文献类型：期刊文献

中文题名：半夏泻心汤含药血清对胃癌细胞外泌体诱导的腹膜间皮细胞FN1、LAMC1表达及胃癌细胞与腹膜间皮细胞黏附、侵袭的影响

英文题名：Effect of Banxia Xiexin Decoction Containing Serum on Adhesion and Invasion of Gastric Cancer Cells and Peritoneal Mesothelial Cells Induced by Exosomes Derived from Gastric Cancer Cells and the Expressions of FN1 and LAMC1

作者：丁富勇[1];刘喜平[2];陈启明[3];戴丽蓉[2];王庆苗[2];施丽娟[2];朱中博[2];何玲[2]

第一作者：丁富勇

机构：[1]甘肃中医药大学附属医院,甘肃兰州730050;[2]甘肃中医药大学,甘肃兰州730000;[3]兰州大学第一医院,甘肃兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2023

卷号：30

期号：9

起止页码：108

中文期刊名：中国中医药信息杂志

外文期刊名：Chinese Journal of Information on Traditional Chinese Medicine

收录：CSTPCD;;CSCD:【CSCD_E2023_2024】；

基金：国家自然科学基金(81860813、81860782);甘肃省自然科学基金(21JR7RA388)。

语种：中文

中文关键词：半夏泻心汤;胃癌细胞;腹膜间皮细胞;外泌体;黏附;侵袭

外文关键词：Banxia Xiexin Decoction;gastric cancer cells;peritoneal mesothelial cells;exosome;adhesion;invasion

摘要：目的观察半夏泻心汤含药血清对胃癌细胞来源外泌体诱导的腹膜间皮细胞纤维连接蛋白1(FN1)和层粘连蛋白γ1(LAMC1)表达及胃癌细胞与腹膜间皮细胞黏附、侵袭的影响,探讨其抑制胃癌腹膜转移的作用机制。方法制备半夏泻心汤含药血清,提取人胃癌细胞NCI-N87外泌体,采用透射电镜观察及Western blot进行鉴定。将人腹膜间皮细胞HMrSV5分为正常对照组、模型组、5-氟尿嘧啶(5-FU)组和半夏泻心汤低、中、高剂量组,每组3个复孔,以NCI-N87细胞外泌体诱导HMrSV5细胞,并分别用10%含药血清培养,ELISA检测HMrSV5细胞上清液FN1和LAMC1含量,PCR检测HMrSV5细胞FN1和LAMC1mRNA表达,荧光酶标仪检测NCI-N87细胞与HMrSV5细胞黏附情况,Transwell小室检测NCI-N87细胞向HMrSV5细胞侵袭情况。结果透射电镜观察NCI-N87细胞外泌体呈椭圆或碟状囊泡结构,粒径介于40~80 nm,外泌体标志蛋白CD9、CD63高表达,钙网蛋白低表达。与正常对照组比较,模型组HMrSV5细胞上清液FN1和LAMC1含量明显增加,HMrSV5细胞FN1和LAMC1 mRNA表达明显升高,NCI-N87细胞和HMrSV5细胞黏附率明显升高,NCI-N87细胞向HMrSV5细胞侵袭数量明显增加(P<0.01);与模型组比较,半夏泻心汤含药血清干预72 h后,HMrSV5细胞上清液FN1和LAMC1含量减少,HMrSV5细胞FN1和LAMC1 mRNA表达降低,NCI-N87细胞和HMrSV5细胞黏附率降低,NCI-N87细胞向HMrSV5细胞侵袭数量减少,其中半夏泻心汤中、高剂量组差异有统计学意义(P<0.05,P<0.01)。结论半夏泻心汤可降低胃癌细胞NCI-N87外泌体诱导的腹膜间皮细胞HMrSV5 FN1和LAMC1蛋白及mRNA表达,降低胃癌细胞与腹膜间皮细胞黏附和侵袭能力,可能是半夏泻心汤治疗胃癌腹膜转移的作用机制之一。
Objective To observe the effects of Banxia Xiexin Decoction containing serum on adhesion and invasion of gastric cancer cells and peritoneal mesothelial cells induced by exosomes derived from gastric cancer cells and the expressions of FN1 and LAMC1;To explore the mechanism of inhibiting peritoneal metastasis of gastric cancer.Methods Banxia Xiexin Decoction containing serum was prepared.The exosomes of human gastric cancer cells NCI-N87 were extracted and identified by transmission electron microscopy and Western blot.Human peritoneal mesothelial cells HMrSV5 were divided into normal control group,model group,5-fluorouracil(5-FU)group and Banxia Xiexin Decoction low-,medium-and high-dosage groups,with 3 pores in each group.HMrSV5 cells were induced by exosomes from NCI-N87 and cultured with 10%medicated serum respectively.The contents of FN1 and LAMC1 in supernatant of HMrSV5 cells was detected by ELISA,the expressions of FN and LAMC1 mRNA in HMrSV5 cells was detected by PCR,the adhesion of NCI-N87 cells and HMrSV5 cells was detected by fluorescence microplate reader,and the invasion of NCI-N87 cells into HMrSV5 cells was detected by Transwell chamber.Results Transmission electron microscope observation of gastric cancer cell-derived exosomes showed elliptical or dish-shaped vesicles,with a particle size ranging from 40 to 80 nm.Exosomal marker proteins CD9 and CD63 were highly expressed,and calreticulin was lowly expressed.Compared with the normal control group,the contents of FN1 and LAMC1 in supernatant of HMrSV5 cells in the model group increased significantly,the expressions of FN1 and LAMC1 mRNA in HMrSV5 cells increased significantly,the adhesion rate of NCI-N87 cells and HMrSV5 cells increased significantly,and the number of NCI-N87 cells invading HMrSV5 cells increased significantly(P<0.01).Compared with the model group,the contents of FN1 and LAMC1 in supernatant of HMrSV5 cells decreased after 72 h of intervention of Banxia Xiexin Decoction containing serum,the expressions of FN1 and LAMC1 mRNA in HMrSV5 cells decreased,the adhesion rate of NCI-N87 cells and HMrSV5 cells decreased,and the number of NCI-N87 cells invading HMrSV5 cells decreased.There was statistical significance in Banxia Xiexin Decoction medium-and high-dosage groups(P<0.05,P<0.01).Conclusion Banxia Xiexin Decoction can decrease the expressions of FN1 and LAMC1 protein and mRNA in peritoneal mesothelial cells HMrSV5 induced by exosomes derived from gastric cancer cells NCI-N87,and weaken the adhesion and invasion ability of gastric cancer cells and peritoneal mesothelial cells.This may be one of the mechanisms of Banxia Xiexin Decoction in the treatment of peritoneal metastasis of gastric cancer.