石上柏乙酸乙酯萃取物对喉癌Hep-2细胞增殖与迁移作用的研究

摘要

本文利用三种不同极性溶剂从石上柏中分离出活性成分,观察其对喉癌Hep-2细胞的增殖与迁移作用的影响.干燥的石上柏全草粉碎后经乙醇回流提取浓缩得浸膏,依次采用石油醚、二氯甲烷、乙酸乙酯三种溶剂进行萃取,以获得各极性溶剂萃取部位;MTT法检测石上柏提取物对Hep-2细胞增殖的影响;流式细胞仪检测石上柏提取物对Hep-2细胞周期的影响;细胞划痕实验检测石上柏提取物对Hep-2细胞迁移能力的影响;采用p53报告基因探讨石上柏提取物对喉癌Hep-2细胞的作用机制.结果显示31.2～500.0 μg/mL的乙酸乙酯萃取部位能明显抑制Hep-2细胞的生长及划痕区域的迁移;15.0 ～ 60.0 μg/mL的乙酸乙酯萃取物部位能使喉癌Hep-2细胞阻滞于G1期,并且能激活p53的表达.结果表明石上柏乙酸乙酯萃取部位可明显抑制喉癌Hep-2细胞的生长及迁移,其作用与激活p53通路有关.%In this study,Selaginella doederleinii was extracted by three reagents with different hydrophobicity.The effects of these extracts on proliferation,cell cycle and migration of Hep-2 cells in vitro were assessed.The petroleum ether,dichloromethane and ethyl acetate fractions were prepared from ethanol extract of S.doederleinii.The effect of these fractions on proliferation of Hep-2 cells was determined by MTT assay;cell cycle was measured by flow cytometry (FCM);the anti-migration ability was observed through cell scratch method;and the molecular mechanism was determined by p53 reporter gene.The results showed that the ethyl acetate extract (31.2-500.0 μg/mL) showed the best anti-tumor activity in vitro and it can induce the inhibition of cell migration.Ethyl acetate extract (15.0-60.0 μg/mL) also can induce cell cycle arrest and activate the expression of p53.It indicated that the ethyl acetate extract of S.doederleinii can significantly inhibit the growth and migration of Hep-2 cells through a p53 pathway.