[关键词]
[摘要]
目的 探讨紫草素对人结肠癌SW480细胞凋亡和自噬的影响及其机制。方法 取对数生长期人结肠癌SW480细胞,设对照组(DMSO)、紫草素(0.3、0.5、0.7 μg/mL)和LY294002(PI3K特异性抑制剂,5 μg/mL)组。药物干预48 h后,四甲基偶氮唑盐(MTT)法检测SW480细胞增殖抑制率,Annexin V-FITC流式细胞术分析细胞凋亡状况,蛋白免疫印迹法(Western blotting)检测p-PI3K、p-Akt、p-mTOR、Caspase-3、cleaved Caspase-3、Bcl-2、Bax、LC3蛋白表达并计算Bax/Bcl-2和LC3-II/LC3-I值。结果 与对照组比较,经紫草素0.3、0.5、0.7 μg/mL或LY294002 5 μg/mL干预能够显著提高人结肠癌SW480细胞增殖抑制率和凋亡率(P<0.01);经紫草素0.5、0.7 μg/mL或LY294002 5 μg/mL干预能够显著下调p-PI3K、p-Akt、p-mTOR、Bcl-2蛋白表达,并上调Caspase-3、cleaved Caspase-3、Bax蛋白表达(P<0.05、0.01),提高Bax/Bcl-2和LC3-II/LC3-I值(P<0.01)。与LY294002组比较,经紫草素0.7 μg/mL干预能够显著提高SW480细胞增殖抑制率和凋亡率(P<0.05、0.01),下调p-PI3K、p-Akt、p-mTOR蛋白表达并上调Caspase-3、cleaved Caspase-3、Bax蛋白表达(P<0.05、0.01),提高Bax/Bcl-2和LC3-II/LC3-I值(P<0.01)。结论 紫草素能够促进人结肠癌SW480细胞凋亡和自噬,作用机制可能与抑制PI3K/Akt/mTOR信号通路活化有关。
[Key word]
[Abstract]
Objective To investigate the effect and mechanism of shikonin on apoptosis and autophagy of human colon cancer SW480 cells. Methods The human colon cancer SW480 in logarithmic growth phase was treated with DMSO (control group), shikonin (0.3, 0.5, 0.7 μg/mL) and LY294002 (PI3K-specific inhibitor, 5 μg/mL). Totally 48 h after the drugs were given, the proliferation inhibition rate of the SW480 cells was detected by MTT method, the analysis of apoptotic status and calculation of apoptotic rate were detected by Annexin V-FITC staining flow cytometry, the expression of p-PI3K, p-Akt, p-mTOR, Caspase-3, cleaved Caspase-3, Bax, Bcl-2, LC3 proteins were detected by Western blotting, the ratio of Bax/Bcl-2 and LC3-II/LC3-I were calculated. Results Compared with control group, intervented by shikonin 0.3, 0.5, 0.7 μg/mL or LY294002 5 μg/mL could increase SW480 cells proliferation inhibition rate and apoptosis rate (P<0.01); intervented by shikonin 0.5, 0.7 μg/mL or LY294002 5 μg/mL could down-regulate the expression of p-PI3K, p-Akt, p-mTOR and up-regulate the expression of Caspase-3, cleaved Caspase-3, Bax (P<0.05 or 0.01), increase the ratio of Bax/Bcl-2 and LC3-II/LC3-I (P<0.01). Compared with LY294002 5 μg/mL group, intervented by shikonin 0.7 μg/mL could increase SW480 cells proliferation inhibition rate and apoptosis rate (P<0.05 or 0.01); down-regulate the expression of p-PI3K, p-Akt, p-mTOR and up-regulate the expression of Caspase-3, cleaved Caspase-3, Bax (P<0.05 or 0.01); increase the ratio of Bax/Bcl-2 and LC3-II/LC3-I (P<0.01). Conclusions Shikonin can promote apoptosis and autophagy of human colon cancer SW480 cells; which mechanism may be related to inhibiting the activation of PI3K/Akt/mTOR signaling pathway.
[中图分类号]
R285.5
[基金项目]
河北省医学科学研究课题计划(20191801)