基于PI3K/Akt信号通路探讨黄芪多糖对肺腺癌A549/DDP细胞移植瘤EMT进程的影响  

Effect of Astragalus Polysaccharide on EMT of A549/DDP Lung Adenocarcinoma Xenograft:An Exploration Based on PI3K/Akt Signaling Pathway

作　　者：张颖[1] 王淳[2] 于丹[2] 高原[2] 井欢[2] 王莹[2] 刘春英[2] ZHANG Ying;WANG Chun;YU Dan;GAO Yuan;JING Huan;WANG Ying;LIU Chun-ying(Jining Medical University,Jining 272000,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)

机构地区：[1]济宁医学院,山东济宁272000 [2]辽宁中医药大学,沈阳110032

出　　处：《中国实验方剂学杂志》2021年第23期51-58,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81973735,81774184)。

摘　　要：目的:探讨黄芪多糖(APS)对转化生长因子-β_(-1)(TGF-β_(-1))诱导的肺腺癌A549/DDP细胞移植瘤上皮间质转化(EMT)进程的影响及其潜在分子机制。方法:BALB/c裸鼠随机分为空载组(TGF-β_(-1)空载A549/DDP细胞),模型组,顺铂组,联合组(TGF-β_(-1)过表达A549/DDP细胞)。联合组灌胃APS(0.3 g·kg^(-1)·d^(-1))+腹腔注射顺铂(0.003 5 g·kg^(-1));顺铂组腹腔注射顺铂(0.003 5 g·kg^(-1))。药物干预后,处死裸鼠,取移植瘤和肺。称量瘤质量,计算抑瘤率;镜下计数肿瘤肺转移数;苏木素-伊红(HE)染色观察肿瘤病理和肿瘤组织肺转移情况;免疫组化,蛋白免疫印迹法(Western blot),实时荧光定量聚合酶链式反应(Real-time PCR)检测移植瘤中EMT分子标志物E-钙黏蛋白(E-cadherin),波形蛋白(Vimentin),α-平滑肌肌动蛋白(α-SMA)和磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)的蛋白和mRNA的表达。结果:与空载组比较,模型组瘤质量、肺转移结节数增加(P<0.05),肿瘤细胞连接稀疏,细胞长梭样,肺部有大面积肿瘤结节,E-cadherin蛋白和mRNA表达降低,Vimentin,α-SMA蛋白和mRNA表达升高,磷酸化(p)-PI3K,p-Akt蛋白表达升高(P<0.05)。与模型组、顺铂组比较,联合组瘤质量、肺转移结节数减少(P<0.05);肿瘤细胞连接紧密,细胞圆形或椭圆形,无明显肺转移,E-cadherin蛋白和mRNA表达升高(P<0.05),Vimentin,α-SMA蛋白和mRNA表达降低(P<0.05),p-PI3K,p-Akt蛋白表达降低(P<0.05)。各组间PI3K,Akt蛋白表达差异无统计学意义。结论:APS对肺腺癌A549/DDP细胞EMT有抑制作用,这一作用可能与其抑制活化的PI3K/Akt蛋白表达有关。Objective:To investigate the effect of Astragalus polysaccharide(APS)on transforming growth factor-β_(-1)(TGF-β_(-1))-induced epithelial mesenchymal transition (EMT) of A549/DDP lung adenocarcinoma xenograft and its potential molecular mechanism.Method:BALB/c nude mice were randomly divided into the non-loading group(A549/DDP cells not loaded with TGF-β_(-1)),model group,cisplatin group,and combined group(A549/DDP cells overexpressing TGF-β_(-1)).Mice in the combined group were treated with intragastric administration of APS(0.3 g·kg^(-1)·d^(-1))and intraperitoneal injection of cisplatin(0.003 5 g·kg^(-1)),while those in the cisplatin group only received intraperitoneal injection of cisplatin(0.003 5 g·kg^(-1)).After drug intervention,the nude mice were sacrificed and the xenograft and lung were harvested,followed by the weighing of tumor and the calculation of the inhibition rate.The number of tumors metastasizing to the lung was counted under the microscope.The pathological features of tumors and their metastasis to the lung tumor were observed by hematoxylin-eosin(HE)staining.The protein and mRNA expression levels of EMT molecular markers E-cadherin,Vimentin,α-smooth muscle actin(α-SMA),and phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)in the xenograft were detected by immunohistochemistry,Western blot,and Real-time polymerase chain reaction(Real-time PCR).Result:Compared with the non-loading group,the model group exhibited increased tumor weight and pulmonary metastatic nodules(P<0.05),sparse tumor cell junctions,long spindle cells,massive metastatic nodules in the lung,down-regulated E-cadherin protein and mRNA expression,and up-regulated Vimentin andα-SMA protein and mRNA expression and p-PI3K and p-Akt protein expression(P<0.05).Compared with the model group and cisplatin group,the combined group displayed decreased tumor weight and pulmonary metastatic nodules(P<0.05),tight tumor cell junctions,round or oval cells,no obvious lung metastasis,up-regulated E-cadherin protein and mRNA exp

关 键 词：黄芪多糖 上皮间质转化(EMT) 磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt) 肺腺癌 移植瘤 

分 类 号：R22[医药卫生—中医基础理论] R273[医药卫生—中医学]