银杏内酯B诱导人结直肠癌HCT116细胞凋亡的研究

Ginkgolide B Induces the Apoptosis of Human Colorectal Cancer HCT116 Cells

目的探讨银杏内酯B(ginkgolide B,GB)对人结直肠癌细胞增殖和凋亡的影响及其可能机制。方法通过制备HCT116裸鼠瘤模型,观察GB对肿瘤生长的影响,并通过体外培养人结直肠癌HCT116细胞,分别加入不同质量浓度GB,与其共孵育72h,噻唑蓝(MTT)法检测细胞增殖抑制率,Giemsa染色观察细胞形态,Annexin V/PI双染法检测细胞凋亡,Western blot法检测Bcl-2蛋白(B-cell leukemia/lymphoma 2,Bcl-2)、Bax蛋白(Bcl-2-associated protein x,Bax)和线粒体融合蛋白1(mitofusin 1,Mfn1)的表达。结果GB抑制裸鼠瘤的生长(P<0.05),并显著抑制HCT116细胞的增殖(P<0.01),诱导HCT116细胞发生凋亡。同时,GB能够显著下调HCT116细胞Mfn1蛋白和Bcl-2蛋白的表达水平(P<0.01),并上调Bax蛋白的表达(P<0.05)。结论 GB可能通过抑制Mfn1的表达,调节促凋亡蛋白Bax活性,降低抗凋亡蛋白Bcl-2的表达,促进结直肠癌细胞的凋亡。

Objective To investigate the effect of ginkgolide B（GB）on proliferation and apoptosis of HCT116 cells and its possible mechanism.Methods The effect of GB on tumor growth was observed in HCT116 nude mice.Furthermore,the HCT116 cells were cultured in vitro and incubated with different concentrations of ginkgolide B for72 h.MTT was used to check the inhibition ratio,cellular morphology was assessed by Giemsa staining,cell apoptosis was evaluated by Annexin V/PI,and the expression of Bcl-2,Bax and Mfn1 was detected by Western blot.Results GB could significantly inhibit the tumor growth of nude mice and the HCT116cells′proliferation（P〈0.05）,and induce HCT116 cell apoptosis in a concentration-dependent manner.The expression of Mfn1 and Bcl-2was remarkably decreased by GB in HCT116cells（P〈0.01）.Meanwhile,the expression of Bax was prominently promoted by GB（P 〈0.05）.Conclusion GB might promote the apoptosis of HCT116 cells to inhibit the proliferation through regulating the activity of Bcl-2,Bax and Mfn1 to suppress the growth of tumor.