臭牡丹总黄酮对A549细胞上皮间质转化相关蛋白的影响

余　娜，马思静，朱克俭*

（1.湖南中医药大学，湖南长沙410208；2.湖南省中医药研究院，湖南长沙410006）

臭牡丹总黄酮对A549细胞上皮间质转化相关蛋白的影响

余娜1，马思静2，朱克俭2*

（1.湖南中医药大学，湖南长沙410208；2.湖南省中医药研究院，湖南长沙410006）

目的观察臭牡丹总黄酮对A549肺腺癌细胞上皮间质转化（epithelial-mesenchymal transition，EMT）相关蛋白的影响。方法构建β-catenin真核过表达载体并转染A549细胞，将细胞分为空载组、β-catenin过表达组、空载组+臭牡丹总黄酮组、β-catenin过表达+臭牡丹总黄酮组。MTT法检测不同浓度的臭牡丹总黄酮对A549细胞体外增殖的抑制作用。实时定量PCR检测各组β-catenin mRNA的表达，Western Blot检测各组β-catenin、E-Cardherin、Vimentin的蛋白水平。结果MTT显示A549细胞活力随臭牡丹总黄酮浓度的增加受到明显抑制，IC50为2.1 mg/mL。β-catenin过表达组较空载组的β-catenin、Vimentin表达均明显上调，E-cardherin表达下调。臭牡丹总黄酮在空载组与β-catenin过表达组均能明显降低βcatenin mRNA与蛋白水平，增加E-cardherin蛋白表达，略微降低Vimentin蛋白表达。结论臭牡丹总黄酮对A549肺癌细胞具有一定的体外抑制增殖作用，其抗肿瘤作用的可能机制是通过调控EMT的相关蛋白从而逆转β-catenin诱导的EMT现象而起作用。

臭牡丹总黄酮；上皮间质转化；β-catenin过表达载体；A549细胞

〔Abstract〕Objective To observe the effect of the total flavonoid of Clerodendron bungei Steud（CMD）on the epithelial mesenchvmal transition related proteins in A549 cells.M ethods Building theβ-catenin overexpression vector and transfected A549 cells.The cells can be divied into empty vector group，β-catenin overexpression group，empty vector group +CMD group，β-catenin overexpression+CMD group.The proliferation inhibition of the total flavonoid of Clerodendron bungei Steud with different concentrations on A549 cells in vitro was detected by MTT method.The expression ofβ-catenin mRNA was determined with real-time PCR，and the protein expression ofβ-catenin，E-Cardherin，Vimentin was detected with Western Blot.Results MTT showed that the A549 cells vitality were significantly inhibited with the increasing concentrations of the CMD，the IC50 was 2.1mg/mL.Compared with the empty vector group，the expression ofβ-catenin and vimentin was significantly higher，the E-cardherin was lower inβ-catenin overexpression group.The total flavonoid of Clerodendron bungei Steud can reduce the expression ofβ-catenin，increase the expression of E-cardherin，and slightly reduce the expression of Vimentin protein in empty vehicle and theβ-catenin overexpression groups.Conclusion the total flavonoid of Clerodendron bungei Steud has a certain proliferation inhibition effect on A549 lung cancer cells in vitro.The possible mechanisms is that，it can regulate some related proteins to reverse the EMT phenomenon throughβ-catenin cell signaling.

〔Keywords〕the total flavonoid of Clerodendron bungei Steud；the epithelial mesenchvmal transition；β-catenin expression vector；A549 cell

原发性支气管肺癌是一种发生于支气管黏膜及腺体的恶性肿瘤，属中医学“肺积”、“肺岩”、“息贲”等范畴，其发病率和死亡率在全球以及我国均居首位［1-2］。其中，肿瘤局部复发和远处转移是肿瘤病程发生发展的主要原因，80%以上的肺癌患者确诊时癌细胞已发生全身侵犯和转移，进入疾病晚期。臭牡丹性平，味辛、苦，有“祛风除湿、解毒散瘀”之功，对多种肿瘤具有临床治疗作用，尤以肺癌为佳。……