临床肝胆病杂志2025，Vol.41Issue(10)：2082-2092,11.DOI:10.12449/JCH251019
鸢尾黄素(TEC)对肝癌细胞活力、迁移和凋亡的影响及其机制
Effect of tectorigenin on the viability,migration,and apoptosis of hepatoma cells and its mechanism
摘要
Abstract
Objective To investigate the effect of blueberry-derived tectorigenin(TEC)on hepatocellular carcinoma cell lines HepG2 and Huh7 and its mechanism.Methods TEC was extracted from blueberries and purified,and a bioinformatics analysis was performed to identify potential target genes and signaling pathways.HepG2 and Huh7 cell lines were used and divided into 0,30,60,and 90 μg/mL groups according to the concentration of TEC.CCK-8 assay was used to measure cell viability;wound healing assay and Transwell assay were used to assess the migration ability of cells;flow cytometry was used to measure cell apoptosis rate;Western Blot was used to measure the protein expression levels of CCNB1,p53,MDM2,Bax,Bcl-2,and active-Caspase 3.Cell models with low CCNB1 expression(NC group,si-NC group,si-NC+TEC group,si-CCNB1 group,and si-CCNB1+TEC group)and CCNB1 overexpression(OE-NC group,OE-NC+TEC group,OE-CCNB1 group,and OE-CCNB1+TEC group)were established to validate the targets.A one-way analysis of variance or two factors analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.The Wilcoxon signed rank sum test was used to compare the expression levels of genes between cancer tissue and paracancerous tissue.Results In HepG2 and Huh7 cells under the same concentration of TEC,cell viability at 24 hours of TEC intervention was significantly lower than that at 12 and 48 hours(all P<0.05),and at 24 hours of intervention,the TEC 90 μg/mL group had a significantly lower cell viability than the other groups(all P<0.05).Therefore,TEC intervention for 24 hours at a concentration of 90 μg/mL was used for subsequent studies.Compared with the TEC 0 μg/mL group,the 30,60,and 90 μg/mL groups had significant reductions in the number of migrated cells and wound healing rate(all P<0.05),and compared with the NC group and si-NC group,the si-NC+TEC group and the si-CCNB1 group for HepG2 and Huh7 cells had significant reductions in the number of migrated cells and wound healing rate(all P<0.05).Compared with the NC group and si-NC group,the si-NC+TEC group and the si-CCNB1 group for HepG2 and Huh7 cells had a significant increase in cell apoptosis rate(all P<0.05).For HepG2 cells,compared with the 0 μg/mL group,the 30,60,and 90 μg/mL groups had significant reductions in the protein expression levels of CCNB1 and Bcl-2(all P<0.05),and the 60 and 90 μg/mL groups had significant increases in the protein expression levels of p53,Bax,and active-Caspase 3(all P<0.001)and a significant reduction in the protein expression level of MDM2(both P<0.05).For Huh7 cells,compared with the 0 μg/mL group,the 30,60,and 90 μg/mL groups had a significant reduction in the protein expression level of CCNB1(all P<0.01);the 60 and 90 μg/mL groups had significant increases in the protein expression levels of p53 and Bax and a significant reduction in the protein expression level of MDM2(all P<0.05);the 90 μg/mL group had a significant reduction in the protein expression level of Bcl-2 and a significant increase in the protein expression level of active-Caspase 3(both P<0.01).Compared with the si-NC group,the si-NC+TEC group and the si-CCNB1 group for HepG2 and Huh7 cells had significant reductions in the protein expression levels of CCNB1,MDM2,and Bcl-2 and significant increases in the protein expression levels of p53 and Bax(all P<0.05).Compared with the OE-NC group,the OE-NC+TEC group for HepG2 and Huh7 cells had significant reductions in the protein expression levels of CCNB1 and MDM2 and a significant increase in the protein expression level of p53(all P<0.05),while the OE-CCNB1 group had significant increases in the protein expression levels of CCNB1 and MDM2 and a significant reduction in the protein expression level of p53(all P<0.05),and there were no significant differences in the protein expression level of CCNB1,MDM2,and p53 between the OE-CCNB1 group and the OE-CCNB1+TEC group(all P>0.05).Conclusion TEC can inhibit the proliferation and migration of HepG2 and Huh7 cells and promote their apoptosis in vitro,possibly by downregulating the expression of CCNB1 and activating the p53 signaling pathway.关键词
花青苷类/肝肿瘤/Hep G2细胞/细胞周期蛋白B1Key words
Anthocyanins/Liver Neoplasms/Hep G2 Cells/Cyclin B1引用本文复制引用
唐文敏,程明亮,祝娟娟..鸢尾黄素(TEC)对肝癌细胞活力、迁移和凋亡的影响及其机制[J].临床肝胆病杂志,2025,41(10):2082-2092,11.基金项目
贵州省科技厅基础研究计划项目(黔科合基础-ZK[2022]一般452) （黔科合基础-ZK[2022]一般452）
国家自然科学基金地区基金(82160119,82360122) The Basic Research Program of Guizhou Provincial Science and Technology Department[Qiankehe Foundation-ZK(2022)General 452] （82160119,82360122）
National Natural Science Foundation of China Regional Funds(82160119,82360122) （82160119,82360122）
