摘要
目的:探讨金银花多酚粗对非小细胞肺癌NCI-H1299细胞恶性生物学行为的影响及分子机制。方法:将非小细胞肺癌NCI-H1299细胞分为NC组、紫杉醇组、金银花多酚粗提物低、中、高剂量组、miR-100-5p组、miR-NC组、anti-miR-100-5p+高剂量组、anti-miR-NC+高剂量组;四甲基偶氮唑盐比色法(MTT)检测细胞活性;Transwell检测细胞迁移和侵袭;蛋白质印迹法检测E-Cadherin、N-Cadherin、Vimentin和Wnt/β-catenin通路相关蛋白表达;实时荧光定量PCR(RT-qPCR)检测miR-100-5p表达水平。结果:不同浓度金银花多酚粗提物处理后,NCI-H1299细胞活性降低,细胞迁移和侵袭数量减少,E-Cadherin、miR-100-5p表达水平升高,N-Cadherin、Vimentin表达水平降低(P<0.05)。过表达miR-100-5p后,NCI-H1299细胞活性升高,细胞迁移[(226±20.03)个]和侵袭数量[(171±15.03)个]增加,E-Cadherin表达水平(0.34±0.03)降低,N-Cadherin(0.88±0.08)、Vimentin表达水平(0.95±0.09)升高(P<0.05)。下调miR-100-5p可逆转金银花多酚粗提物对NCI-H1299细胞增殖、迁移侵袭及Wnt/β-catenin通路的影响。结论:金银花多酚粗提物可能通过上调miR-100-5p表达,抑制Wnt/β-catenin通路活化,进而抑制非小细胞肺癌NCI-H1299细胞增殖、迁移和侵袭。
Objective:To investigate the impact of honeysuckle polyphenol crude extract on the malignant biological behavior of non-small cell lung cancer(NSCLC)NCI-H1299 cells and the molecular mechanisms involved.Methods:NCI-H1299 cells were divided into NC group,paclitaxel group,and low,medium,and high-dose groups of honeysuckle polyphenol crude extract.Additionally,miR-100-5p group,miR-NC group,anti-miR-100-5p+high-dose group,and anti-miR-NC+high-dose group were established.MTT assay was used to detect cell viability,Transwell assay for cell migration and invasion,Western blot for E-Cadherin,N-Cadherin,Vimentin,and Wnt/β-catenin pathway-related protein expression,and real-time fluorescence quantitative PCR(RT-qPCR)for miR-100-5p expression.Results:After treatment with different concentrations of honeysuckle polyphenol crude extract,NCI-H1299 cell viability decreased,and the number of cell migration and invasion decreased.E-Cadherin and miR-100-5p expression levels increased,while N-Cadherin and Vimentin expression levels decreased(P<0.05).Overexpression of miR-100-5p increased NCI-H1299 cell viability,cell migration[(226±20.03)cells],and invasion[(171±15.03)cells],while E-Cadherin expression level decreased(0.34±0.03),and N-Cadherin(0.88±0.08)and Vi-mentin expression levels increased(P<0.05).Downregulating miR-100-5p could reverse the effects of hon-eysuckle polyphenol crude extract on NCI-H1299 cell proliferation,migration,invasion,and the Wnt/β-catenin pathway.Conclusion:Honeysuckle polyphenol crude extract may inhibit the proliferation,migration,and invasion of NSCLC NCI-H1299 cells by upregulating miR-100-5p expression,thereby suppressing the activation of the Wnt/β-catenin signaling pathway.
作者
申兴勇
白爽
李新涛
李亚红
王孝彬
SHEN Xingyong;BAI Shuang(Xi'an People's Hospital/Aerospace District of the Forth Hospital,Shaanxi Xi'an 710004,China)
出处
《河北医学》
2024年第1期23-28,共6页
Hebei Medicine
基金
陕西省重点研发计划项目,(编号:2022SF-230)。
