DHA induced cytotoxicity in NSCLC cells via inhibiting PI3K pathway activation and GLUT2 expression
10.3969/j.issn.1000-4718.2015.11.007
- VernacularTitle:DHA通过抑制PI3K通路和GLUT2表达而诱导NSCLC细胞毒性
- Author:
Fasheng MIN
;
Jianhong YANG
;
Jianjun HUANG
;
Qiliang LIU
- Publication Type:Journal Article
- Keywords:
Non-small cell lung cancer;
Dihydroartemisinin;
PI3K pathway;
Glucose transporter 2;
Cytotox-icity
- From:Chinese Journal of Pathophysiology
2015;(11):1961-1969
- CountryChina
- Language:Chinese
-
Abstract:
[ ABSTRACT ] AIM: To investigate the molecular mechanisms of cytotoxicity induced by dihydroartemisinin ( DHA) in non-small cell lung cancer ( NSCLC) cells.METHODS:NSCLC cell lines A549 and NCI-H1650 were treated with various concentrations of DHA for indicated time.Subsequently, the effects of DHA on the cell activity, colony forma-tion ability and apoptosis were determined by MTT assay, colony formation assay, Annexin V-FITC/PI staining and flow cy-tometry, respectively.At the same time, the effects of DHA on glucose, ATP and lactate levels were assessed, and the PI3K pathway activation and glucose transporter 2 ( GLUT2) expression were detected by Western blot in the A549 cells and NCI-H1650 cells.Overexpression of GLUT2 and Rheb was established in A549 and NCI-H1650 cells by transfection with GST-GLUT2 and GST-Rheb plasmids, respectively, and the effects of DHA on cell activity, apoptosis, glucose level, ATP content and PI3K pathway activation were analyzed in A549 cells and NCI-H1650 cells.The effect of glucose depriva-tion on the cytotoxicity triggered by DHA in NSCLC cells was also determined.RESULTS:Compared with control group, DHA significantly inhibited cell activity and colony formation ability, and induced remarkable cell apoptosis in the A549 cells and NCI-H1650 cells.At the same time, DHA reduced ATP and lactate contents, and hindered glucose uptake in a time-and dose-dependent manner in A549 cells and NCI-H1650 cells.The activity of PI3K pathway and GLUT2 expression were downregulated, while upregulated GLUT2 expression and activated PI3K pathway reduced the cytotoxicity induced by DHA in NSCLC cells.Glucose deprivation increased DHA-mediated cytotoxicity in NSCLC cells.On the contrary, high levels of glucose inhibited DHA-mediated cytotoxicity in NSCLC cells.CONCLUSION: DHA restrains cell activity and colony formation, and induces apoptosis.DHA induces cytotoxicity via inhibiting PI3K pathway activation and GLUT2 ex-pression, leading to inhibit glycolytic metabolism in NSCLC cells.
