扁蒴藤素抑制NCI-H1299细胞迁移侵袭的作用与机制研究  

Effect and underlying mechanism of pristimerin on migration and invasion of NCI-H1299 cells

作　　者：雷雪萍 钟怡行 邓秋狄 余细勇 LEI Xueping;ZHONG Yihang;DENG Qiudi;YU Xiyong(School of Pharmaceutical Sciences,Key Laboratory of Molecular Target&Clinical Pharmacology,Guangzhou Medical University,Guangzhou 511436,Guangdong,China;GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University,Guangzhou 511436,Guangdong,China)

机构地区：[1]广州医科大学药学院,广东省分子靶标与临床药理重点实验室,广东广州511436 [2]广州医科大学生命科学学院,广东广州511436

出　　处：《上海中医药杂志》2020年第12期79-83,96,共6页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金重点项目(81330007,U1601227);广东省重点实验项目(J19132004);广东省中医药局项目(20201208);广东省医学科学技术研究基金项目(A2020289)。

摘　　要：目的探讨扁蒴藤素对非小细胞肺癌NCI-H1299细胞迁移侵袭的抑制作用及其分子机制。方法采用不同浓度(0~80μmol/L)的扁蒴藤素处理NCI-H1299细胞,CCK-8法检测细胞增殖的情况,确定扁蒴藤素对NCI-H1299细胞的非毒剂量,并以此作为后续的药效与分子机制研究剂量。划痕实验评价扁蒴藤素对NCI-H1299细胞横向迁移能力的影响;transwell迁移与侵袭实验评价扁蒴藤素对NCI-H1299细胞纵向迁移能力与侵袭能力的影响;免疫印迹法与荧光定量PCR实验分别测定上皮-间充质相关标记物蛋白与mRNA的表达水平。结果扁蒴藤素浓度<2.5μmol/L时,对NCI-H1299细胞无明显的细胞毒性。与空白对照组比较,非毒剂量的扁蒴藤素(0.5μmol/L、1.0μmol/L和2.0μmol/L)可明显抑制NCI-H1299细胞的横向迁移以及纵向的迁移与侵袭能力(P<0.01)。扁蒴藤素处理后,上皮细胞标记物E-cadherin与ZO1的表达水平明显升高,间充质细胞标记物N-caherin与Vimentin的表达水平明显降低(P<0.01)。结论扁蒴藤素具有抑制NCI-H1299细胞迁移与侵袭的能力,且这一作用可能与其调控NCI-H1299细胞的上皮-间充质转化相关。Objective To evaluate the effect of pristimerin on the migration and invasion of non-small cell lung cancer(NSCLC)NCI-H1299 cells,and explore the underlying mechanism.Methods NCI-H1299 cells were treated with different concentrations of pristimerin(0~80μmol/L).The proliferation of NCI-H1299 cells was detected by cell counting kit(CCK-8)assay,and the non-toxic dose of pristimerin to NCI-H1299 cells was determined,which was used as the follow-up dose for the study of pharmacodynamics and molecular mechanism.The effect of pristimerin on the lateral migration ability of NCI-H1299 cells was evaluated by wound-healing assay;the effect of pristimerin on the longitudinal migration and invasion ability of NCI-H1299 cells was evaluated by transwell migration and invasion assay;the protein and mRNA expression levels of epithelial-mesenchymal related markers were measured by Western blot and real-time quantification PCR assay,respectively.Results When the concentration of pristimerin was less than 2.5μmol/L,it had no obvious cytotoxicity to NCI-H1299 cells.Compared with the blank control group,the non-toxic dose of pristimerin(0.5μmol/L,1.0μmol/L and 2.0μmol/L)could significantly inhibit the lateral migration and longitudinal migration and invasion of NCI-H1299 cells(P<0.01).After treatment with pristimerin,the expression levels of epithelial markers E-cadherin and ZO1 were significantly increased,while the expression levels of mesenchymal cell markers N-caherin and Vimentin were significantly decreased(P<0.01).Conclusion Pristimerin can inhibit the migration and invasion of NCI-H1299 cells,and this effect may be related to the regulation of epithelialmesenchymal transformation of NCI-H1299 cells.

关 键 词：扁蒴藤素 非小细胞肺癌 肿瘤细胞迁移 肿瘤细胞侵袭 上皮-间充质转化 

分 类 号：R285[医药卫生—中药学]