马鞭草总黄酮对HepG-2细胞增殖及侵袭力影响

Impact of total flavonoids of Verbena officinalis L on proliferation and invasiveness of human hepatocellular carcinoma HepG-2 cells

目的探讨马鞭草总黄酮(TFV)对肝癌HepG-2细胞增殖和侵袭力的影响。方法实验设对照组、TFV 50、100、200 mg/L组,噻唑蓝法检测人肝癌HepG-2细胞增殖率,Hoechest 33342染色检测HepG-2细胞凋亡情况,Transwell法检测HepG-2细胞侵袭力变化,ELISA法测定HepG-2细胞中基质金属蛋白酶9(MMP-9)及血管内皮生长因子(VEGF)表达水平。结果各剂量TFV均能明显抑制HepG-2细胞生长,呈现剂量依赖性(P<0.05);与对照组比较,TFV 50、100、200 mg/L组HepG-2细胞凋亡率[分别为(10.31±0.51)%、(16.02±0.63)%、(35.05±1.12)%]明显增加,透膜细胞数[分别为(119.2±17.1)、(98.5±13.5)、(54.8±6.4)个]明显下降,差异均有统计学意义(P<0.05);与对照组比较,TFV 50、100、200 mg/L组HepG-2细胞MMP-9和VEGF表达水平[分别为(7.81±0.48)、(6.28±0.41)(3.08±0.35)和(0.87±0.07)、(0.63±0.04)、(0.45±0.06)]明显降低,差异有统计学意义(P<0.05)。结论马鞭草总黄酮可明显抑制人肝癌HepG-2细胞增殖、降低HepG-2细胞侵袭力,其机制可能与下调HepG-2细胞内MMP-9和VEGF表达水平有关。

Objective To explore the effect of total flavonoids of Verbena officinalis L （TFV）on proliferation and invasiveness of human hepatocellular carcinoma （HepG-2） cells. Methods HepG-2 were treated with TFV at dosages of 50,100, and 200 mg/L ;the proliferation of HepG-2 cells was assayed by using 3-（4,5-dimethylthiazolyl）-2,5-diphenyl- tetrazolium bromide （MTT） method and the apoptosis of the cells was detected with Hoechest 33342 ;the invasiveness of the cells was detected with TransweU. The expressions of matrix metaUoproteinase-9 （MMP-9） and vascular endothelial growth factor （VEGF） were assessed with enzyme-linked immunosobent assay （ELISA）. Results Compared with the- control group, different concentration of TFV could inhibit the growth of HepG-2 cells significantly （ P 〈 0. 05 ） and sig- nificantly promote apoptosis of the cells, with the apoptosis rates of 10. 31 ± 0. 51%, 16. 02 ± 0. 63%, and 35.05 ± 1.12% and the invasiveness rates of 119. 2 ± 17.1,98. 5 ± 13.5 ,and 54. 8 ±6. 4 for the HepG-2 cells with the TFV dosa- ges of 50,100, and 200 mg/L, respectively （ all P 〈 0. 05 ）. The expression levels of MMP-9 （7.81 ± 0.48,6. 28 ± 0. 41, and 3.08 ±0. 35） and VEGF （0. 87 ±0.07,0. 63 ±0.04,and 0. 45 ±0. 06） were down-rugulated for the HepG-2 cells with the TFV dosages of 50,100, and 200 mg/L, respectively （ all P 〈 0. 05 ）. Conclusion Total flavonoids of Verbena officinalis L can inhibit the proliferation of HepG-2 cells and reduce invasiveness of the cells, and the mechanism of the effect may be related to down-regulation MMP-9 and VEGF.