目的 研究鬼臼毒素、4'-去甲基表鬼臼毒素及其与不同比例纳米二氧化硅（SiO₂）的联合用药对人宫颈癌HeLa细胞的体外增殖抑制作用，并探讨其机制。方法 采用正硅酸乙酯水解法制备25 nm SiO₂样品，将其进行表面改性后搭载4'-去甲基表鬼臼毒素；MTT法检测SiO₂、鬼臼毒素、4'-去甲基表鬼臼毒素和联合用药（12.500、1.250、0.125 μg/mL纳米SiO₂分别搭载6.25 μg/mL 4'-去甲基表鬼臼毒素）对HeLa细胞体外增殖的抑制作用；Hoechst 33342染色法检测25 nm SiO₂的细胞相容性、鬼臼毒素和4'-去甲基表鬼臼毒素对细胞凋亡影响；倒置显微镜观察联合用药对细胞形态的影响；Western blotting技术检测4'-去甲基表鬼臼毒素和联合用药对HeLa细胞凋亡相关蛋白表达的影响。结果 4'-去甲基表鬼臼毒素对HeLa细胞的增殖抑制作用优于鬼臼毒素，联合用药对HeLa细胞的抑制作用优于单一4'-去甲基表鬼臼毒素，且0.125 μg/mL纳米SiO₂搭载6.25 μg/mL 4'-去甲基表鬼臼毒素时，抑制作用最明显；鬼臼毒素、4'-去甲基表鬼臼毒素和联合用药均可诱导细胞凋亡；4'-去甲基表鬼臼毒素和联合用药可以上调Bax/Bcl-2比值及Caspase-3、p53、p38的表达水平。结论 SiO₂联合用药对HeLa细胞的增殖抑制作用优于单一4'-去甲基表鬼臼毒素，其作用机制可能为，通过影响Bcl-2、Bax、Caspase-3、p53、p38等凋亡相关蛋白表达诱导细胞凋亡。

Objective To study the inhibitory effect on proliferation of Hela cells of podophyllotoxin, 4'-demethylepi-podophyllotoxin and drug combination of different proportion of nano-sillca (SiO₂) and 4'-demethylepi-podophyllotoxin in vitro, and discuss the mechanism. Methods Used ethyl silicate hydrolysis method to prepare 25 nm SiO₂ sample, next carried 4'-demethylepi-podophyllotoxin after the surface modification, and measure cell campatibility by MTT method and Hoechst 33342. The inhibitory effect of podophyllotoxin, 4'-demethylepi-podophyllotoxin and drug combination on proliferation of Hela cells was measured by MTT assay. Hoechst 33342 staining method was used to detect cell apoptosis. The effect of drug combination treatment on cell morphology was observed by inverted microscope. Western blotting technique was used to detected effect of 4'-demethylepi-podophyllotoxin and drug combination on expression of apoptosis related protein. Results Inhibitory effect onproliferation of Hela cells of 4'-demethylepi-podophyllotoxin is superior to podophyllotoxin, inhibitory effect of drug combination is superior to the single 4'-demethylepi-podophyllotoxin, the inhibition of drug combination with 0.125 μg/mL nano SiO₂ and 6.25 μg/mL 4'-demethylepi-podophyllotoxin is the most obvious. MTT and Hoechst 33342 experimental results showed that the 25 nm SiO₂ have good cell compatibility. Podophyllotoxin, 4'-demethylepi-podophyllotoxin and drug combination can induce apoptosis. Western blotting results showed that 4'-demethylepi-podophyllotoxin and drug combination can up-regulate the ratio of Bax/Bcl-2 and the expression level of Caspase-3、P53 and P38. Conclusion In vitro experimental performance of drug combination is superior to single 4'-demethylepi-podophyllotoxin, it is may by effecting the expression of Bcl-2, Bax, Caspase-3, P53, and P38 and others apoptosis related protein to induce Hela cell apoptosis.

甘肃省高等学校科研项目（2013B-059）