme is a suitable tool to gain new perspectives in glioma development, 2) the balanced translocation [t(1;15)(p3?6;q2?5)] might harbor a new genetic marker involved in glioma development, and 3) the pattern of p53 mutation suggests a role of p53 in the progression of malignancy, migration, and growth of this particular primary glioblastoma.

scholarly journals Lomeguatrib Increases the Radiosensitivity of MGMT Unmethylated Human Glioblastoma Multiforme Cell Lines

2021 ◽  
Vol 22 (13) ◽  
pp. 6781
Author(s):  
Anna Kirstein ◽  
Daniela Schilling ◽  
Stephanie E. Combs ◽  
Thomas E. Schmid
Keyword(s):  
Glioblastoma Multiforme ◽  
Cell Lines ◽  
Dna Methyltransferase ◽  
Treatment Options ◽  
Treatment Resistance ◽  
Cell Cycle Distribution ◽  
Human Glioblastoma ◽  
Human Glioblastoma Multiforme ◽  
Mgmt Protein

Background: Treatment resistance of glioblastoma multiforme to chemo- and radiotherapy remains a challenge yet to overcome. In particular, the O6-methylguanine-DNA-methyltransferase (MGMT) promoter unmethylated patients have only little benefit from chemotherapy treatment using temozolomide since MGMT counteracts its therapeutic efficacy. Therefore, new treatment options in radiotherapy need to be developed to inhibit MGMT and increase radiotherapy response. Methods: Lomeguatrib, a highly specific MGMT inhibitor, was used to inactivate MGMT protein in vitro. Radiosensitivity of established human glioblastoma multiforme cell lines in combination with lomeguatrib was investigated using the clonogenic survival assay. Inhibition of MGMT was analyzed using Western Blot. Cell cycle distribution and apoptosis were investigated to determine the effects of lomeguatrib alone as well as in combination with ionizing radiation. Results: Lomeguatrib significantly decreased MGMT protein and reduced radiation-induced G2/M arrest. A radiosensitizing effect of lomeguatrib was observed when administered at 1 µM and increased radioresistance at 20 µM. Conclusion: Low concentrations of lomeguatrib elicit radiosensitization, while high concentrations mediate a radioprotective effect.

scholarly journals TGF-β Promotes the Proliferation of Microglia In Vitro

2019 ◽  
Vol 10 (1) ◽  
pp. 20 ◽  
Author(s):  
Costansia Bureta ◽  
Takao Setoguchi ◽  
Yoshinobu Saitoh ◽  
Hiroyuki Tominaga ◽  
Shingo Maeda ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Lines ◽  
Transforming Growth Factor Beta ◽  
Microglial Cell ◽  
Transforming Growth Factor ◽  
Dependent Manner ◽  
In Vitro Proliferation ◽  
Inhibition Of Proliferation ◽  
Significant Difference

The activation and proliferation of microglia is characteristic of the early stages of brain pathologies. In this study, we aimed to identify a factor that promotes microglial activation and proliferation and examined the in vitro effects on these processes. We cultured microglial cell lines, EOC 2 and SIM-A9, with various growth factors and evaluated cell proliferation, death, and viability. The results showed that only transforming growth factor beta (TGF-β) caused an increase in the in vitro proliferation of both microglial cell lines. It has been reported that colony-stimulating factor 1 promotes the proliferation of microglia, while TGF-β promotes both proliferation and inhibition of cell death of microglia. However, upon comparing the most effective doses of both (assessed from the proliferation assay), we identified no statistically significant difference between the two factors in terms of cell death; thus, both have a proliferative effect on microglial cells. In addition, a TGF-β receptor 1 inhibitor, galunisertib, caused marked inhibition of proliferation in a dose-dependent manner, indicating that inhibition of TGF-β signalling reduces the proliferation of microglia. Therefore, galunisertib may represent a promising therapeutic agent for the treatment of neurodegenerative diseases via inhibition of nerve injury-induced microglial proliferation, which may result in reduced inflammatory and neuropathic and cancer pain.

scholarly journals C-Phycocyanin Suppresses the In Vitro Proliferation and Migration of Non-Small-Cell Lung Cancer Cells through Reduction of RIPK1/NF-κB Activity

Marine Drugs ◽  
2019 ◽  
Vol 17 (6) ◽  
pp. 362 ◽  
Author(s):  
Shuai Hao ◽  
Shuang Li ◽  
Jing Wang ◽  
Lei Zhao ◽  
Yan Yan ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Cell Lines ◽  
Cell Lung Cancer ◽  
Small Cell ◽  
Cell Phenotype ◽  
Small Cell Lung ◽  
In Vitro Proliferation ◽  
And Migration

