Antiproliferative Effect Of Extracts From Kazakh Medicinal Herb Daphne Altaica Pall On Human Esophageal Carcinoma ECa-109 Cells

Objective: to evaluate whether ethyl acetate extract,n-Hexane extract and methanol extract obtained from Daphne altaica Pall has any specific antiproliferative effect on human esophageal carcinoma ECa-109 cells;to study the mechanism of its antiproliferative activity at cell cycle arrest,apoptosis and PPARγ gene expression level;to provide some basic evidence for the further research.Methods:The growth inhibitory effect of three extracts of D.altaica on human esophageal carcinoma ECa-109 cells was evaluated by MTT assay.Cell apoptosis was measured by flow cy tometry after Annexin single-staining.Cell cycle was measured by flow cytometry after PI.Changes in cellular morphology was observed using inverted fluorescence microscopy.The level of PPARγ mRNA and protein expression in Eca-109 cells was detected using Westernblot and Q-PCR..Results:Da-Ea,Da-Hx,Da-Me extracts exhibited a significant antiproliferative effect on human esophageal carcinoma ECa-109 cells in both time and concentration dependent manners,Compared with the control group,the inhibition effect was significant(P<0.05);After treated by Daphne altaica Pall.extract at different time points Eca-109 cells appeared to be morphologically changed:,polygonal-shaped cells became elongated,the nucleus was broken and the number of dead cells was increased in time and concentration-dependent manners.Cell integrity was damaged more seriously by the time increase;DaEa,Da-Hx,Da-Me extracts induced apoptosis and S phase arrest in esophageal carcinoma ECa-109 cells.Compared with the control group,the apoptosis level at 96 h were significantly increased(P<0.05),the number of cells in S phase was significantly increased(P<0.01),whereas number of cells in G1/G2 phase was dramatically reduced(P<0.01).Expression level of PPARγ mRNA in esophageal carcinoma Eca-109 cells was increased remarkably after treated by Da-Ea,Da-Hx,Da-Me: Da-Ea can increase the expression of PPARγ mRNA in the esophageal carcinoma Eca-109 cells at 24 h,48 h,72 h(P<0.05),while Da-Hx,Da-Me can increase it’s expression at 48 h,72 h(P<0.05).Expression level of PPARγ protein increased as well(P<0.05).Conclusion:This study confirmed that the Daphne altaica Pall.extracts had antiproliferative effect in vitro,Its antiproliferative activity was associated with an induction of S phase arrest and cell apoptosis as well as high expression level of PPARγ gene in ECa-109 cells.