小白菊内酯对EC9706细胞增殖及VEGF、NF-κB P65蛋白表达的影响

Effects of parthenolide on expressions of VEGF,NF-κB P65 protein and proliferation of EC9706 cells

目的:探讨小白菊内酯(PTL)对食管癌EC9706细胞的增殖、血管内皮生长因子(VEGF)及核转录因子-κBP65(NF-κBP65)蛋白表达的影响。方法:倒置显微镜下观察不同浓度(20、40和80μmol/L)的PTL作用48h后EC9706细胞的形态学变化;MTT法检测20、40和80μmol/LPTL及20μmol/LPTL与25mg/L5-氟尿嘧啶(5-FU)联用对EC9706细胞增殖的影响;免疫细胞化学法检测20和40μmol/L的PTL作用72h后EC9706细胞VEGF和NF-κBP65蛋白表达的改变。结果:①PTL作用EC9706细胞48h后,细胞数量减少,细胞固缩变圆,核浓缩、核质分布不清,细胞溶解碎裂。②PTL及联合组作用于EC9706细胞24、48和72h后,相同时间段各组细胞增殖抑制率差异均有统计学意义(χ224h=31.420,P<0.001;χ428h=34.590,P<0.001;χ272h=38.190,P<0.001)。PTL对EC9706细胞的增殖有抑制作用;细胞培养72h后PTL与5-FU联合组对EC9706细胞的抑制作用与50mg/L5-FU组相当。③PTL作用EC9706细胞后,与对照组比较VEGF和NF-κBP65蛋白表达均降低(χ2NF-κBP65=25.120,P<0.001;χV2EGF=22.190,P<0.001)。结论:PTL能抑制EC9706细胞的增殖,增加EC9706对5-FU的敏感性;PTL的抗肿瘤机制可能与抑制NF-κB传导通路有关。

Aim：To investigate the effects of parthenolide（PTL） on expressions of vascular endothelium growth factor（VEGF）,nuclear transcription factor-kappaB P65 （NF-κB P65） proteins and proliferation of EC9706 cells in vitro.Methods：Morphologic changes of EC9706 cells were observed by inverted microscope after being treated with 20,40,and 80μmol/L PTL for 48 h.The effects of different concentrations of PTL（20,40 and 80 μmol/L） and 20 μmol/L PTL combined with 25 mg/L 5-fluorouracil （5-FU） on EC9706 cells proliferation were examined by MTT assay.The expressions of VEGF and NF-κB P65 protein of EC9706 cells treated by 20 and 40 μmol/L PTL for 72 h were evaluated by immunocytochemistry method.Results：①After PTL treatment for 48 h,the numbers of EC9706 cells reduced,cells became round and showed cell pyknosis,nucleus concentration,fragment,karyoplasm unclear.②There was significant difference in cell proliferation inhibitory rate among different concentration groups（χ224 h=31.420,P=0.001;χ248 h=34.590,P0.001;χ272 h=38.190,P0.001）.The proliferative activities of EC9706 cells treated with PTL were suppressed.After 72 h,the inhibitory effects in 20 μmol/L PTL combined with 5-FU 25 mg/L group were equivalent with those in 50 mg/L 5-FU group.③The expressions of VEGF and NF-κB P65 protein in EC9706 cells decreased significantly after treatment with PTL（χ2NF-κB P65=25.120,P0.001;χ2VEGF=22.190,P0.001）.Conclusion：PTL could repress EC9706 cellular proliferation activity in vitro,and enhance the sensitivity of EC9706 cells to 5-FU.The anti-tumor mechanisms of PTL could be related to inhibition of NF-κB transduction pathway.