吴茱萸碱对人肝癌HepG2细胞增殖影响

Impact of evodiamine on proliferation of human hepatocellular carcinoma Hep G2 cells

目的探讨吴茱萸碱(EVO)对人肝癌Hep G2细胞增殖的影响及作用机制。方法以人肝癌细胞株(Hep G2细胞)为受试细胞,实验设对照组、吴茱萸碱5.0、12.5、25.0、50.0、100.0μmol/L组,采用噻唑蓝法检测Hep G2细胞增殖抑制率,划痕实验检测Hep G2细胞伤痕愈合率,Western blot检测Hep G2细胞基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)蛋白。结果吴茱萸碱对Hep G2细胞增殖抑制率明显高于对照组,呈现时间-剂量依赖关系(P<0.05);与对照组比较,吴茱萸碱5.0、25.0、50.0μmol/L组Hep G2细胞24、48 h伤痕愈合率[分别为(23.31±1.03)%、(10.37±1.06)%、(7.92±1.40)%和(13.86±5.57)%、(1.78±3.08)%、0%]明显下降,差异有统计学意义(P<0.05);与对照组比较,吴茱萸碱5.0、25.0、50.0μmol/L组Hep G2细胞24 h MMP-9、PCNA蛋白表达量[分别为(0.53±0.020)、(0.35±0.020)、(0.21±0.015)与(0.76±0.032)、(0.66±0.01)、(0.59±0.015)]降低,差异有统计学意义(P<0.05)。结论吴茱萸碱可明显抑制Hep G2细胞增殖及远处迁徙,其机制可能与吴茱萸碱下调MMP-9、PCNA蛋白有关。

Objective To study the effect and mechanism of evodiamine on proliferation of human hepatocellular carcinoma（ HepG2） cells. Methods HepG2 cells were treated with evodiamin at dosages of 5. 0,12. 5,25. 0,50. 0,and100. 0 umol / L. The cell proliferation was assayed by using 3-（ 4,5-dimethylthiazolyl）-2,5-diphenyltetrazolium bromide（ MTT） method. The ability of cicatrization was analyzed with scratch wounds healing. The expressions of matrix metalloproteinase-9（ MMP-9）,proliferating cell nuclear antigen（ PCNA） in HepG2 cells were assessed with Western blot.Results The proliferation inhibition rate of evodiamine-treated groups was higher than that of the control group in a dose- and time-effect manner（ P〈0. 05）. Evodiamine could significantly inhibit cicatrization rate of HepG2 cells compared with the control group,with the cicatrization rates of 23. 31 ± 1. 03%,10. 37 ± 1. 06%,and 7. 92 ± 1. 40% at 24 hours and 13. 86 ± 5. 57%,1. 78 ± 3. 08,and 0 ± 0% at 48 hours for the groups with the dosage of 5. 0,25. 0,and 50. 0μmol /L,respectively（ all P〈0. 05）. The expression levels of MMP-9 and PCNA were down-regulated（ 0. 53 ± 0. 020,0. 35 ± 0. 020,and 0. 21 ± 0. 015 and 0. 76 ± 0. 032,0. 66 ± 0. 01,and 0. 59 ± 0. 015 for the groups with the dosage of5. 0,25. 0,and 50. 0 μmol / L）（ all P〈0. 05）. Conclusion Evodiamine can inhibit proliferation of HepG2 cells and the mechanism of the effect may be related to dow n-regulations of M M P-9 and PCNA proteins.