β-羟基异戊酰紫草素对前列腺癌细胞株的生长抑制作用及机制研究

Growth inhibitory effect of β-hydroxyisovalerylshikonin on prostate cancer cells and related mechanism

目的研究β-羟基异戊酰紫草素(β-HIVS)对激素非依赖性前列腺癌细胞株PC-3的体外生长抑制作用及其可能机制。方法采用MTT法检测β-HIVS对PC-3和人皮肤成纤维细胞株HSF的细胞生长抑制率;荧光素酶报告基因法检测给药后细胞中低氧诱导因子-1(HIF-1)和核因子κB(NF-κB)的转录活性;RT-PCR检测HIF-1下游靶基因血管内皮生长因子(VEGF)的表达;Westernblotting检测β-HIVS作用后HIF-1α蛋白的表达水平。结果β-HIVS对PC-3的细胞生长具有显著抑制作用(P<0.05),而对HSF的细胞生长无明显影响(P>0.05)。给药后PC-3细胞中NF-κB转录活性无明显变化(P>0.05);而HIF-1转录活性受抑(P<0.05),其下游靶基因VEGF表达下调,且细胞中HIF-1α蛋白累积减少。结论β-HIVS对PC-3的细胞生长具有抑制作用,其机制可能与β-HIVS减少低氧条件下PC-3中HIF-1α蛋白水平,抑制其转录活性,使下游促细胞生长靶基因VEGF表达下调有关。

Objective To investigate the growth inhibitory effects of β-hydroxyisovalerylshikonin（β-HIVS）on hormone-independent prostate cancer cell line PC-3,and explore the related mechanism.Methods The inhibition rates of β-HIVS on PC-3 cells and normal human skin fibroblasts（HSF）were determined by MTT method.Luciferase reporter assay was adopted to examine the transcriptional activity of hypoxia-inducible factor-1（HIF-1）and nuclear factor-kappa B（NF-κB）.The expression of vascular endothelial growth factor（VEGF）,the target gene of HIF-1,was detected by RT-PCR.The expression of HIF-1α protein in PC-3 cells treated by β-HIVS was detected by Western blotting.Results β-HIVS significantly inhibited the growth of PC-3 cells（P0.05）,while had no significant effect on HSF cells（P0.05）.There was no significant change in transcriptional activity of NF-κB in PC-3 cells after drug treatment（P0.05）.β-HIVS inhibited the transcriptional activity of HIF-1 and decreased the expression of VEGF.The protein level of HIF-1α in PC-3 cells was reduced by β-HIVS.Conclusion β-HIVS inhibits the growth of PC-3 cells by decreasing the level of HIF-1α protein in PC-3 cells under hypoxia condition,thus attenuating the transcriptional activity of HIF-1α and decreasing the expression of VEGF.