白屈菜碱通过促进p53表达抑制胶质母细胞瘤细胞增殖

Chelidonine inhibits glioblastoma cell proliferation by promoting p53 expression

目的探讨白屈菜碱对胶质瘤母细胞细胞增殖调控的影响及其机制。方法分别以白屈菜碱和二甲基亚砜(DMSO)处理胶质母细胞瘤细胞株U87作为实验组和对照组,细胞计数试剂盒(CCK-8)实验检测U87细胞增殖能力;通过流式细胞术检测细胞周期;通过蛋白质印迹法(Western blot)检测p53和p21蛋白表达;以小RNA干扰技术抑制p53表达;应用GraphPad Prism 6统计分析,组间比较采用Student’s t检验分析。结果白屈菜碱作用于胶质母细胞瘤细胞株后,与对照组(1.343±0.022)比较,实验组细胞活性[吸光度(A)450值(0.783±0.030)]被抑制,差异有统计学意义(t=14.850,P<0.05);细胞周期分析,对照组G1期细胞比例[(54.67±1.76)%],实验组G1期细胞数比例[(65.93±1.61)%]增多,差异有统计学意义(t=4.734,P<0.05),周期阻滞于G1期;Western blot结果显示,与对照组(1.06±0.03、1.07±0.04)比较,实验组p53、p21表达升高(2.78±0.12、4.08±0.13),差异均有统计学意义(t=13.620、22.170,P<0.05)。小RNA干扰技术抑制p53表达后,与对照组[(36.13±0.71)%]比较,实验组细胞G1期细胞[(35.43±0.64)%]比例改变差异无统计学意义(t=0.734,P>0.05),Western blot结果显示,实验组p53、p21表达(1.11±0.01、1.16±0.05)与对照组(1.04±0.03、1.04±0.03)差异均无统计学意义(t=2.209、2.171,P>0.05)。结论白屈菜碱通过促进p53表达抑制胶质母细胞瘤细胞增殖。

Objective To investigate the effect and mechanism of chelidonine on the proliferation of glioblastoma cells.Methods Glioblastoma cell line U87 was treated with chelidonine as the experimental group and dimethylsulfoxide(DMSO)as the control group.Flow cytometry assay was used to detecte cell cycle.Western blotting was used to detecte the protein expressions of p53 and p21.The expression of p53 was inhibited by small RNA interference.GraphPad Prism 6 was used for statistical analysis.The measurement data were expressed as Mean±SD,and the comparison between groups was analyzed by Student’s t test.Results Treated by chelidonine,the proliferation ability(450 A value)(0.783±0.030)of the U87 cells was significantly inhibited compared with the control group(1.343±0.022,t=14.850,P<0.05).Cell cycle analysis showed that the proportion of G1 phase cells in the experimental group(65.93±1.61)%was more than that in the control group[(54.67±1.76)%,t=4.734,P<0.05].Western blotting results showed that compared with the control group(1.06±0.03,1.07±0.04),the expressions of p53 and p21 in the experimental group were significantly increased(2.78±0.12,4.08±0.13,t=13.620,22.170,P<0.05).When the expression of p53 was inhibited,compared with the control group[(36.13±0.71)%],there was no statistically significant difference of the proportion of G1 phase cells in the experimental group[(35.43±0.64)%,t=0.734,P>0.05].Western blotting results showed that compared with the control group(1.04±0.03,1.04±0.03),the expression of p53 and p21 in the experimental group(1.11±0.01,1.16±0.05)showed no significant difference(t=2.209,2.171,P>0.05).Conclusion Chelidonine inhibits glioblastoma cells proliferating by promoting the expression of p53.