Over-expressed human TREK-1 inhibits CHO cell proliferation via inhibiting PKA and p38 MAPK pathways and subsequently inducing G1 arrest  

作　　者：Man ZHANG Hua-jing YIN Wei-ping WANG Jiang LI Xiao-liang WANG 

机构地区：[1]State Key Laboratory of Bioactive Substances and Functions of Natural Medicines, Institute of Materia Medica, Chinese Academy ofMedical Sciences and Peking Union Medical College, Beijing 100050, China

出　　处：《Acta Pharmacologica Sinica》2016年第9期1190-1198,共9页中国药理学报（英文版）

摘　　要：Aim： Recent studies have shown that the two-pore-domain potassium channel TREK-1 is involved in the proliferation of neural stem cells, astrocytes and human osteoblasts. In this study, we investigated how TREK-1 affected the proliferation of Chinese hamster ovary （CHO） cells in vitro. Methods： A CHO cell line stably expressing hTREK-1 （CHO/hTREK-1 cells） was generated. TREK-1 channel currents in the cells were recorded using whole-cell voltage-clamp recording. The cell cycle distribution was assessed using flow cytometry analysis. The expression of major signaling proteins involved was detected with Western blotting. Results： CHO/hTREK-1 cells had a high level of TREK-1 expression, reached up to 320%±16% compared to the control cells. Application of arachidonic acid （10 μmol/L）, chloroform （1 mmol/L） or etomidate （10 μmol/L） substantially increased TREK-1 channel currents in CHO/hTREK-1 cells. Overexpression of TREK-1 caused CliO cells arresting at the G1 phase, and significantly decreased the expression of cyclin DI. The TREK-1 inhibitor/-butylphthalide （1-100 μmol/L） dose-dependently attenuated TREK-l-induced G1 phase cell arrest. Moreover, overexpression of TREK-1 significantly decreased the phosphorylation of Akt （S473）, glycogen synthase kinase-3β （S9） and cAMP response element-binding protein （CREB, S133）, enhanced the phosphorylation of p38 （T180/Y182）, but did not alter the phosphorylation and expression of signal transducer and activator of transcription 3 （STAT3）. Conclusion： TREK-1 overexpression suppresses CliO cell proliferation by inhibiting the activity of PKA and p38/MAPK signaling pathways and subsequently inducing G1 phase cell arrest.Aim： Recent studies have shown that the two-pore-domain potassium channel TREK-1 is involved in the proliferation of neural stem cells, astrocytes and human osteoblasts. In this study, we investigated how TREK-1 affected the proliferation of Chinese hamster ovary （CHO） cells in vitro. Methods： A CHO cell line stably expressing hTREK-1 （CHO/hTREK-1 cells） was generated. TREK-1 channel currents in the cells were recorded using whole-cell voltage-clamp recording. The cell cycle distribution was assessed using flow cytometry analysis. The expression of major signaling proteins involved was detected with Western blotting. Results： CHO/hTREK-1 cells had a high level of TREK-1 expression, reached up to 320%±16% compared to the control cells. Application of arachidonic acid （10 μmol/L）, chloroform （1 mmol/L） or etomidate （10 μmol/L） substantially increased TREK-1 channel currents in CHO/hTREK-1 cells. Overexpression of TREK-1 caused CliO cells arresting at the G1 phase, and significantly decreased the expression of cyclin DI. The TREK-1 inhibitor/-butylphthalide （1-100 μmol/L） dose-dependently attenuated TREK-l-induced G1 phase cell arrest. Moreover, overexpression of TREK-1 significantly decreased the phosphorylation of Akt （S473）, glycogen synthase kinase-3β （S9） and cAMP response element-binding protein （CREB, S133）, enhanced the phosphorylation of p38 （T180/Y182）, but did not alter the phosphorylation and expression of signal transducer and activator of transcription 3 （STAT3）. Conclusion： TREK-1 overexpression suppresses CliO cell proliferation by inhibiting the activity of PKA and p38/MAPK signaling pathways and subsequently inducing G1 phase cell arrest.

关 键 词：TREK-l CliO cells G1 phase arrest cell proliferation /-butylphthalide PKA p38/MAPK 

分 类 号：R[医药卫生]