【摘要】 目的:探讨天花粉蛋白(TCS)对食管癌Eca-109细胞增殖的影响及其作用机制。方法:采用MTT法检测TCS对Eca-109细胞增殖的抑制率;电子显微镜和AnnexinV-FITC/PI双染检测细胞凋亡;流式细胞仪分析细胞周期;RT-PCR法检测Eca-109细胞Survivin mRNA表达变化;Western blot检测Survivin、Caspase-3与C-myc蛋白表达变化;TRAP-PCR银染法定性测定细胞的端粒酶活性。结果:TCS对Eca-109细胞增殖有抑制作用且具有剂量-时间效应;TCS明显诱导细胞凋亡,药物组凋亡率高于对照组(P<0.05),并且药物组的S期细胞明显高于对照组(P<0.05);TCS可降低Survivin mRNA和蛋白表达,增加Caspase-3蛋白表达,下调C-myc蛋白表达、使端粒酶活性降低。结论:TCS能够抑制Eca-109细胞增殖。其机制可能为TCS影响了细胞周期和抑制了Survivin表达。更多还原

【Abstract】 Objective:To investigate the effects of trichosanthin(TCS) on the proliferation of human esophageal carcinoma Eca -109 cells and its mechanisms there of .Methods:Cell growth inhibition of TCS on Eca -109 cells was analyzed by MTT assay. Cell apoptosis was detected by electronic microscopy and cytometry by annexinV-FITC/PI double stain. Cell cycle was analyzed by cytometry. The mRNA expression of survivin was studied by RT-PCR. Protein expression of survivin, Caspase-3 and C-myc were studied by Western blot. Telomelse activity was determined by TRAP-PCR kit. Results: TCS could inhibit the proliferation of Eca-109 cells in a time and dose dependant manner. TCS induced cell apoptosis. The cell apoptosis rate in TCS group was significantly higher than that in control group (P < 0.05). The S stage cell percentage in TCS group was signifi- cantly higher than that in control group (P < 0.05). TCS decreased mRNA and protein expressions of survivin, increased protein expressions of caspase-3, down-regulate the expression of C-myc, and decreased the telomelse activity. Conclusion: TCS can inhibit Eca-109 cell proliferation. The possible mechanism is that TCS can influence cell cycle and inhibit the expression of survivin gene.更多还原