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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(SiO_2)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 SiO_2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SiO_2、鬼臼毒素、4′-去甲基表鬼臼毒素和联合用药(12.500、1.250、0.125μg/m L纳米SiO_2分别搭载6.25μg/m L 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 SiO_2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0.125μg/m L纳米SiO_2搭载6.25μg/m L 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 SiO_2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 of podophyllotoxin, 4’-demethylepipodophyllotoxin and its combination with different proportions of nano-silica (SiO_2) on the proliferation of human cervical cancer HeLa cells in vitro and its mechanism. Methods A 25 nm SiO 2 sample was prepared by hydrolysis of tetraethyl orthosilicate. After surface modification, the sample was loaded with 4’-demethyl epipodophyllotoxin. The contents of SiO 2, podophyllotoxin, 4’-demethyl group Podophyllotoxin and combination therapy (12.500,1.250,0.125μg / m L nano-SiO 2 were loaded with 6.25μg / m L 4’-demethylation of epipodophyllotoxin) on HeLa cell proliferation in vitro inhibition; Hoechst 33342 staining assay 25 nm SiO 2, podophyllotoxin and 4’-demethylepipodophyllotoxin on cell apoptosis. The effect of combination therapy on cell morphology was observed by inverted microscope. The effect of 4’-demethyl Effects of epipodophyllotoxin and combination therapy on the expression of apoptosis - related proteins in HeLa cells. Results The inhibitory effect of 4’-demethylepipodophyllotoxin on HeLa cells was better than that of podophyllotoxin. The inhibitory effect of 4’-demethylated epipodophyllotoxin on HeLa cells was better than that of single 4’-demethylated epipodophyllotoxin and 0.125μg / m L nano-SiO 2 carrying 6.25 μg / m L 4’-demethylepipodophyllotoxin, the most obvious inhibition; podophyllotoxin, 4’-demethylepipodophyllotoxin and combination therapy can induce apoptosis ; 4’-demethylepodophyllotoxin and combination therapy can up-regulate the Bax / Bcl-2 ratio and the expression of Caspase-3, p53 and p38. Conclusion The combination of SiO 2 and HeLa cells is more effective than single 4’-demethylepipodophyllotoxin in inhibiting the proliferation of HeLa cells. The mechanism may be through the influence of Bcl-2, Bax, Caspase-3, p53, Protein expression induces apoptosis.