人参皂苷IH901对ECV304细胞增殖和迁移的影响及其分子机制

Inhibitory effect of ginseng saponin IH901 on proliferation and metastasis of ECV304 cell line and its molecular mechanism

本文探讨人参皂苷20-O-(β-D-吡喃葡萄糖)-20(S)-原人参二醇皂苷(IH901)对ECV304细胞的增殖和迁移的抑制作用。以不同浓度的IH901体外作用ECV304细胞,采用MTT法检测IH901对ECV304细胞生长的抑制效果和对基质黏附能力的影响,显微镜观察细胞形态学变化,流式细胞术检测细胞周期变化和细胞凋亡率,划痕法检测细胞的迁移能力,ELISA检测试剂盒检测VEGF和bFGF在ECV304细胞中的表达,Western blotting检测被激活的cleaved caspase-9和cleaved caspase-3两种蛋白的表达量。结果表明,人参皂苷IH901可能通过抑制ECV304细胞分泌VEGF和bFGF,并上调促凋亡蛋白cleaved caspase-9和cleaved caspase-3的表达,从而使得细胞被阻滞在G0/G1期,并且在形态学上观察到典型的细胞凋亡变化,如细胞皱缩,体积变小,伴随核固缩,最终导致人参皂苷IH901对ECV304细胞与细胞外基质黏附能力和细胞迁移能力有明显的抑制作用,且呈时间和浓度依赖关系。

This study aims to investigate the inhibitory effect on proliferation and metastasis of 20-O-（β-D-glucopyranosyl）-20（S）-protopanaxadiol （IH901） on ECV304 cell line. MTT assay was used to examine the effect of cell proliferation inhibition and the adhesive ability of ECV304 cells to artificial basement membrane. Morphology of cell apoptosis was observed with phase contrast microscope. Apoptosis rate and cell cycle were detected by flow cytometry （FCM）. Cell migration was measured by wound healing assay. ELISA kit was used to detect VEGF and bFGF. Caspases were detected by Western blotting. Results indicated that ginseng saponin IH901 can downregulate the expression of growth promoting protein VEGF and bFGF, and upregulate proapoptosis protein cleaved caspase-9 and cleaved caspase-3. The increase in the apoptotic sub-G1 fraction is in a dose-dependent manner, and cell cycle arrests in the G0/G1 phase was detected by FCM. Morphological examination of IH901-treated samples showed cells with chromatin condensation, cell shrinkage, and all typical characteristics of apoptotic cells. Therefore, IH901 dramatically suppresses cell proliferation and adhesion and migration of ECV304 cell line.