鸦胆子乙醇提取物对血小板源性生长因子受体介导细胞迁移的抑制作用

Inhibitory effect of ethanol extract from Java Brucea Fruit on cell migration mediated by platelet-derived growth factor receptor β

目的:观察鸦胆子乙醇提取物对血小板源性生长因子受体β(PDGFRβ)介导细胞迁移的影响,阐明其起作用的有效成分。方法:用脂质体介导的细胞转染技术将人PDGFRβ表达载体转染体外培养的不表达PDGFRβ的PAE细胞,G418抗性筛选建立稳定转染的细胞株,采用RT-PCR法检测PDGFRβ的表达;用细胞损伤修复实验观察细胞损伤24 h完全修复所需的血小板源性生长因子BB(PDGFBB)最低浓度;划痕实验观察加入不同浓度鸦胆子乙醇提取物24 h后,计算细胞迁移的抑制率及鸦胆子发挥抑制作用的主要成分溶于乙醇溶液的浓度。结果:RT-PCR检测转染组细胞表达PDGFRβ;细胞损伤修复实验检测细胞损伤24 h完全修复所需的血小板源性生长因子BB(PDGFBB)最低浓度为10μg.L-1;划痕实验检测随着鸦胆子乙醇提取物(70%乙醇溶液)浓度的增加,对细胞迁移的抑制率增加(P<0.05),呈剂量效应关系;50%乙醇提取物和25%乙醇提取物(浓度≤10μg.L-1)对细胞迁移抑制不明显,鸦胆子乙醇提取物浓度>10μg.L-1主要导致细胞死亡。结论:鸦胆子抗肿瘤作用机制可能是通过抑制PDGFRβ介导的细胞迁移来实现的,其发挥作用的主要成分更易溶于70%的乙醇溶液中。

Objective To study the effects of ethanol extract from Java Brucea Fruit on platelet-derived growth factor receptor β（PDGFRβ） mediated cell migration and to obtain the valuable messages on the characteristics of its active ingredient.Methods PAE cells was transfected with the vector expressing human PDGFRβ with Transfectom 2000;After screening by G418 resisitance,RT-PCR was used to monitor the expression of PDGFRβ in the cells;Wound healing of the cells was used to examine the lowest consistency of PDGFBB and inhibitory effect of ethanol extract of Java Brucea Fruit on cell migration after restoring 24 h.Results Human PDGFRβ was stably expressed in PAE cells transfected with the expressing vector.The lowest consistency of exogenous PDGFBB which promoted PDGFRβ mediated cell migration was 10 μg·L^-10% ethanol extract of Java Brucea Fruit which strongly inhibited PDGFRβ mediated cell migration was dose-dependent（P〈0.05）,50% and 25% ethanol extract from Java Brucea Fruit（≤10 μg·L^-1had the weaker inhibitory effect,ethanol extract from Java Brucea Fruit（〉10 μg·L^-1mainly caused the death of the cell.Conclusion Ethanol extract of Java Brucea Fruit has a strongly inhibitory effect on the PDGFRβ mediated cell migration which could play a major role in its effects against metastasis of malignant tumor,the active ingredients of it could be more dissolvable in the 70% ethanol.