黄连素对体外培养siha细胞增殖的影响

Effect of berberine on proliferation of Siha cells in vitro

目的:观察黄连素(berberine,ber)对人子宫颈癌siha细胞生长的抑制作用及其对凋亡基因caspase-9、caspase-3的影响,以及caspase-3的活化形式cleaved caspase-3含量的变化。初步探讨ber抑制siha细胞增殖的机制。方法:体外培养人宫颈癌siha细胞,将其分为对照组(二甲亚砜)及ber 20、40和80μmol.L-1,实验组,分别用二甲亚砜、ber处理细胞24、48、72和96 h后,经MTT法检测ber对siha细胞增殖的影响;RT-PCR、Westem blotting、免疫细胞化学染色法观察凋亡相关基因caspase-9、caspase-3的转录、蛋白表达情况。结果:实验组MTT值高于对照组,即实验组细胞增殖受到明显抑制;与对照组相比,实验组凋亡相关基因caspase-9、caspase-3的转录水平均上调(P<0.01)、两基因蛋白表达水平明显上调(P<0.01),活化形caspase-3含量增加。结论:ber可抑制体外培养siha细胞增殖,诱导siha细胞发生凋亡。其机制可能与激活caspase家族级联反应有关。

Objective：To observe the inhibiting effect of berberine on proliferation of human cervical cancer Siha cells and the effect on apoptosis related genes（caspase-9 and caspase-3）,and the change of cleaved caspase-3 content,explore the mechanism of inhibiting effect of berberine on proliferation of Siha cells.Methods：Human cervical cancer Siha cells were cultured in vitro,then they were divided into control group（dimethylsulfoxide） and experimental group（20 μmol·L-1 berberine group,40 μmol·L-1 berberine group and 80 μmol· L-1 berberine group）,Siha cells in the two groups were treated with dimethylsulfoxide and berberine for 24,48,72 and 96 hours,respectively,then MTT method was used to detect the effect of berberine on proliferation of Siha cells;RT-PCR,Westem blotting and immunocytochemical staining were used to observe the transcriptions and expressions of caspase-9 protein and caspase-3 protein.Results：MTT value in experimental group was higher than that in control group,the proliferation of Siha cells in experimental group was inhibited significantly;compared with control group,the transcription levels of caspase-9 and caspase-3 were up-regulated（P0.01）,the expression levels of caspase-9 protein and caspase-3 protein were up-regulated（P0.01）,cleaved caspase-3 content increased.Conclusion：Berberine can inhibit the proliferation of Siha cells in vitro and induce apoptosis of Siha cells,the mechanism may be related to activation of caspase family.