臭牡丹总黄酮对A549细胞增殖迁移侵袭力及Wnt通路相关蛋白的影响  

Effect of Total Flavonoids Isolated from Clerodendrum Bungei on Proliferation,Migration,Invasion Ability and Wnt Pathway Associated Protein of A549 Cells

作　　者：余娜[1] 朱克俭[2] 马思静 谭小宁[2] 唐皓[2] Yu Na;Zhu Kejian;Ma Sijing;Tan Xiaoning;Tang Hao(Hunan University of Chinese Medicine, Changsha 410208, China;Hunan Academy of Chinese Medicine, Changsha 410006, China)

机构地区：[1]湖南中医药大学,长沙410208 [2]湖南省中医药研究院,长沙410006

出　　处：《世界中医药》2018年第4期978-983,987,共7页World Chinese Medicine

基　　金：国家自然科学基金项目(81503452)

摘　　要：目的:观察臭牡丹总黄酮对β-catenin过表达A549细胞增殖、迁移、侵袭力以及Wnt通路相关蛋白的影响。方法:选取构建β-catenin真核过表达载体并转染A549细胞,将其分为6组,分别为未转染细胞、未转染细胞+臭牡丹总黄酮、空载组、空载+臭牡丹总黄酮、β-catenin过表达组、β-catenin过表达+臭牡丹总黄酮。MTT法检测各组细胞的相对存活率,划痕实验检测各组细胞迁移能力,Transwell小室侵袭实验检测各组细胞的侵袭力。Western Blot检测各组Wnt通路相关蛋白β-catenin、GSK-3β、P-GSK-3β、c-myc、CyclinD1的表达。结果:转染β-catenin过表达的A549细胞增殖能力、迁移能力、侵袭力均明显增强(P<0.05),并显著上调wnt通路相关因子β-catenin、C-Myc,CyclinD的蛋白表达,下调P-GSK-3β的蛋白表达(P<0.05)。采用臭牡丹总黄酮干预后,能明显降低各组细胞的存活率,降低细胞向划痕区域迁移的能力,减少侵袭小室穿过基膜的细胞数(P<0.05)。并下调β-catenin、C-Myc、CyclinD1表达(P<0.05),上调P-GSK-3β表达(P<0.05)。臭牡丹总黄酮的干预作用对转染β-catenin过表达细胞尤为明显。结论:转染β-catenin过表达的A549细胞其增殖、迁移、侵袭力均明显增强,能激活Wnt/β-catenin通路。臭牡丹总黄酮可抑制A549细胞的增殖、迁移、侵袭力,其机制可能是通过抑制细胞中β-catenin的高表达从而调控下游的一系列因子而起到抑制Wnt/β-catenin通路的作用。Objective： To observe the effect of total flavonoids isolated from clerodendrum bungei on proliferation,migration,invasion of human lung adenocarcinoma A549 cell and its effect on the proteins related to Wnt signal pathway. Methods： β-catenin over-expression plasmid was established and A549 cell was transfected. A549 cells were divided into 6 groups： normal A549 cells,normal A549 cells ＋ total flavonoids isolated from clerodendrum bungei group,empty vector group,empty vector ＋ total flavonoids isolated from clerodendrum bungei group,β-catenin over-expression group,β-catenin ＋ total flavonoids isolated from clerodendrum bungei group. MTT assay detected relative survival rate of the cell,and scratch test observed cell migration. Transwell experiment detected the cell invasion. Western Blot was used to detect proteins of the factors that were related to Wnt pathway： β-catenin,GSK-3β,P-GSK-3β,c-myc,CyclinD1. Results： The proliferation,migration and invasion of A549 cell was enhanced significantly after transfected with β-catenin over-expression plasmid（ P 〈 0. 05 or 0. 01）,along with the growth in number of β-catenin,C-Myc and CyclinD which were related to Wnt signal pathway and the decrease of P-GSK-3β. After interfered with total flavonoids isolated from clerodendrum bungei,survival rate of A549 cell in each group declined significantly and the ability of migration and proliferation was decreased（ P 〈 0. 05）. It down-regulated the expression of β-catenin,C-Myc and CyclinD1（ P 〈 0. 05） and up-regulated the expression of P-GSK-3β（ P 〈 0. 05）. The effect on groups transfected with β-catenin over-expression plasmid were especially obvious. Conclusion： Overexpression of β-catenin could promote the invasion,migration and proliferation in the A549 cell line and activate Wnt/β-catenin pathway,while total flavonoids isolated from clerodendrum bungei could inhibit the invasion,migration and proliferation in the A549 cell line,The pharmacological mechanism is probably in

关 键 词：臭牡丹总黄酮 上皮间质转化 WNT信号通路 β-catenin过表达载体 

分 类 号：R284[医药卫生—中药学]