目的 研究鬼臼毒素、4'-去甲基表鬼臼毒素及其與不同比例納米二氧化硅（SiO₂）的聯(lián)合用藥對人宮頸癌HeLa細(xì)胞的體外增殖抑制作用，并探討其機(jī)制。方法 采用正硅酸乙酯水解法制備25 nm SiO₂樣品，將其進(jìn)行表面改性后搭載4'-去甲基表鬼臼毒素；MTT法檢測SiO₂、鬼臼毒素、4'-去甲基表鬼臼毒素和聯(lián)合用藥（12.500、1.250、0.125 μg/mL納米SiO₂分別搭載6.25 μg/mL 4'-去甲基表鬼臼毒素）對HeLa細(xì)胞體外增殖的抑制作用；Hoechst 33342染色法檢測25 nm SiO₂的細(xì)胞相容性、鬼臼毒素和4'-去甲基表鬼臼毒素對細(xì)胞凋亡影響；倒置顯微鏡觀察聯(lián)合用藥對細(xì)胞形態(tài)的影響；Western blotting技術(shù)檢測4'-去甲基表鬼臼毒素和聯(lián)合用藥對HeLa細(xì)胞凋亡相關(guān)蛋白表達(dá)的影響。結(jié)果 4'-去甲基表鬼臼毒素對HeLa細(xì)胞的增殖抑制作用優(yōu)于鬼臼毒素，聯(lián)合用藥對HeLa細(xì)胞的抑制作用優(yōu)于單一4'-去甲基表鬼臼毒素，且0.125 μg/mL納米SiO₂搭載6.25 μg/mL 4'-去甲基表鬼臼毒素時(shí)，抑制作用最明顯；鬼臼毒素、4'-去甲基表鬼臼毒素和聯(lián)合用藥均可誘導(dǎo)細(xì)胞凋亡；4'-去甲基表鬼臼毒素和聯(lián)合用藥可以上調(diào)Bax/Bcl-2比值及Caspase-3、p53、p38的表達(dá)水平。結(jié)論 SiO₂聯(lián)合用藥對HeLa細(xì)胞的增殖抑制作用優(yōu)于單一4'-去甲基表鬼臼毒素，其作用機(jī)制可能為，通過影響B(tài)cl-2、Bax、Caspase-3、p53、p38等凋亡相關(guān)蛋白表達(dá)誘導(dǎo)細(xì)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.

甘肅省高等學(xué)?？蒲许?xiàng)目（2013B-059）