Journal: Biomedicines

Article Title: The Sesquiterpene Lactone Cynaropicrin Manifests Strong Cytotoxicity in Glioblastoma Cells U-87 MG by Induction of Oxidative Stress

doi: 10.3390/biomedicines10071583

Figure Lengend Snippet: Cytotoxic effects of Cyn on human glioblastoma U-87 MG cells. ( A ) Molecular structure of SL Cynaropicrin (IUPAC name: ([(3 a R,4S,6 a R,8S,9 a R,9 b R)-8-hydroxy-3,6,9-trimethylidene-2-oxo-3 a ,4,5,6 a ,7,8,9 a ,9 b -octahydroazuleno[4,5- b ]furan-4-yl] 2-(hydroxymethyl)prop-2-enoate). ( B ) Determination of IC50 values on U-87 MG cells using GraphPad Prism 7 software after 24, 48 and 72 h of incubation with Cyn 0.01, 0.1, 1, 10, 25, 50 and 100 µM and DMSO 0.1% as the vehicle control. ( C ) Dose and time-dependent reduction of the U-87 MG cell number under daily exposure to Cyn 4, 8 and 10 µM for 24, 48 and 72 h. ( D ) Morphological change of 4, 8 and 10 µM Cyn-treated U-87 MG cells for 24, 48 and 72 h. White arrows indicate round-shaped cells with a loss of filaments and the presence of cell shrinkage. ( E ) Cytotoxic effects of daily exposure to Cyn 4, 8 and 10 µM for 24, 48 and 72 h on the U-87 MG cell metabolism. ( F ) Effect of Cyn 4 µM on the U-87 MG clonogenic potential. For all the experiments, values are the mean ± SEM of 3 individual determinations. One-way ANOVA test, p -value < 0.05. According to GraphPad Prism 7 software, **** p -value < 0.0001 (extremely significant).

Article Snippet: Human continuous glioblastoma cell line U-87 MG was purchased from the Sigma Aldrich Collection (LGC Promochem, Teddington, UK).

Techniques: Software, Incubation, Control

Journal: Biomedicines

Article Title: The Sesquiterpene Lactone Cynaropicrin Manifests Strong Cytotoxicity in Glioblastoma Cells U-87 MG by Induction of Oxidative Stress

doi: 10.3390/biomedicines10071583

Figure Lengend Snippet: Cyn-induced oxidative stress in human glioblastoma U-87 MG cells. ( A ) Pretreatment of U-87 MG cells with NAC 3 mM for 4 h, followed by incubation for 24 h with Cyn 0.01, 0.1, 1, 10, 25, 50 and 100 μM. DMSO 0.1% was used as the vehicle control. ( B ) Preincubation with NAC 3 mM for 4 h preserved U-87 MG from Cyn-induced morphological change, since the cells maintained their protrusions rather than appeared round-shaped (white arrows). ( C ) Quantitative analysis of ROS generation in U-87 MG cells under 2 and 6 h of treatment with Cyn 8 and 25 μM. DMSO 0.1% was used as the vehicle control. ( D ) Qualitative analysis of ROS production after 2 h of exposure to Cyn 8 and 25 μM. ( E ) Immunofluorescence staining of NRF2 in U-87 MG treated for 24 h with Cyn 25 µM, which showed a nuclear localization with respect to the control cells treated with 0.1% DMSO (white arrows). Magnification 20×. Data were analyzed by a Student’s t -test, p -value < 0.05. According to GraphPad Prism 7 software, * p -values from 0.01 to 0.05 (significant) and ** p -values from 0.001 to 0.01 (very significant).

Article Snippet: Human continuous glioblastoma cell line U-87 MG was purchased from the Sigma Aldrich Collection (LGC Promochem, Teddington, UK).

