hepaCAM与藏花素联合应用对前列腺癌细胞增殖的影响

Effects of hepaCAM combined with crocin on cell proliferation in prostate cancer cell line DU145

摘要:

目的：研究肝细胞黏附分子（hepatocyte cell adhesion molecule，hepaCAM）与藏花素联用对前列腺癌（prostate cancer，PCa）细胞DU145增殖的影响。方法：四甲基偶氮唑盐（methylthiazolyldiphenyl-tetrazolium bromide，MTT）法检测细胞增殖效应；克隆形成实验检测细胞克隆形成率；实时荧光定量PCR、Western blot法检测细胞周期调节因子cyclinD1及增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）的表达变化。结果：hepaCAM过表达腺病毒与藏花素单独使用均能抑制PCa细胞DU145的增殖[48 h抑制率分别为（12.0±2.0）%，（22.0±1.0）%]，但两者联用比单独应用效果更加明显[48 h抑制率为（54.0±1.0）%，P=0.000]。Real-time PCR结果显示，hepaCAM过表达腺病毒及藏花素联合作用与藏花素单独使用相比，cyclinD1 mRNA表达下调（P=0.048）；两者联合应用与hepaCAM过表达腺病毒或藏花素单独使用相比，PCNA mRNA水平下调（P=0.047，P=0.006）。Western blot结果显示，与单独使用hepaCAM过表达腺病毒或藏花素相比，两者联用组cyclinD1蛋白表达明显降低（P=0.002，P=0.002），同时PCNA蛋白表达也降低（P=0.000，P=0.003）。结论：hepaCAM基因过表达腺病毒与藏花素联用对PCa DU145细胞增殖的抑制有明显的协同作用，作用机制与下调cyclinD1和PCNA的表达有关。

Abstract:

Objective：To investigate the effects of hepatocyte cell adhesion molecule（hepaCAM） overexpression adenovirus combined with crocin on cell proliferation in prostate cancer cell line DU145. Methods：MTT assay was used to evaluate the effect of cell pro－liferation. Colony formation test was used to measure colony formation rate of single cell. Real-time quantitative PCR（qPCR） assay and Western blot were used to detect mRNA and protein expression of cyclinD1 and PCNA respectively. Results：After being treated with hepaCAM overexpression adenovirus or crocin alone，the proliferation of DU145 cell was inhibited[IR=（12.0±2.0）%，（22.0±1.0）%]，but the effect was more remarkable when the cells being treated with hepaCAM overexpression adenovirus and crocin combined[IR=（54.0±1.0）%，P=0.000]. Expression of cyclinD1 mRNA was down-regulated after being treated with hepaCAM over－expression adenovirus and crocin combined compared with being treated with crocin alone detected by qPCR（P=0.048），and prolif－erating cell nuclear antigen（PCNA） mRNA was down-regulated after being treated with hepaCAM overexpression adenovirus and crocin combined compared with being treated with hepaCAM overexpression adenovirus or crocin alone（P=0.047，P=0.006）. Western blot showed that compared with hepaCAM group or crocin group，expressin of cyclinD1 protein was down-regulated in hepaCAM and crocin combined group（P=0.002，P=0.002），meanwhile expression of PCNA protein was also down-regulated（P=0.000，P=0.003）. Conclusion：hepaCAM overexpression adenovirus combined with crocin can inhibit cell proliferation of prostate cancer cell line DU145 and the machanism is associated with down-regulation of cyclinD1 and PCNA expression.