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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 inhibitory effect of artesunate (Art) on human esophageal cancer Eca-109 cell line, and further explore the relationship between Art induced tumor cell cycle arrest and the expression of CDC25A and TGF-β. Methods Human esophageal cancer Eca-109 cell line and normal human peripheral blood mononuclear cells (hPBMC) were cultured in vitro. Cell proliferation was measured by MTT assay. Cell cycle was determined by flow cytometry (FCM); CDC25A mRNA expression was detected by RT-PCR. The protein expression was detected by Western blot. Results Art could significantly inhibit the proliferation of Eca-109 cells with IC50 of (68.80±0.76)μmol/L, but had no obvious inhibitory effect on the proliferation of hPBMC. The low concentration of Art can arrest cells in G0/G1 phase, and the number of cells in S phase decreases significantly. When the concentration reaches 100 μmol/L, cells arrest in G2/M phase. Art can significantly inhibit the expression of CDC25A mRNA and protein in Eca-109 cells and significantly up-regulate the expression of TGF-β protein. Conclusion Art can inhibit the growth of tumor cells, up-regulate the expression of TGF-β and inhibit the expression of CDC25A.