摘要
目的研究冬凌草甲素对人骨肉瘤细胞株MG-63细胞增殖抑制和凋亡诱导效应的分子机制。方法以流式细胞仪检测细胞周期变化;Western印迹检测CyclinD1、P21以及凋亡相关蛋白Survivin、Bcl-2和Bax表达的变化;TRAP-PCR-ELISA方法检测MG-63细胞端粒酶活性的变化;Z-IETD-fmk阻断试验检测Caspase-8阻断前后,冬凌草甲素对MG-63细胞凋亡影响的变化。结果冬凌草甲素增加G0/G1期MG-63细胞百分率,降低S期MG-63细胞百分率;浓度依赖性抑制MG-63细胞的CyclinD1、Survivin和Bcl-2蛋白表达,上调P21和Bax蛋白表达;浓度依赖性抑制MG-63细胞端粒酶的活性;Z-IETD-fmk阻断Caspase-8活性后,能部分逆转和减弱冬凌草甲素对MG-63细胞的凋亡诱导作用。结论冬凌草甲素通过影响细胞周期调节蛋白、阻断细胞周期G1/S期"稽查点";抑制Survivin、Bcl-2,上调Bax;抑制端粒酶活性以及活化Caspase-8等途径抑制MG-63细胞增殖及诱导MG-63细胞凋亡。
Objective To study the molecular mechanism of oridonin in the inhibiting the proliferation and inducing apoptosis of osteosarcoma cell MG-63. Methods The change of cell cycle was analyzed by flow cytometry, the pro- tein expression of CyclinD~, P21, and apoptosis-associated protein Survivin, Bcl-2 and Bax in MG-63 ceils were detected by Western blotting method. TRAP-PCR-ELISA was applied for the detection of telomerase activities. After the activation of Caspase-8 was inhibited, the proliferation-inhibiting and apoptosis-inducing effects of Oridonin were detected by Z-IETD-fmk blocking test. Results Oridonin increased the percentage of G0/G1 phase and decreased S phase of MG-63 cells, down-regulated CyclinD1, Survivin and Bcl-2, and up-regulated P21 and Bax protein in a concentration-dependent way in MG-63 cells. Oridonin down-regulated telomerase activities in a concentration-de- pendent way in MG-63 cells. The apoptosis-inducing effects of oridonin were partly suppressed after Z-IETD-fmk blocking the Caspase-8 activation of MG-63 cells. Conclusion Oridonin inhibits proliferation and induces apoptosis of MG-63 cells through regulating cell cycle protein, blocking checkpoint of G~/S phase, down-regulating Survivin and Bcl-2, up-regulating Bax and activating Caspase-8.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2013年第1期43-47,共5页
Traditional Chinese Drug Research and Clinical Pharmacology
