RESUMO
Daphne altaica Pall. (D. altaica; Thymelaeaceae) has long been used in traditional Kazakh medicine for the treatment of cancer and respiratory diseases. Previous studies have demonstrated the in vitro anticancer effects of D. altaica extract and its constituents in certain cancer cell lines; however, the underlying molecular mechanisms are not completely understooD. The present study aimed to investigate the molecular mechanisms underlying the activity of an ethyl acetate extract of D. altaica (DaEa) by assessing its effects on cell morphology, cell apoptosis, cell cycle progression and the expression levels of peroxisome proliferatoractivated receptor γ (PPARγ) in Eca109 cells. Cell morphology was observed under a phase contrast microscope. Cell apoptosis and cell cycle progression were assessed by flow cytometry following Annexin V/propidium iodide (PI) double staining and PI single staining, respectively. The mRNA and protein expression levels of PPARγ were determined by reverse transcriptionquantitative PCR and western blotting, respectively. Compared with the control group, the percentage of apoptotic cells, cell cycle arrest at S phase and apoptotic morphological cell characteristics were increased in DaEatreated Eca109 cells. Furthermore, DaEa treatment upregulated the mRNA and protein expression levels of PPARγ compared with the control cells. Highperformance liquid chromatography with diodearray detection indicated that daphnetin7OßDglucoside, daphnetin, demethyldaphnoretin7OßDglucopyranoside and genkwanol A were the main constituents of DaEa. Collectively, the results suggested that DaEa displayed antiproliferative activities in Eca109 cells by inducing apoptosis and S phase cell cycle arrest, as well as upregulating PPARγ expression levels.