藏药材纤毛婆婆纳乙酸乙酯提取物的抗癌活性及其机制研究

Study on anti cancer activity and mechanism of ethyl acetate extract from Veronica ciliata Fisch.

摘要:

采用MTT法检测纤毛婆婆纳（Veronica ciliata Fisch.）乙酸乙酯提取物（ethyl acetate extract, VEAE）对五种不同癌细胞（A549, Hela, U20S, MCF-7和SMMC-7721）的增殖抑制作用，筛选出对VEAE最敏感的细胞，并进一步运用DAPI染色、Annexin V/PI 双染及荧光定量PCR（qRT-PCR）来探究其作用机制.MTT结果显示，VEAE对五种癌细胞的增殖均有抑制作用，并呈现时间和剂量的依赖效应， 其中对MCF7细胞的抑制作用最为显著，处理48 h后其IC50达到最低，为63.42±0.19 μg/mL； DAPI染色和Annexin V/PI 双染结果发现，VEAE处理MCF7细胞后，细胞皱缩变圆，凋亡细胞的数量增加，从对照组的5.00%上升到47.45%；qRT-PCR结果表明;VEAE处理MCF7细胞后，凋亡基因 Caspase-3 , Caspase-9 和 Bax 的表达上调，抗凋亡基因〖WTBX〗Bcl-2〖WTBZ〗 的表达下调.由此表明VEAE能抑制癌细胞的增殖，诱导细胞发生凋亡，其机制可能是通过调控与细胞凋亡相关基因的表达来实现.

Abstract:

This study utilized the MTT assay to detect the effect of Veronica ciliata Fisch. ethyl acetate extract (VEAE) on the proliferation of different cancer cells (A549, Hela, U20S, MCF-7, SMMC-7721) and select the most sensitive cells. Furthermore, the mechanism of VEAE inhibiting cancer cells proliferation was measured by DAPI staining, Annexin-V/PI staining and quantitative real-time PCR. MTT assay showed that the proliferation of five cancer cells were gradually suppressed with the increasing of concentration and treatment time of VEAE. And VEAE exhibited the strongest inhibitory effect on MCF-7 cells, and IC50 of VEAE reached to minimum as 63.42±0.19 μg/mL. Further studies by DAPI staining and Annexin-V/PI staining showed that after treatment of MCF-7 cells with VEAE, cell shrinkage and turn round, the ratios of apoptotic cells increased from 5.00% (control group) to 47.45% with the increasing of concentration of VEAE. Quantitative real-time PCR showed that VEAE could promote the expression of Caspase-3, Caspase-9 , Bax genes and inhibit the expression of Bcl-2 gene. It indicated that VEAE could inhibit cells proliferation and induce cells apoptosis. The mechanism may be associated with the expression of apoptosis related genes.