Role of Luteolin-Induced Apoptosis and Autophagy in Human Glioblastoma Cell Lines

Background and Objectives: Malignant glioblastoma (GBM) is caused by abnormal proliferation of glial cells, which are found in the brain. The therapeutic effects of surgical treatment, radiation therapy, and chemo-therapy against GBM are relatively poor compared with their effects against other tumors. Luteolin is abundant in peanut shells and is also found in herbs and other plants, such as thyme, green pepper, and celery. Luteolin is known to be effective against obesity and metabolic syndrome. The anti-inflammatory, and anti-cancer activities of luteolin have been investigated. Most studies have focused on the antioxidant and anti-inflammatory effects of luteolin, which is a natural flavonoid. However, the association between the induction of apoptosis by luteolin in GBM and autophagy has not yet been investigated. This study thus aimed to confirm the occurrence of luteolin-induced apoptosis and autophagy in GBM cells and to assess their relationship. Materials and Methods: A172 and U-373MG glioblastoma cell lines were used for this experiment. We confirmed the apoptosis effect of Luteolin on GBM cells using methods such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, immunofluorescence, Flow cytometry (FACS) western blot, and real-time quantitative PCR (qPCR). Results: In the luteolin-treated A172 and U-373MG cells, cell viability decreased in a concentration- and time-dependent manner. In addition, in A172 and U-373MG cells treated with luteolin at concentrations greater than 100 μM, nuclear fragmentation, which is a typical morphological change characterizing apoptosis, as well as fragmentation of caspase-3 and Poly (ADP-ribose) polymerase (PARP), which are apoptosis-related factors, were observed. Autophagy was induced after treatment with at least 50 μM luteolin. Inhibition of autophagy using 3MA allowed for a low concentration of luteolin to more effectively induce apoptosis in A172 and U-373MG cells. Conclusions: Results showed that luteolin induces apoptosis and autophagy and that the luteolin-induced autophagy promotes cell survival. Therefore, an appropriate combination therapy involving luteolin and an autophagy inhibitor is expected to improve the prognosis of GBM treatment.

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Synthetic Smac peptide enhances the effect of etoposide-induced apoptosis in human glioblastoma cell lines

Journal of Neuro-Oncology ◽

10.1007/s11060-005-9045-5 ◽

2006 ◽

Vol 77 (3) ◽

pp. 247-255 ◽

Cited By ~ 21

Author(s):

Katsu Mizukawa ◽

Atsufumi Kawamura ◽

Takashi Sasayama ◽

Kazuhiro Tanaka ◽

Masahito Kamei ◽

...

Keyword(s):

Cell Lines ◽

Glioblastoma Cell ◽

Human Glioblastoma ◽

Human Glioblastoma Cell ◽

Induced Apoptosis ◽

Glioblastoma Cell Lines

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Targeting of CD317 by the immunotoxin HM1.24-ETA’ to allow immunotherapy in glioblastoma patients.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e13560 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. e13560-e13560

Author(s):

Dorothee Gramatzki ◽

Emese Szabo ◽

Martin Gramatzki ◽

Matthias Peipp ◽

Michael Weller

Keyword(s):