Techniques: Incubation, Control, Immunofluorescence, Staining, Software

Journal: Biomedicines

Article Title: The Sesquiterpene Lactone Cynaropicrin Manifests Strong Cytotoxicity in Glioblastoma Cells U-87 MG by Induction of Oxidative Stress

doi: 10.3390/biomedicines10071583

Figure Lengend Snippet: Cytotoxic effects of Cyn on patient-derived glioblastoma cell lines NULU and ZAR. ( A ) Cytotoxic effects of daily exposure to Cyn 4, 8 and 10 µM for 24, 48 and 72 h on NULU ( A ) and ZAR ( B ) cell metabolism. DMSO 0.1% was used as the vehicle control. ( C ) Morphological changes for the 4, 8 and 10 µM Cyn-treated NULU and ZAR cell lines for 24, 48 and 72 h. Data were analyzed by the Student’s t -test, p -value < 0.05. According to GraphPad Prism 7 software, * p -value 0.01–0.05 was considered statistically significant, ** p -value 0.001–0.01 very significant, *** p -value 0.0001–0.001 extremely significant and while **** p -values < 0.0001 were extremely significant. Taking together, these results clearly indicate the potentialities of Cyn as adjuvant therapy to conventional chemotherapy TMZ in IDH-mutant and wild-type glioblastoma.

Article Snippet: Human continuous glioblastoma cell line U-87 MG was purchased from the Sigma Aldrich Collection (LGC Promochem, Teddington, UK).

Techniques: Derivative Assay, Control, Software, Adjuvant, Mutagenesis

Journal: bioRxiv

Article Title: Optimizing Angiopep-2 Density on Polymeric Nanoparticles for Enhanced Blood-Brain Barrier Penetration and Glioblastoma Targeting: Insights from In Vitro and In Vivo Experiments

doi: 10.1101/2024.11.05.622195

Figure Lengend Snippet: In vitro drug release of Dox from (A) Dox@DAA 100 and (B) Dox@DAA 100 -Ang-2(1:2) at pH 5 and pH 7.4, using the two-way ANOVA with Tukey’s multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. (C) Cytotoxicity assessment of various concentrations of Free Dox, Dox@DAA 100 and Dox@DAA 100 -Ang-2(1:2) (0.01, 0.05, 0.10, and 0.50 µM) on U87 after 72 h treatment, using the two-way ANOVA with Tukey’s multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. All data are shown as mean ± SD, n = 3. (D) Schematic of the BBB-GBM-on-a-chip model consisting of two channels. The blood channel was formed by a monolayer of hCMEC/D3 cells over 3 µm microchannels under a fluidic environment. The brain channel was seeded by U87 cells receiving medium and Dox-loaded nanoparticles from the blood channel. Relative apoptosis levels (n-fold change to the untreated control chips) of activated caspase-3/7 (E) in “blood” channels and (F) in “brain” channels, respectively, using the one-way ANOVA with Tukey’s multiple comparisons test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, n = 3. All data are shown as mean ± SD.

Article Snippet: Immortalized human brain microvascular endothelial cells (hCMEC/D3) cells and human glioblastoma U87 cells were purchased from Merck.

Techniques: In Vitro, Control

Journal: Nature Communications

Article Title: Chemogenetic ON and OFF switches for RNA virus replication

doi: 10.1038/s41467-021-21630-5

Figure Lengend Snippet: a Subcutaneous human glioma U87 xenografts were intratumorally treated with luciferase-expressing VSV-Pprot-Luc with (+PI) or without (-PI) concomitant intraperitoneal (i.p.) application of APV. Representative bioluminescence (BLI) images are shown from 8 days after virus injection. b BLI quantification of luciferase signal from VSV-Pprot-Luc treated tumors in mice receiving PI cocktail (APV + RTV) (red) or drug vehicle (blue) (technical replicates; n = 5; * denotes significantly different measurements with p < 0.05; unpaired two-tailed t test at indicated time points). c , d Single intratumoral treatment of subcutaneus U87 tumors with VSV ( n = 6) or VSV-Lprot-GFP with ( n = 5) or without ( n = 5) PI treatment (APV + RTV) (mock n = 6). Log-rank (Mantel-Cox) test was performed (** p = 0.0052). e – h 2 μl of VSV-DsRed or VSV-Pprot-GFP were stereotactically injected into the striatum of BALB/c mice. PI (APV + RTV) treatment was applied every 12 h for 9 days. VSV-Pprot-GFP was well tolerated with no significant weight loss ( e ) or signs of neurotoxicity ( f ) compared to fatal neurotoxicity of parental VSV-DsRed ( g ) (** p = 0.0078; Log-rank (Mantel-Cox) test). Symbols in f display score per mouse per time point). h Histological fluorescence analysis of coronal brain sections revealed extended spread of VSV-DsRed in the striatum, subcortical areas and hypothalamus (bilateral) at 3 days post inoculation (dpi). In contrast, GFP expression from VSV-Pprot-GFP (10 dpi) was restricted to the immediate lining of the injection needle track without any signs of intracranial spread, irrespective of i.p. co-treatment of PI or drug vehicle. PI treatment with high-dose IDV to increase CNS availability after VSV-Lprot-GFP injection did also not induce signs of neurotoxicity and brain parenchymal spread was restricted to the injection site ( i , j ); n = 3 for VSV-GFP, n = 5 for Lprot variants. Bioluminescence experiments in panels ( a , b ) and associated tumor growth and survival study ( c , d ) were performed once. Intracranial injection experiments ( e – h ) were performed two times. Source data are provided in the Source Data File.

