美洲商陆抗病毒蛋白对人神经胶质瘤细胞U251细胞增殖和凋亡的影响  

Effects of recombinant pokeweed antiviral proteins from Phytolacca amercana on the proliferation and apoptosis of Human Gliomaous Cells U251

作　　者：向莉[1] 李书剑[1] 张杰文[1] 

机构地区：[1]河南省人民医院神经内科,郑州450003

出　　处：《郑州大学学报（医学版）》2011年第5期742-744,共3页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的:观察美洲商陆抗病毒蛋白(PAP)对人神经胶质瘤细胞U251细胞增殖和凋亡的影响。方法:采用MTT法检测0、20、40、60、80和100mg/LPAP处理48h以及40mg/LPAP处理24、36、48和72h对神经胶质瘤细胞U251细胞生长的影响;40mg/LPAP处理U251细胞36h后,采用荧光染色技术检测细胞凋亡,采用流式细胞仪检测U251细胞周期分布的影响;采用Northernblot和Westernblot检测40mg/LPAP处理24、48、72和96h对U251细胞周期调控蛋白FasL和Fas的影响。结果:0、20、40、60、80和100mg/LPAP处理48h,U251细胞的增殖抑制率间的差异有统计学意义(F=284.560,P=0.007),40mg/LPAP处理24、36、48和72h,U251细胞的增殖抑制率间的差异有统计学意义(F=280.250,P=0.045)。PAP促进U251细胞凋亡(t=106.350,P=0.007),PAP改变细胞周期分布,使G0+G1期细胞比例增高,S期比例降低。PAP还可上调FasL蛋白表达,下调Fas蛋白表达。结论:PAP可改变细胞周期分布,影响细胞周期调控蛋白表达,并可诱导U251细胞凋亡,从而抑制细胞增殖。Aim： To study the effects of pokeweed antiviral protein（PAP） on cell proliferation and apoptosis on U251 cells and to elucidate its molecular mechanism.Methods：The proportion of the periodic tumor cells were altered by 0,20,40,60,80,100 mg/L PAP treated 48 h using MTT and Sub-G1 curves were displayed at 40 mg/L PAP treated 36 h by flow cytometry analysis.With fluorescence staining assay to detect the U251 apoptosis.The mRNAs and protein expression of the FasL and Fas treated at 40 mg/L PAP was examined by Northern blot and Western blot.Results：PAP significantly suppressed U251 cell proliferation by 0,20,40,60,80,100 mg/L PAP treated 48 h（F=284.560,P=0.007） or 40 ng/L PAP treated 24,36,48,72 h （F=28.250,P=0.045）.The proportion of the periodic tumor cells were altered by PAP.PAP decreased the proportion of cells in S phase and increased the proportion of cells in G0/G1 and G2/M phases of the cell cycle.The mRNAs and protein expression of the FasL downregulated gradually as treated time increased and the mRNA and protein expression levels of Fas was upregulated gradually as treated time increased.Conclusion：PAP can induce apoptosis and alert cell cycle arrest of the Human Gliomaous Cells U251 cells.

关 键 词：流式细胞 美洲商陆抗病毒蛋白 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]