肿瘤相关巨噬细胞内Notch⁃1通过抑制组织蛋白酶S过表达影响非小细胞肺癌腺癌侵袭转移的机制研究  

Mechanism of Notch⁃1 in Tumor⁃associated Macrophages Affecting the In⁃vasion and Metastasis of Non⁃small Cell Lung Adenocarcinoma by Inhibi⁃ting the Overexpression of Cathepsin

作　　者：李海斌 吴振虎 丁建峰 LI Haibin;WU Zhenhu;DING Jianfeng(Department of Medical Imaging,Yuncheng Central Hospital of Shanxi Province,Yuncheng,Shanxi 044000,China)

机构地区：[1]山西省运城市中心医院医学影像科,山西运城044000

出　　处：《临床误诊误治》2024年第16期88-95,共8页Clinical Misdiagnosis & Mistherapy

基　　金：山西省医学重点科研项目计划(YJ2023001)。

摘　　要：目的探讨Notch-1在合并及未合并肝转移非小细胞肺癌腺癌患者肿瘤相关巨噬细胞内的表达情况,并通过体外细胞培养分析其对癌细胞侵袭转移的影响及相关机制。方法收集2021年1月至2022年12月收治的64例非小细胞肺癌腺癌患者标本,并分为未合并肝转移组32例和合并肝转移组32例,利用流式分选法分离CD68+肿瘤相关巨噬细胞,应用Western blot法检测2组CD68+肿瘤相关巨噬细胞Notch-1及相关蛋白表达水平。采用体外培养人单核/巨噬细胞THP-1细胞分别感染Notch-1 NC(阴性对照)和shRNA慢病毒载体后与人非小细胞肺癌A549共培养,应用Western blot法检测THP-1细胞相关蛋白表达情况,Transwell实验及划痕实验检测共培养体系中A549细胞迁移及侵袭能力。结果与未合并肝转移组比较,合并肝转移组CD68+肿瘤相关巨噬细胞内Notch-1表达水平显著降低,并伴随精氨酸酶-1及组织蛋白酶S表达水平显著增高(P<0.05)。与Notch-1 NC组比较,Notch-1 shRNA组THP-1细胞Notch-1表达水平显著降低,并伴随组织蛋白酶S表达水平升高(P<0.05)。与感染Notch-1 shRNA的THP-1细胞共培养后抑制表达Notch-1的A549细胞侵袭和迁移细胞数目明显增多,且细胞划痕距离明显减小(P<0.05)。结论肿瘤相关巨噬细胞内Notch-1蛋白表达具有抑制非小细胞肺癌腺癌细胞侵袭及转移的作用。Objective To explore the expression of Notch-1 in tumor-associated macrophages of non-small cell lung adenocarcinoma patients with and without liver metastasis,and to analyze its effect on cancer cell invasion and metastasis through in vitro cell culture and related mechanisms.Methods We collected specimens from 64 patients with non-small cell lung adenocarcinoma admitted from January 2021 to December 2022,and divided them into non-combined liver metastasis group(n=32)and combined liver metastasis group(n=32).Flow cytometry was performed to isolate CD68+tumor-associ-ated macrophages,and Western blot was used to detect the expression levels of Notch-1 in CD68+tumor-associated macropha-ges and related proteins in both groups.In vitro cultured human monocyte/macrophage THP-1 cells were infected with Notch-1 NC(negative control)and shRNA lentivirus vectors,respectively,and co-cultured with human non-small cell lung cancer A549.Western blot was used to detect the expression of THP-1 cell-related proteins,and Transwell assay and scratch assay were used to detect the migration and invasion ability of A549 cells in the co-culture system.Results Compared with the non-combined liver metastasis group,the expression level of Notch-1 in CD68+tumor-associated macrophages was significant-ly reduced in combined liver metastasis group,accompanied by a significant increase in the expression levels of arginase-1 and cathepsin S(P<0.05).Compared with the Notch-1 NC group,the Notch-1 shRNA group had significantly reduced expres-sion level of Notch-1 in THP-1 cells and increased protein expression level of cathepsin S(P<0.05).After co-culture with THP-1 cells infected with Notch-1 shRNA,the number of A549 cells expressing Notch-1 that inhibited invasion and migration significantly increased,and the distance of cell scratches significantly decreased(P<0.05).Conclusion The Notch-1 pro-tein in tumor-associated macrophages has a role in inhibiting invasion and metastasis of non-small cell lung adenocarcinoma.

关 键 词：非小细胞肺癌 腺癌 NOTCH-1 肿瘤相关巨噬细胞 精氨酸酶-1 组织蛋白酶S 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]