Mrna Expression ◽

Cell Lines ◽

Dependent Manner ◽

Glioblastoma Cell ◽

Single Chain ◽

Human Glioblastoma ◽

Human Glioblastoma Cell ◽

Protein Levels ◽

Glioblastoma Cell Lines

e13560 Background: Glioblastoma is the most common primary malignant brain tumor with a poor prognosis. CD317 (HM1.24) is a transmembrane protein and may exist in differently spliced variants. It is highly expressed on plasma cells in multiple myeloma, as well as in certain solid tumor types. While several antibody drug conjugates are already in clinical practice, small immunotoxins with a different intracellular mode of action are only established in hairy cell leukemia. The immunotoxin HM1.24-ETA’ protein is a CD317 single chain Fv (scFv) antibody fused to a truncated version of Pseudomonas aeruginosa exotoxin A (ETA’). Methods: In vivo CD317 mRNA expression in human glioma of different grades and survival probabilities of glioblastoma patients based on CD317 mRNA expression were analyzed using the database of the Cancer Genome Atlas network (TCGA). CD317 protein expression was analyzed by immunohistochemistry in a human tissue microarray (TMA). In vitro CD317 mRNA expression was assessed by RT-PCR and CD317 protein levels by flow cytometry in several human glioblastoma cell lines. A cytotoxicity assay after treatment with HM1.24-ETA’ immunotoxin was performed in human glioblastoma cell lines. Results: Data on mRNA expression from the TCGA database demonstrated, that CD317 was upregulated in human glioblastomas compared to lower grade gliomas. In the group of glioblastoma patients increased CD317 mRNA expression was associated with decreased probability of survival ( p< 0.001). CD317 protein levels correlated directly with the tumor grade of astrocytic gliomas in the TMA. CD317 was expressed heterogeneously on mRNA and protein levels in the tested cell-lines in vitro. HM1.24-ETA’ induced cytotoxicity in CD317-positive glioblastoma cells in a concentration-dependent manner. Animal experiments currently performed suggest activity in glioblastoma xenografted mice. Conclusions: These data highlight CD317 as an interesting target antigen and HM1.24-ETA’ immunotoxin as a strategy for immunotherapy of glioblastoma patients.

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The lipid phosphatase activity of PTEN is critical for stabilizing intercellular junctions and reverting invasiveness

The Journal of Cell Biology ◽

10.1083/jcb.200105109 ◽

2001 ◽

Vol 155 (7) ◽

pp. 1129-1136 ◽

Cited By ~ 74

Author(s):

Larissa Kotelevets ◽

Jolanda van Hengel ◽

Erik Bruyneel ◽

Marc Mareel ◽

Frans van Roy ◽

...

Keyword(s):

Cell Line ◽

Phosphatase Activity ◽

Cell Lines ◽

Dependent Manner ◽

Glioblastoma Cell ◽

Wild Type ◽

Invasive Phenotype ◽

Lipid Phosphatase ◽

Glioblastoma Cell Lines ◽

E Cadherin

To analyze the implication of PTEN in the control of tumor cell invasiveness, the canine kidney epithelial cell lines MDCKras-f and MDCKts-src, expressing activated Ras and a temperature-sensitive v-Src tyrosine kinase, respectively, were transfected with PTEN expression vectors. Likewise, the human PTEN-defective glioblastoma cell lines U87MG and U373MG, the melanoma cell line FM-45, and the prostate carcinoma cell line PC-3 were transfected. We demonstrate that ectopic expression of wild-type PTEN in MDCKts-src cells, but not expression of PTEN mutants deficient in either the lipid or both the lipid and protein phosphatase activities, reverted the morphological transformation, induced cell–cell aggregation, and suppressed the invasive phenotype in an E-cadherin–dependent manner. In contrast, overexpression of wild-type PTEN did not counteract Ras-induced invasiveness of MDCKras-f cells expressing low levels of E-cadherin. PTEN effects were not associated with marked changes in accumulation or phosphorylation levels of E-cadherin and associated catenins. Wild-type, but not mutant, PTEN also reverted the invasive phenotype of U87MG, U373MG, PC-3, and FM-45 cells. Interestingly, PTEN effects were mimicked by N-cadherin–neutralizing antibody in the glioblastoma cell lines. Our data confirm the differential activities of E- and N-cadherin on invasiveness and suggest that the lipid phosphatase activity of PTEN exerts a critical role in stabilizing junctional complexes and restraining invasiveness.

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Combination MEK and mTOR inhibitor therapy is active in models of glioblastoma

Neuro-Oncology Advances ◽

10.1093/noajnl/vdaa138 ◽

2020 ◽

Vol 2 (1) ◽

Author(s):

Karisa C Schreck ◽

Amy N Allen ◽

Jiawan Wang ◽

Christine A Pratilas

Keyword(s):