Phycocyanin, derived from Spirulina platensis, is a type of natural antineoplastic marine protein. It is known that phycocyanin exerts anticancer effects on non-small-cell lung cancer (NSCLC) cells, but its underlying mechanism has not been elucidated. Herein, the antitumor function and regulatory mechanism of phycocyanin were investigated in three NSCLC cell lines for the first time: H358, H1650, and LTEP-a2. Cell phenotype experiments suggested that phycocyanin could suppress the survival rate, proliferation, colony formation, and migration abilities, as well as induce apoptosis of NSCLC cells. Subsequently, transcriptome analysis revealed that receptor-interacting serine/threonine-protein kinase 1 (RIPK1) was significantly down-regulated by phycocyanin in the LTEP-a2 cell, which was further validated by qRT-PCR and Western blot analysis in two other cell lines. Interestingly, similar to phycocyanin-treated assays, siRNA knockdown of RIPK1 expression also resulted in growth and migration inhibition of NSCLC cells. Moreover, the activity of NF-κB signaling was also suppressed after silencing RIPK1 expression, indicating that phycocyanin exerted anti-proliferative and anti-migratory function through down-regulating RIPK1/NF-κB activity in NSCLC cells. This study proposes a mechanism of action for phycocyanin involving both NSCLC apoptosis and down regulation of NSCLC genes.

Isolation of immortalized, INK4a/ARF-deficient cells from the subventricular zone after in utero N-ethyl-N-nitrosourea exposure

2005 ◽  
Vol 102 (1) ◽  
pp. 98-108 ◽  
Author(s):  
Todd M. Savarese ◽  
Taichang Jang ◽  
Hoi Pang Low ◽  
Rebecca Salmonsen ◽  
N. Scott Litofsky ◽  
...  
Keyword(s):  
Cell Lines ◽  
Subventricular Zone ◽  
Defined Medium ◽  
In Utero ◽  
Homozygous Deletion ◽  
Content Type ◽  
Immortalized Cell Lines ◽  
Immortalized Cell ◽  
Fine Print

Object. Brain tumors, including gliomas, develop several months after rats are exposed in utero to N-ethyl-N-nitrosourea (ENU). Although pathological changes cannot be detected until these animals are several weeks old, the process that eventually leads to glioma formation must begin soon after exposure given the rapid clearance of the carcinogen and the observation that transformation of brain cells isolated soon after exposure occasionally occurs. This model can therefore potentially provide useful insights about the early events that precede overt glioma formation. The authors hypothesized that future glioma cells arise from stem/progenitor cells residing in or near the subventricular zone (SVZ) of the brain. Methods. Cells obtained from the SVZ or corpus striatum in ENU-exposed and control rats were cultured in an epidermal growth factor (EGF)-containing, chemically defined medium. Usually, rat SVZ cells cultured in this manner (neurospheres) are nestin-positive, undifferentiated, and EGF-dependent and undergo cell senescence. Consistent with these prior observations, control SVZ cells undergo senescence by the 12th to 15th doubling (20 of 20 cultures). In contrast, three of 15 cultures of cells derived from the SVZs of individual ENU-treated rats continue to proliferate for more than 60 cell passages. Each of these nestin-expressing immortalized cell lines harbored a common homozygous deletion spanning the INK4a/ARF locus and was unable to differentiate into neural lineages after exposure to specific in vitro stimuli. Nevertheless, unlike the rat C6 glioma cell line, these immortalized cell lines demonstrate EGF dependence and low clonogenicity in soft agar and did not form tumors after intracranial transplantation. Conclusions. Data in this study indicated that immortalized cells may represent glioma precursors that reside in the area of the SVZ after ENU exposure that may serve as a reservoir for further genetic and epigenetic hits that could eventually result in a full glioma phenotype.

Phosphodiesterase (PDE) inhibitors reduce in vitro proliferation of prostate primary cells but do not interfere with growth of prostate carcinoma cell lines

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 14604-14604 ◽  
Author(s):  
D. G. Pernkopf ◽  
G. Untergasser ◽  
P. Berger ◽  
E. Plas
Keyword(s):  
Cell Lines ◽  
Prostate Carcinoma ◽  
Carcinoma Cell ◽  
Polyclonal Antiserum ◽  
Primary Cells ◽  
In Vitro Proliferation ◽  
Carcinoma Cell Lines ◽  
Prostate Carcinoma Cell ◽  
High Concentrations

