摘要
目的观察青蒿琥酯(artesunate,Art)对人食管癌Eca-109细胞系的抑瘤作用,并进一步探讨Art诱导肿瘤细胞周期阻滞与CDC25A、TGF-β表达的关系。方法体外培养人食管癌Eca-109细胞系及正常人外周血单个核细胞(hPBMC),利用MTT法测定细胞增殖;采用流式细胞术(FCM)测定细胞周期;应用RT-PCR方法检测CDC25AmRNA表达,应用Western blot方法检测蛋白表达。结果Art能显著抑制Eca-109细胞的增殖,IC50为(68.80±0.76)μmol/L,而对hPBMC的增殖则没有明显抑制作用。低浓度Art可将细胞阻滞于G0/G1期,S期细胞显著减少,当浓度达到100μmol/L时,细胞阻滞于G2/M期。Art可显著抑制Eca-109细胞CDC25AmRNA及蛋白表达,同时显著上调TGF-β的蛋白表达水平。结论Art可抑制肿瘤细胞生长,上调TGF-β表达,抑制CDC25A表达。
Objective To observe the effects of Art on the proliferation of human esophageal carcinoma Eca-109 cell line and to explore the relationship between cell cycle arrest effect of Art and the expression of CDC25A and TGF-β. Methods Human esophageal carcinoma Eca-109 cells and healthy human peripheral blood mononuclear cells （hPBMCs） were cultured. The inhibitory effects of Art on cell proliferation were determined by MTT method. The changes of cell cycle of tumor cells were assayed by flow cytometry （FCM）. The expression of CDC25A mRNA was examined by RT-PCR and the protein expression of CDC25A as well as TGF-β were determined by Western blot. Results Art could significantly inhibit the proliferation of Eca-109 cells. Its IC50 was （68.80 ± 0. 76 ） μmol/L. It had weaker effect on the proliferation of hPBMC. The cell cycle of Eca-109 was greatly changed after the administration of Art. The number of Eca-109 cells in G0/G1 was increased and that of cells among S phase was reduced with lower doses of Art. However, most of cells were arrested in G2/M phase at the concentration of 100 μmol/L. Art could dramatically inhibit the expression of CDC25A in both mRNA and protein levels. The expression of TGF-β was increased at the same concentration of art. Conclusion Art inhibites the proliferation of tumor cells and modulates cell cycle by down regulation of CDC25A and increasing TGF-β.
出处
《基础医学与临床》
CSCD
北大核心
2007年第2期161-165,共5页
Basic and Clinical Medicine
基金
河北省普通高等学校强势特色学科肿瘤学建设课题
