参芪扶正注射液对肺腺癌A549/DDP细胞荷瘤裸鼠移植瘤KRT19/β-catenin信号通路的影响

Effect of Shenqi Fuzheng Injection(参芪扶正注射液)on KRT19/β-catenin Signal Pathway in Transplantation Tumor of Lung Adenocarcinoma with A549/DDP Cells

目的探讨参芪扶正注射液对肺腺癌A549/DDP细胞荷瘤裸鼠移植瘤角蛋白19(keratin 19,KRT19)/β-cate-nin信号通路的影响。方法40只BALB/c-nu裸鼠随机分为对照组,顺铂组,参芪扶正注射液组,顺铂联合参芪扶正注射液低剂量组,顺铂联合参芪扶正注射液高剂量组。每只裸鼠分别腋下接种A549/DDP细胞,第7天成瘤,第8~25天开始给予对应药物处理,第26天将全部裸鼠处死。检测各组瘤重和抑瘤率;HE染色观察各组肿瘤组织病理形态学改变;透射电镜观察各组肿瘤细胞超微结构变化;Western blot和免疫组化法检测移植瘤组织KRT19/β-catenin信号通路相关蛋白的表达。结果与顺铂组相比,顺铂联合参芪扶正注射液低、高剂量组瘤重降低(P<0.05),且随着参芪扶正注射液给药剂量增加,瘤重逐渐减小,抑瘤率随之增加;HE染色显示,与顺铂组相比,联合用药组核皱缩加深,核分裂较少,坏死灶增加;透射电镜结果显示,与顺铂组相比,联合用药组出现更多的固缩核、膜泡、内质网肿胀、线粒体变性、脂滴和微绒毛丢失等;与顺铂组相比,顺铂联合不同剂量参芪扶正注射液组可显著下调移植瘤组织KRT19、β-catenin、CD44的表达,上调E-cadherin的表达。结论参芪扶正注射液可通过KRT19/β-catenin信号通路增强A549/DDP荷瘤裸鼠移植瘤对顺铂的敏感性。

Objective To investigate the effects of Shenqi Fuzheng Injection(参芪扶正注射液)on the keratin 19(KRT19)/β-catenin signaling pathway in lung adenocarcinoma A549/DDP cell xenografts in nude mice.Methods Forty BALB/c-nu nude mice were randomly divided into the control group,cisplatin group,Shenqi Fuzheng Injection group,cisplatin combined with Shenqi Fuzheng Injection low dose group,and cisplatin combined with Shenqi Fuzheng Injection high dose group.A549/DDP cells were inoculated into axilla of each nude mouse respectively,and the tumor was formed on the 7th day.The corresponding drug treatment was started on the 8th-25th day,and all nude mice were executed on the 26th day.The tumor weight and tumor inhibition rate were measured in each group.HE staining was used to observe the pathomorphological changes of tumor tissues in each group.The ultrastructural changes of tumor cells in each group were observed by transmission electron microscope.The expressions of KRT19/β-catenin signaling pathway related proteins in the xenografts tissue were detected by Western blot and immunohistochemistry.Results Compared with those of the cisplatin group,the tumor weights of the cisplatin combined with Shenqi Fuzheng Injection low and high groups were decreased(P<0.05).With the increasing dose of Shenqi Fuzheng Injection,the tumor weights were gradually decreased,and the tumor inhibition rate was increased.HE staining showed that compared with the cisplatin group,the combination group had deeper nuclear shrinkage,less nuclear division and increased necrotic lesions.The results of transmission electron microscopy showed that compared with those of the cisplatin group,more pyknosis,vesicles,endoplasmic reticulum swelling,mitochondrial degeneration,lipid droplets and microvilli loss occurred in the combination group.Compared with the cisplatin group,the cisplatin combined with different doses of Shenqi Fuzheng Injection groups could significantly down-regulate the expressions of KRT19,β-catenin and CD44 in the xenografts tissue and up-regulate the expression of E-Cadherin.Conclusion Shenqi Fuzheng Injection can enhance the sensitivity of A549/DDP xenografts to cisplatin through the KRT19/β-catenin signaling pathway.