Cell Lines ◽

Kinase Inhibitor ◽

De Novo ◽

Mek Inhibitor ◽

Glioblastoma Cell ◽

Mtor Inhibition ◽

Adaptive Resistance ◽

Downstream Signaling ◽

Induced Apoptosis ◽

Glioblastoma Cell Lines

Abstract Background RAS effector signaling pathways such as PI3K/mTOR and ERK are frequently dysregulated in glioblastoma. While small molecule targeted therapies against these pathways have appeared promising in preclinical studies, they have been disappointing in clinical trials due to toxicity and de novo and adaptive resistance. To identify predictors of glioblastoma sensitivity to dual pathway inhibition with mTORC1/2 and MEK inhibitors, we tested these agents, alone and in combination, in a cohort of genomically characterized glioblastoma cell lines. Methods Seven genomically characterized, patient-derived glioblastoma neurosphere cell lines were evaluated for their sensitivity to the dual mTORC1/2 kinase inhibitor sapanisertib (MLN0128, TAK-228) alone or in combination with the MEK1/2 inhibitor trametinib (GSK1120212), using assessment of proliferation and evaluation of the downstream signaling consequences of these inhibitors. Results Sapanisertib inhibited cell growth in neurosphere lines, but induced apoptosis only in a subset of lines, and did not completely inhibit downstream mTOR signaling via ribosomal protein S6 (RPS6). Growth sensitivity to MEK inhibitor monotherapy was observed in a subset of lines defined by loss of NF1, was predicted by an ERK-dependent expression signature, and was associated with effective phospho-RPS6 inhibition. In these lines, combined MEK/mTOR treatment further inhibited growth and induced apoptosis. Combined MEK and mTOR inhibition also led to modest antiproliferative effects in lines with intact NF1 and insensitivity to MEK inhibitor monotherapy. Conclusions These data demonstrate that combined MEK/mTOR inhibition is synergistic in glioblastoma cell lines and may be more potent in NF1-deficient glioblastoma.

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The inhibition of anti-inflammatory cytokines in glioblastoma cell lines by WEE1 gene silencing with specific ShRNA

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2011.08.150 ◽

2011 ◽

Vol 44 (13) ◽

pp. S70

Author(s):

Jaberipour Mansooreh ◽

Rahnama Susan ◽

Hosseini Ahmad ◽

Habibagahi Mojtaba

Keyword(s):

Gene Silencing ◽

Cell Lines ◽

Inflammatory Cytokines ◽

Glioblastoma Cell ◽

Anti Inflammatory ◽

Glioblastoma Cell Lines

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Synthetic Cannabinoids Influence the Invasion of Glioblastoma Cell Lines in a Cell- and Receptor-Dependent Manner.

Cancers ◽

10.3390/cancers11020161 ◽

2019 ◽

Vol 11 (2) ◽

pp. 161 ◽

Cited By ~ 3

Author(s):

Tim Hohmann ◽

Kerstin Feese ◽

Thomas Greither ◽

Chalid Ghadban ◽

Vivian Jäger ◽

...

Keyword(s):

Cell Lines ◽

Cannabinoid Receptor ◽

Synthetic Cannabinoids ◽

Current Treatment ◽

Current Therapy ◽

Dependent Manner ◽

Glioblastoma Cell ◽

Proliferation And Apoptosis ◽

A Cell ◽

Glioblastoma Cell Lines

The current treatment of glioblastoma is not sufficient, since they are heterogeneous and often resistant to chemotherapy. Earlier studies demonstrated effects of specific cannabinoid receptor (CB) agonists on the invasiveness of glioblastoma cell lines, but the exact mechanism remained unclear. Three human glioblastoma cell lines were treated with synthetic CB ligands. The effect of cannabinoids on microRNAs (miRs), Akt, and on the expression of proliferation and apoptosis markers were analyzed. Furthermore, in a model of organotypic hippocampal slice cultures cannabinoid mediated changes in the invasiveness were assessed. MicroRNAs and the activation of Akt which are related to cell migration, apoptosis, and proliferation were evaluated and found not to be associated with changes in the invasiveness after treatment with CB ligands. Also proliferation and/or apoptosis were not altered after treatment. The effects of cannabinoids on invasiveness could be blocked by the application of receptor antagonists and are likely mediated via CB1/CB2. In conclusion, our results suggest that cannabinoids can influence glioblastoma cell invasion in a receptor and cell type specific manner that is independent of proliferation and apoptosis. Thus, cannabinoids can potentially be used in the future as an addition to current therapy.

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