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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 on proliferation and apoptosis-inducing effect of human osteosarcoma cell line MG-63. Methods The changes of cell cycle were detected by flow cytometry. The expressions of CyclinD1, P21 and Survivin, Bcl-2 and Bax were detected by Western blotting. The changes of telomerase activity in MG-63 cells were detected by TRAP-PCR-ELISA Z-IETD-fmk blocking test was used to detect the changes of apoptosis of MG-63 cells treated with oridonin before and after the blockade of Caspase-8. Results Oridonin increased the percentage of MG-63 cells in G0 / G1 phase and decreased the percentage of MG-63 cells in S phase. The concentration of Oridonin could inhibit the expression of CyclinD1, Survivin and Bcl-2 in MG-63 cells and upregulate the expression of P21 and Bax , Respectively. Z-IETD-fmk could partially reverse and attenuate the apoptosis-inducing effect of oridonin on MG-63 cells after blocking Caspase-8 activity in a concentration-dependent manner. Conclusion Oridonin can inhibit telomerase activity and activate Caspase-8 by blocking the expression of cell cycle regulatory proteins, blocking the G1 / S phase of the cell cycle, inhibiting Survivin and Bcl-2, up-regulating Bax, Inhibit the proliferation of MG-63 cells and induce the apoptosis of MG-63 cells.