14604 Background: PDE 5 is highly expressed in cavernosal and prostatic tissue. The mechanism of PDE-5 inhibitors on cavernosal tissue is well established but the effects of PDE-5 inhibitors on prostatic cells are unknown. The aim of this study was to analyze in vitro effects of PDE-5 inhibitors on prostate primary cells, fibroblasts (PrSC), basal epithelial cells (PrEC) and prostate cancer cell lines. Methods: Cultivated PrEC and PrSC, immortalized BPH cells (BPH 1), androgen dependent (LNCaP) and androgen independent (PC3) prostate carcinoma cell lines were exposed to increasing concentrations of commercially available PDE 5 inhibitors Sildenafil (Sil), Tadalafil (Tad), Vardenafil (Var). After incubation for 3 days cell viability was determined by a WST-1 assay (Roche-Biochemicals). Cells were evaluated morphologically by invert-light microscopy. PDE-5 protein concentrations were determined by western blot analyses and tissue distribution of PDE-5 by immunohistochemistry (IHC) with a polyclonal antiserum. Results: None of the PDE-5 inhibitors induced cell proliferation. Significant decreases in proliferation and viability were observed at high concentrations (1 mg/ml) of all substances in PrSC and PrEC. In PrSC, proliferation rate decreased to 37.7% in Sil, to 16.9% in Tad and to 63.7% for Var as compared to controls. In PrEC, proliferation decreased to 72.7%, 21.6% and 84.4% for Sil, Tad and Var, respectively. At 0.1 mg/ml only Tad reduced proliferation significantly to 57.4%. Moreover, Tad induced neuroendocrine-like morphology in some PrEC. High protein concentrations of PDE 5 were observed in PrEC, low concentrations in PrSC but none in cancer cells. Conclusions: Sil, Tad and Var inhibit proliferation of prostate primary cells in vitro. Tad showed highest inhibition. Tumor cells were insensitive to PDE-5 inhibitors, due to the lack of PDE-5 protein. It seems unlikely that any of these substances increases proliferation of prostate carcinoma. Tad induced neuroendocrine-like morphology in some basal PrEC indicating effects on cellular differentiation. No significant financial relationships to disclose.

scholarly journals Infectious Mononucleosis: In Vitro Evidence for Limited Lymphoproliferation

Blood ◽  
1969 ◽  
Vol 33 (2) ◽  
pp. 292-299 ◽  
Author(s):  
PHILIP R. GLADE ◽  
YASHAR HIRSHAUT ◽  
DANIEL P. STITES ◽  
LAWRENCE N. CHESSIN
Keyword(s):  
Cell Lines ◽  
Infectious Mononucleosis ◽  
Normal State ◽  
Peripheral Blood ◽  
Close Correlation ◽  
Transient Phenomenon ◽  
In Vitro Proliferation ◽  
Laboratory Parameters

Abstract The persistence of circulating leukocytes with potential for long-term in vitro proliferation was investigated in patients with heterophile-positive infectious mononucleosis. The ability to isolate long-term suspension cultures from peripheral blood with selective technics was a transient phenomenon for each patient studied, disappearing with a return of clinical and laboratory parameters toward the normal state. There was no close correlation between the ability to isolate cell lines and clinical, morphologic, biosynthetic or serologic manifestations of disease.

In vitro efficacy of transferrin-toxin conjugates against glioblastoma multiforme

1992 ◽  
Vol 76 (5) ◽  
pp. 838-844 ◽  
Author(s):  
Walter A. Hall ◽  
Aslak Godal ◽  
Siri Juell ◽  
Øystein Fodstad
Keyword(s):  
Glioblastoma Multiforme ◽  
Cytotoxic Activity ◽  
Cell Lines ◽  
Receptor Expression ◽  
Binding Assays ◽  
Content Type ◽  
Monensin A ◽  
Diferric Transferrin

✓ The cytotoxic activity of immunotoxins constructed with human diferric transferrin (Tfn) as the carrier ligand and an abrin variant Pseudomonas exotoxin A (PE) and the diphtheria toxin mutant cross-reacting material (CRM) 107 as the toxin moieties were studied in vitro. Three malignant human cell lines, the glioblastomas multiforme SNB19 and SF295 and the LOX melanoma, and a nonhuman control murine melanoma cell line B16 were assessed. The presence of transferrin receptors on the cell lines was confirmed by direct 125I-Tfn binding assays. The 50% protein synthesis inhibitory concentration (IC50) values for all cell lines demonstrated that Tfn-abrin variant and Tfn-PE had comparable potency and were both more effective than Tfn-CRM 107. Monensin, a carboxylic ionophore, potentiated the effect of Tfn-abrin variant against glioma cells approximately 35-fold with IC50 values of 4.0 × 10−13 M and 4.7 × 10−12 M for SNB19 and SF295, respectively. Cytotoxic activity of Tfn-abrin variant (with or without monensin) and Tfn-PE was correlated with the degree of Tfn receptor expression measured on the cell lines. The exquisite in vitro cytotoxicity of Tfn-abrin variant and Tfn-PE immunotoxins against glioma and melanoma cells warrants further in vivo evaluation and future consideration of these agents for potential clinical application against glioblastoma multiforme and leptomeningeal neoplasia.

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