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Cellular & Molecular Biology Letters logoLink to Cellular & Molecular Biology Letters
. 2014 Jul 7;19(3):347–360. doi: 10.2478/s11658-014-0200-x

Cannabinoid receptor activation inhibits cell cycle progression by modulating 14-3-3β

Hye-Won Jung 1, Inae Park 1, Sungho Ghil 1,✉
PMCID: PMC6275927  PMID: 25002257

Abstract

Cannabinoids display various pharmacological activities, including tumor regression, anti-inflammatory and neuroprotective effects. To investigate the molecular mechanisms underlying the pharmacological effects of cannabinoids, we used a yeast two-hybrid system to screen a mouse brain cDNA library for proteins interacting with type 1 cannabinoid receptor (CB1R). Using the intracellular loop 3 of CB1R as bait, we identified 14-3-3β as an interacting partner of CB1R and confirmed their interaction using affinity-binding assays. 14-3-3β has been reported to induce a cell cycle delay at the G2/M phase. We tested the effects of cannabinoids on cell cycle progression in HeLa cells synchronized using a double-thymidine block-and-release protocol and found an increase in the population of G2/M phase cells. We further found that CB1R activation augmented the interaction of 14-3-3β with Wee1 and Cdc25B, and promoted phosphorylation of Cdc2 at Tyr-15. These results suggest that cannabinoids induce cell cycle delay at the G2/M phase by activating 14-3-3β.

Keywords: Cannabinoids, Cdc2, Cdc25B, Cyclin B, G2/M phase, GIPs, HeLa, Phosphorylation, Wee1, Yeast-two hybrid

Full Text

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Abbreviations used

CBR

cannabinoid receptors

CB1R

type 1 CBR

CB2R

type 2 CBR

GIPs

GPCR-interacting proteins

GPCR

G protein-coupled receptor

GPR55

G protein-coupled receptor 55

GST-14-3-3β

glutathione-S-transferase-tagged 14-3-3β

HA-CB1R

3xHA-tagged CB1R

IL3

intracellular loop 3 domain

PBS

phosphate-buffered saline

PI

propidium iodide

PPAR-γ

peroxisome proliferator-activated receptor-γ

SDS

sodium dodecyl sulfate

Footnotes

These authors contributed equally to this work

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