[关键词]
[摘要]
目的 研究黄芩茎叶总黄酮(SBTF)对结肠癌HCT116细胞凋亡、迁移和侵袭的影响。方法 采用CCK8法检测SBTF(5、10、20、50、100、200、400、600 μg·mL-1)对HCT116细胞增殖的影响;Annexin V/PI双染法检测SBTF (80、120、160 μg·mL-1)对HCT116细胞凋亡的影响;JC-1染色法检测SBTF对细胞线粒体膜电位的影响;Transwell小室迁移和侵袭实验检测SBTF对细胞迁移和侵袭的影响;Western blotting法检测SBTF对细胞凋亡、迁移和侵袭相关蛋白表达的影响。结果 与对照组比较,SBTF可明显抑制HCT116细胞增殖,干预24、48 h后的半数抑制浓度(IC50)值分别为156.50、98.59 μg·mL-1;SBTF显著提高HCT116细胞凋亡率(P<0.05、0.01);SBTF明显促进HCT116细胞线粒体膜电位改变;SBTF显著降低HCT116细胞迁移和侵袭率(P<0.05、0.01);SBTF显著上调HCT116细胞的Bax/Bcl-2值和cleaved Caspase-3蛋白表达(P<0.05、0.01),显著下调MMP-2、MMP-9蛋白表达(P<0.05、0.01)。结论 SBTF可通过诱导HCT116细胞凋亡和抑制细胞迁移、侵袭发挥抗结肠癌作用。
[Key word]
[Abstract]
Objective To investigate the effects of total flavonoids in Scutellaria baicalensis stem-leaf (SBTF) on the apoptosis, migration and invasion process of colon cancer HCT116 cells. Methods The effects of SBTF (5, 10, 20, 50, 100, 200, 400 and 600 μg·mL-1) on HCT116 cells proliferation were detected by CCK8 assay. The effects of SBTF (80, 120, 160 μg·mL-1) on cell apoptosis were detected by Annexin V/PI double staining. The effects of SBTF on mitochondrial membrane potential (MMP) were detected by JC-1 staining. The effects of SBTF on cell migration and invasion were detected by Transwell migration and invasion test. The effects of SBTF on the expression of apoptosis, migration and invasion related proteins were detected by Western blotting. Results Compared with control group, SBTF could significantly inhibit the proliferation of HCT116 cells, and the IC50 values after 24 h and 48 h were 156.50 and 98.59 μg·mL-1. SBTF significantly enhanced apoptosis of HCT116 cells (P<0.05 and 0.01). SBTF could promote the change of MMP in HCT116 cells. SBTF effectively inhibited migration and invasion of HCT116 cells (P<0.05 and 0.01). SBTF significantly up-regulated the expressions of Cleaved Caspase-3 and the ratio of Bax/Bcl-2 in HCT116 cells (P<0.05 and 0.01), and significantly down-regulated the expressions of MMP-2 and MMP-9 in HCT116 cells (P<0.05 and 0.01). Conclusion SBTF could play an anti-colon cancer role by regulating the process of apoptosis, migration and invasion of HCT116 cells.
[中图分类号]
R285.5
[基金项目]
自贡市人工种植中药资源现状调查研究(CWKY-2018Z-03);江苏省科技成果转化专项项目(BA2018002)