Cannabidiol Regulates Biological Behaviors of Endometrial Cancer Cells Through PERK-eIF2α-ATF4 Signaling Pathway
[Objective]To investigate the effect and mechanism of cannabidiol(CBD)on biological behaviors of endometrial cancer cells.[Methods]Endometrial cancer Ishikawa and KLE cells were divided into control group,dimethyl sulfoxide(DMSO)group,CBD+DMSO group,and CBD+DMSO+GSK2606414[a protein kinase RNA-like endoplasmic reticulum kinase(PERK)inhibitor]group.The effects of CBD on prolifera-tion,migration,invasion and apoptosis of endometrial cancer cells were evaluated by CCK-8 assay,scratch healing assay,Transwell assay and flow cytometry,respectively.The level of reactive oxygen species(ROS)in cells was measured with ROS probes.The expression of CCAAT-enhancer-binding protein homologous protein(CHOP)and PERK-eukaryotic translation initiation factor 2α-activating transcription factor 4(eIF2α-ATF4)signaling pathway-related proteins in endometrial cancer cells was detected with Western blot.The PERK signaling pathway inhibitor GSK2606414 was used to validate the pathway.[Results]Cannabidiol ex-hibited concentration-and time-dependent inhibition of proliferation of Ishikawa and KLE cells.Cannabidiol significantly inhibited Ishikawa cell proliferation at concentrations ranging from 2 to 16 p.M(P＜0.05)and KLE cell proliferation at concentrations ranging from 6 to 16 pM(P＜0.05).After Cannabidiol intervention for ≥24 h,there was a significant inhibition of proliferation in both Ishikawa and KLE cell(P＜0.05).Cannabidiol also inhibited the migration,invasion,and induced apoptosis of Ishikawa and KLE cell(P all＜0.05).The 24 h healing rate of Ishikawa cell in the CBD+DMSO group was 37.54％±0.97％,and the 48 h healing rate was 47.87％±1.46％,for KLE cells,the 24 h healing rate was 41.93％±2.85％,and the 48h healing rate was 51.29％±0.75％.Cannabidiol increased intracellular ROS levels(Ishikawa P=0.007 4,KLE P＜0.001).Following Cannabidiol treatment in Ishikawa and KLE cell,the expression levels of CHOP,phosphorylated PERK(p-PERK),phosphorylated elF2α(p-eIF2α),and ATF4 were significantly increased compared to the control and DMSO groups(P＜0.001).After the addition of GSK2606414,the expression levels of p-PERK(P＜0.001),p-eIF2α(P＜0.001),and ATF4(P＜0.001)were downregulated compared to the CBD+DMSO group but was higher than the control and DMSO groups.[Conclusion]In endometrial cancer cells,cannabidiol inhibits cell proliferation,migration,and invasion capabilities,induces apoptosis,and thus exerts anti-tumor effects by activating the PERK-eIF2α-ATF4 signaling pathway.Its anti-tumor effects may be associated with the ac-tivation of endoplasmic reticulum stress and the accumulation of ROS.
cannabidiolendometrial cancerendoplasmic reticulum stressreactive oxygen species
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