Article Snippet: For subcutaneous xenografts, 100 μl glioblastoma cell suspension containing 2 × 10 6 human U87 or G62 glioma cells were injected into the right flanks of NMRI nude mice (Janvier) or NOD-SCID mice (Janvier), respectively.

Techniques: Luciferase, Expressing, Virus, Injection, Two Tailed Test, Fluorescence

Journal: Frontiers in Molecular Biosciences

Article Title: Proteomic Analysis on Anti-Proliferative and Apoptosis Effects of Curcumin Analog, 1,5-bis(4-Hydroxy-3-Methyoxyphenyl)-1,4-Pentadiene-3-One-Treated Human Glioblastoma and Neuroblastoma Cells

doi: 10.3389/fmolb.2021.645856

Figure Lengend Snippet: The cell viability in percentage (%) of (A) U-87 MG glioblastoma and (B) SH-SY5Y neuroblastoma cells upon treatment with MS13 and curcumin. The concentration (ranged from 0 to 100 μM) of MS13 or curcumin was log10 transformed. The cell viability of both cells decreased as the concentration of MS13 or curcumin increased. Results are expressed as the average of percentage of cell viability. Error bars represent mean ± standard error mean. The experiments were performed in triplicates and results were compared between three biological replicates. Statistical analysis was performed using analysis of variance (ANOVA), comparing cell viability of each concentration with the untreated sample. Asterisks indicate the difference in the statistical significance of the mean values between treated and untreated cells. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human glioblastoma (U-87 MG) cell line was purchased from ElabScience ® (Wuhan, China), whereas human neuroblastoma (SH-SY5Y) cell line and normal human epithelial hepatocytes, WRL-68 were obtained from the American Type Culture Collection (ATCC, Rockville, MD, United States).

Techniques: Concentration Assay, Transformation Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Proteomic Analysis on Anti-Proliferative and Apoptosis Effects of Curcumin Analog, 1,5-bis(4-Hydroxy-3-Methyoxyphenyl)-1,4-Pentadiene-3-One-Treated Human Glioblastoma and Neuroblastoma Cells

doi: 10.3389/fmolb.2021.645856

Figure Lengend Snippet: The anti-proliferative effect of MS13 and curcumin on (A) U-87 MG glioblastoma and (B) SH-SY5Y neuroblastoma cells at 24, 48, and 72 h. Results are expressed as the average of cell viability in percentage (%) against concentration (μM). Error bars represent mean ± standard error mean. All experiments were performed in triplicates and results were compared between three biological replicates. Statistical analysis was performed using ANOVA, comparing cell viability of each concentration with the untreated sample. Asterisks indicate the difference in the statistical significance of the mean values between treated and untreated cells. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human glioblastoma (U-87 MG) cell line was purchased from ElabScience ® (Wuhan, China), whereas human neuroblastoma (SH-SY5Y) cell line and normal human epithelial hepatocytes, WRL-68 were obtained from the American Type Culture Collection (ATCC, Rockville, MD, United States).

Techniques: Concentration Assay