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