摘要
目的观察补中益气汤含药血清对转化生长因子(TGF)-β1诱导前后,A549/DDP细胞Snail、E-钙黏蛋白(cadherin)表达的影响,探讨补中益气汤对A549/DDP细胞上皮-间充质转化(EMT)的干预作用。方法A549/DDP细胞分为:空白组、TGF-β1组(TGF-β15 ng/L,48 h)、中药组(10%补中益气汤含药血清)、干扰组(Snail干扰)。采用siRNA技术,运用免疫细胞化学法、Western印迹法、实时荧光定量PCR法,检测补中益气汤含药血清对A549/DDP细胞Snail、E-cadherin及mRNA水平的影响。结果与空白组比较,TGF-β1组E-cadherin及mRNA表达水平显著下调(P<0.05);与TGF-β1组比较,中药组及干扰组E-cadherin及mRNA表达水平显著上调(P<0.05)。而Snail蛋白及mRNA水平的影响呈现相反的变化趋势。结论Snail作为转录因子,可抑制E-cadherin基因的表达,诱导EMT发生,而补中益气汤含药血清可通过干预Snail蛋白及基因的表达,解除其对E-cadherin的抑制作用,从而逆转A549/DDP细胞EMT。
Objective To observe the expression of Snail and E-cadherin in cell line A549/DDP before and after the serum containing Buzhong Yiqi Decoction induced transforming growth factor(TGF)-β1,and to explore the intervention of Buzhong Yiqi Decoction with epithelial-mesenchymal transition(EMT).Methods A549/DDP cells were divided into blank group,TGF-β1 group(TGF-β15 ng/L,48 h),tradtional Chinese medicine group(10%serum containing Buzhong Yiqi Decoction),interference group(Snail interference).siRNA technology,immunocytochemistry,Western blot and real-time fluorescence quantitative PCR were used to detect serum containing Buzhong Yiqi Decoction on the influence of Snail,E-cadherin and mRNA levels in A549/DDP cell.Results Compared with blank group,the E-cadherin and mRNA expression levels in TGF-β1 group were significantly decreased(P<0.05).Compared with TGF-β1 group,the E-cadherin and mRNA expression levels in tradtional Chinese medicine group and interference group were significantly increased(P<0.05).Snail protein and mRNA levels showed the opposite trend.Conclusions As a transcription factor,Snail could inhibit E-cadherin gene expression and induce the occurrence of EMT.Serum containing Buzhong Yiqi Decoction could relieve the inhibition of E-cadherin by intervening Snail protein and gene expressions,and reverse the A549/DDP cell EMT.
作者
于丹
王莹
高原
刘春英
YU Dan;WANG Ying;GAO Yuan(Collage of Integrative Chinese and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)
出处
《中国老年学杂志》
CAS
北大核心
2023年第7期1636-1639,共4页
Chinese Journal of Gerontology
基金
国家自然科学基金资助项目(No.81973735)
辽宁省自然科学基金面上项目(2023-MS-232)。
作者简介
第一作者:于丹(1983-),女,博士,副教授,主要从事中医调节肿瘤转移作用及其机制研究;通信作者:刘春英(1962-),女,教授,博士生导师,主要从事中医药治疗肿瘤机制研究。
