| Objective: Esophageal carcinoma is one of the high incidence indigestive malignant tumor. At present, incidence and mortality is increasingyear by year. It is popular in China,where the incidence of it is the first in theworld. At present the main treatment is including surgery, chemotherapy,radiation therapy, and biological treatment. Most patients has found thatbelongs to middle-late stage because of the lack of specificity on earlystage.Therefore, it is urgently needed that exploring more effective methodsand treatments to improve the patient's prognosis and quality of life. In recentyears, many reports show that dihydroartemisinin can induce anti-neoplasticactivity on many human tumor cell lines. At present, rare has been elucidatedon the influence of esophagealcancer cells by inhibitor of dihydroartemisinin.This experiment is aimed to investigate the effect of dihydroartemisinin onproliferation,apoptosis and expression of associated protein of humanesophageal carcinoma cell line TE-13in vitro in order to further investigatepossible mechanism of dihydroartemisinin inhibiting esophageal carcinomacell line. This will provide the theoritic basis for esophageal carcinomatherapy and chemical prevention.Methods:1Esophageal carcinoma cells were incubated in culture medium in vitro.Using microscope observed the change of morphologic of TE-13cells whichwere treated with different concentrations of DHA.2Human esophageal cancer line TE-13were incubated with differentconcentration of DHA in vitro.MTT assay were used to determine theinfluence of DHA on proliferation of esophageal cancer line TE-13.3The influence of DHA on apoposis of human esophageal cancer lineTE-13was determine by flow cytomery. 4The influence of DHA on cell cycle of human esophageal cancer lineTE-13was determine by flow cytomery.5The expression of Survivin, Caspase-3protein was qualitationly studiedby immunocytochemistry staining after treated with DHA.Results:1The cells untreated with DHA were grow adherently,distributed,diamond or polygonal,and they were diaphanous,well-stacked andin a good condition. But the cells treated with DHA were shrinked and broken,and became irregular in shape,the number of cells,reducedshrinkage,cytoplasmic vacuoles,nuclear condensation,off the apparent increasein the number of cells,cell bodies of refraction weakened.2DHA on TE-13cell growth inhibition: MTT colorimetric methodshowed that DHA (10,20,40,80,160μmol/l)could inhibit the proliferation ofTE-13cells. After treated with DHA for24h to72h, compared with controlgroup, the OD values of DHA-treated groups decreased, and there wasstatistically significant difference between control group and every treatmentgroup (p<0.05). Moreover, with the increasing concentration of DHA andprolonging of treatment time, the speed of cell proliferation decreasedgradually, the inhibitory rate increased, in other words, DHA inhibited theproliferation of TE-13cells significantly in a dose-dependent andtime-dependent manner.3DHA on analysis of TE-3: When cells were harvested for the analysison apoptosis by flow cytometry, the results were: DHAcan induce apoptosis inhuman colonic cancer cell line TE-13.After TE-13cells treated withDHA(40,80,160μmol/L) for48h,the apoptotic percentage were8.643%±0.126%,18.606%±0.776%,29.026%±0.730%. With the increasingconcentrations of DHA,the apoptosis rate were enhanced,and there differencehad statistical siginificance compared the control group (P<0.01).4DHA on cell cycle of TE-13: DHA could induce an arrest of cell cyclein G0/G1in a dose-dependent manner. After TE-13cells were treated with40,80,160μmol/L DHA for48h, with the increasing concentration of DHA, the number of cells in G0/G1phase increased gradually, while the number of cellsin S and G2/M phase decreased grudually. With the increasing concentrationsof DHA, the number of cells in G0/G1phase were increased,and theredifference had statistical siginificance compared the control group (P<0.05).5The expressiong of Survivin and Caspase-3protein of TE-13cellsanalyzed by immunocytochemical method.After scoring and statistics,expressive values of Survivin (IHSvalue)decreased with the increasingconcentration of DHA. Expressive values of Caspase-3(IHS value) increasedwith the increasing concentration of DHA.To the expressive values of bothproteins, there was statistically significant difference between the negativecontrol group and every DHA-treated group(p <0.01).Conclusions:1DHA has the effect of anti-varian cancer cell line TE-13in vitro in adose and time-dependent manner.2DHA can induce apoptosis and arrest cell cycle at G0/G1phase inhuman colonic cancer cell line TE-13,it has a dose-dependent andtime-dependent relationship.3Within a certain drug concentration, TE-13can increase the expressionof Caspase-3and decrease the expression of Survivin in TE-13cells. We inferthat DHA's effects of inhibiting proliferation and inducing apoptosis in TE-13cells seem to due to up-regulation of the expression of Caspase-3protein anddown-regulation of the expression of Survivin. |
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