摘要
目的:评价雷公藤红素对非小细胞肺癌NCI-H1299细胞的迁移与侵袭作用,并探讨其初步的作用机制。方法:采用CCK-8实验评价雷公藤红素对NCI-H1299细胞增殖活性的影响,获得非毒剂量;采用伤口愈合实验和Transwell迁移实验观察雷公藤红素对NCI-H1299细胞迁移能力的影响;采用Transwell侵袭实验检测雷公藤红素对NCI-H1299细胞侵袭能力的影响;蛋白免疫印迹实验与实时荧光定量PCR实验检测雷公藤红素对NCI-H1299细胞中间充质标记物N-Cadherin、Vimentin、Snail、Slug与上皮标记物E-Cadherin、ZO-1的表达的影响。结果:雷公藤红素可以呈剂量依赖性地抑制NCI-H1299细胞活性,在低于2.5μmol/L的剂量下对其增殖无明显影响;雷公藤红素在0.5μmol/L、1.0μmol/L、2.0μmol/L的剂量下可以抑制NCI-H1299细胞的迁移、侵袭能力;蛋白免疫印迹实验与实时荧光定量PCR实验结果表明,雷公藤红素能促进上皮标记物,抑制间充质标记物N-cadherin、Vimentin、Snail与Slug的表达,上调E-cadherin、ZO-1蛋白的表达。结论:雷公藤红素可以明显抑制NCI-H1299细胞的迁移与侵袭能力,且这一作用可能与其逆转NCI-H1299细胞的上皮-间充质转化(EMT)相关。
Objective:To investigate the effects of celastrol on the migration and invasion of non-small lung cancer NCI-H1299 cells and explore its preliminary mechanism.Methods:CCK-8 assay was used to detect the effect of celastrol on the proliferation of NCI-H1299cells,and non-toxic concentration was obtained. Wound healing assay and transwell migration assay were used to evaluate the effect of celastrol on the migration ability of NCI-H1299 cells. Transwell invasion assay was used to detect the effect of celastrol on the invasion ability of NCI-H1299 cells. Western blot and quantitative real-time PCR assays were conducted to detect the effect of celastrol on the expressions of mesenchymal markers N-Cadherin,Vimentin,Snail,Slug and epithelial markers E-Cadherin and ZO-1 in NCI-H1299 cells.Results:Celastrol inhibited the proliferation of NCI-H1299 cells in a dose-dependent manner,and had no significant effect on the proliferation at a concentration lower than 2.5 μmol/L. Celastrol inhibited the migration and invasion of NCI-H1299 cells at the concentration of 0.5 μmol/L,1.0 μmol/L and 2.0 μmol/L. The results of western blot and quantitative real time PCR assays showed that celastrol promoted epithelial markers,inhibited the expressions of mesenchymal markers N-cadherin,Vimentin,Snail and Slug,upregulated the expressions of E-cadherin and ZO-1 protein.Conclusion:Celastrol can significantly inhibit the migration and invasion ability of NCI-H1299 cells,and this effect may be related to its reversal of epithelial mesenchymal transition(EMT)of NCI-H1299 cells.KEYWOEDScelastrol;non-small cell lung cancer;migration;invasion;epithelial mesenchymal transition.
作者
林凯莉
邓秋狄
雷雪萍
LIN Kaili;DENG Qiudi;LEI Xueping(Department of Preventive Medicine,School of Public Health,Guangzhou Medical University,Guangzhou 511436,Guangdong,China;GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University,Guangzhou 511436,Guangdong,China;Key Laboratory of Molecular Target&Clinical Pharmacology,School of Pharmaceutical Sciences,Guangzhou Medical University,Guangzhou 511436,Guangdong,China)
出处
《上海中医药大学学报》
CAS
2022年第2期43-48,共6页
Academic Journal of Shanghai University of Traditional Chinese Medicine
基金
广东省中医药局科研项目(20201208)
广东省医学科学技术研究基金项目(A2020289)。
