Cannabidiol, A Non-Psychoactive Cannabinoid Compound, Inhibits Proliferation And Invasion In U87-Mg And T98G Glioma Cells Through A Multitarget Effect

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http://www.ncbi.nlm.nih.gov/pubmed/24204703

PLoS One. 2013 Oct 21;8(10):e76918. doi: 10.1371/journal.pone.0076918.

Cannabidiol, a Non-Psychoactive Cannabinoid Compound, Inhibits Proliferation and Invasion in U87-MG and T98G Glioma Cells through a Multitarget Effect.

Solinas M, Massi P, Cinquina V, Valenti M, Bolognini D, Gariboldi M, Monti E, Rubino T, Parolaro D.

Source

Department of Theoretical and Applied Sciences, Biomedical Research Division, Centre of Neuroscience, University of Insubria, Busto Arsizio, Varese, Italy.

Abstract

In the present study, we found that CBD inhibited U87-MG and T98G cell proliferation and invasiveness in vitro and caused a decrease in the expression of a set of proteins specifically involved in growth, invasion and angiogenesis. In addition, CBD treatment caused a dose-related down-regulation of ERK and Akt prosurvival signaling pathways in U87-MG and T98G cells and decreased hypoxia inducible factor HIF-1α expression in U87-MG cells. Taken together, these results provide new insights into the antitumor action of CBD, showing that this cannabinoid affects multiple tumoral features and molecular pathways. As CBD is a non-psychoactive phytocannabinoid that appears to be devoid of side effects, our results support its exploitation as an effective anti-cancer drug in the management of gliomas.

Effect of increasing CBD concentrations on U87-MG (A) and T98G © glioma cell invasion.

U87-MG and T98G cells were treated with CBD, seeded onto a filter coated with matrigel and incubated for 24 h at 37°C. Cells that invaded the lower surface of the filter were quantified. The invasion was expressed as percentage of the untreated control. Data represent the mean ± S.E.M. of three independent experiments. **p<0.01 vs Control [C], Dunnett’s t test. Effect of increasing CBD concentrations on U87-MG ( and T98G (D) glioma cell viability. U87-MG and T98G glioma cells were cultured in serum-free medium with increasing concentrations of CBD. Cell viability was determined by MTT assay after 24 h of treatment. The viability was expressed as percentage of the untreated control. Data represent the mean ± S.E.M. of at least three independent experiments.

Effect of increasing CBD concentrations on the protein expression profile of U87-MG glioma cells.

U87-MG cells were treated with CBD for 24 h and supernatants were used to determine different protein levels through a human array kit/proteome profiler. (A) Representative proteomic membrane analysis with indication of the modified proteins. ( Densitometric analysis of the membrane spots reported as percentage of the untreated control. Data represent the mean ± S.E.M. of three independent experiments. **p<0.01, ***p<0.001 vs Control, Dunnett’s t test. U87-MG cells were treated with CBD for 24 h and lysates from cells were used to assess MMP-9 and TIMP-4 protein levels. © Western blot analysis of MMP-9 and TIMP-4. A representative western blot is shown. (D) Densitometric analysis of MMP-9 and TIMP-4 signal bands from three independent experiments is reported. **p<0.01 vs Control [C], Dunnett’s t test.

Effect of increasing CBD concentrations on the protein expression profile of T98G glioma cells.

T98G cells were treated with CBD for 24/proteome profiler. (A) Representative proteomic membrane analysis with indication of the modified proteins. ( Densitometric analysis of the membrane spots reported as percentage of the untreated control. Data represent the mean ± S.E.M. of three independent experiments. **p<0.01 vs Control, Dunnett’s t test.

Effect of increasing CBD concentrations on ERK phosphorylation in U87-MG and T98G glioma cells.

U87-MG and T98G cells were treated with CBD for 24 h and lysates from cells were used to assess ERK protein levels. (A,C) Western blot analysis of phospho- and total-ERK. A representative western blot is shown. (B,D) Densitometric analysis of ERK signal bands from three independent experiments is reported. *p<0.05, **p<0.01 vs Control [C], Dunnett’s t test.

Effect of increasing CBD concentrations on Akt phosphorylation in U87-MG and T98G glioma cells.

U87-MG and T98G cells were treated with CBD for 24 h and lysates from cells were used to assess Akt protein levels. (A,C) Western blot analysis of phospho- and pan-Akt. A representative western blot is shown. (B,D) Densitometric analysis of Akt signal bands from three independent experiments is reported. *p<0.05, **p<0.01 vs Control [C], Dunnett’s t test.

Effect of CBD on HIF-1α level in glioma cells grown in normoxic and hypoxic conditions.

U87-MG and T98G glioma cells were grown under normoxia and two different hypoxic conditions (see Materials and methods). Cells were treated with CBD for 24 h and lysates from the cells were used to assess HIF-1α protein levels. (A–C) Western blot analysis of HIF-1α and β-actin. A representative western blot is shown. (B–D) Densitometric analysis of HIF-1α signal bands from three independent experiments is reported. *p<0.05, **p<0.01 vs Control [C], ° p<0.05, °° p<0.01, °°° p<0.001, °°°° p<0.0001 vs normoxia Control [C], Dunnett’s t test.

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Resumindo: maconha cura o câncer.