和厚朴酚促进组织因子途径抑制物-2表达并诱导神经胶质瘤细胞凋亡

Promotion of tissue factor pathway inhibitor-2 expression and induction of glioma cell apoptosis by honokiol

目的探究和厚朴酚对神经胶质瘤细胞凋亡的作用及组织因子途径抑制剂因子TFPI-2在其中的分子机制。方法在体外培养神经胶质瘤细胞系U87b,使用不同浓度(0、40、80μM)的和厚朴酚处理神经胶质瘤细胞,利用流式细胞术检测细胞凋亡情况,利用western blot检测TFPI-2、BAX、BCL-2、Cleaved caspase3的蛋白表达情况。利用siRNA沉默U87细胞中TFPI-2的表达,进一步确定TFPI-2的作用。结果 40μM和厚朴酚处理诱导的U87细胞凋亡率(26. 3±0. 55)%明显高于对照组(4. 8±0. 22)%,P <0. 001);相比于对照组(4. 8±0. 22)%,80μM和厚朴酚[(79. 1±0. 31)%,P <0. 001]同样也显著增加细胞凋亡;和厚朴酚成浓度依赖性增加U87细胞内凋亡相关基因BAX/BCL-2比值、Cleaved caspase3的表达以及TFPI-2的蛋白表达;和厚朴酚+si TFP1-2组的细胞凋亡率(12. 7±0. 29)%明显低于和厚朴酚+NC组细胞凋亡率[(57. 2±0. 55)%,P <0. 001];TFPI-2沉默能明显抑制促凋亡蛋白BAX、Cleaved caspase3的表达水平,并促进抑凋亡蛋白BCL-2的表达。结论和厚朴酚可以通过诱导U87神经胶质瘤细胞中TFPI-2高表达,进而促进细胞凋亡的发生。

Objective To explore the effect of honokiol on glioma cell and its cellular mechanism with tissue factor pathway inhibitor-2(TFPI-2)involved.Methods The glioma cell line U87 b was cultured in vitro,with honokiol-treated glioma cells,at different concentrations(0,40,80μM),using flow cytometry,the cell apoptosis was detected.Using western blotting test,the protein expressions of TFPI-2,BAX,BCL02 and cleaved caspase-3 were determined.Using siRNA,the expression of TFPI-2 in U87 cells was silenced.Results Induced by honokion,at the concentration of 40μM,U87 apoptosis(26.3±0.55)%was higher obviously than the control group(4.8±0.22)%,P<0.001.Compared with the control group(4.8±0.22)%,the apoptosis with the induction of 80μM honokiol(79.1±0.31)was increased significantly(P<0.001).BAX/BCL-2 ratio,cleaved caspase-3 expression and TFPI-2 expression were increased in dependence with the concentrations of honokiol.The apoptosis in the group of honokiol+si TFPI-2(12.7±0.29)%was lower obviously than the group of honokiol and NC(57.2±0.55)%,P<0.001.TFPI-2 silencing obviously inhibited the expressions of proapoptotic protein BAX and cleaved caspase-3,promoted the expression of the inhibitory protein BCL-2.Conclusion Honokiol promotes cell apoptosis through inducing TFPI-2 expression in U87 glioma cells.