人前列腺癌细胞中肝X受体特异性激动剂GW3965对核受体结合蛋白1的影响

Effect of GW3965 on NRBP1 expression in prostate cancer cells

目的探讨人前列腺癌细胞系DU145细胞中肝X受体(liver X receptor,LXR)特异性激动剂GW3965对核受体结合蛋白1(nuclear receptor binding protein 1,NRBP1)的表达调控作用。方法采用MTT法检测不同浓度GW3965(浓度分别为0.5、5、10μmol/L)作用24 h后对DU145细胞增殖的影响。在人前列腺癌DU145细胞中用LXR的特异性激动剂GW3965处理24 h后,经逆转录-聚合酶链反应(RT-PCR)检测LXR特异性靶基因三磷腺苷结合A1(ATP-bindingcassette A1,ABCA1)转录水平的表达情况;随后在转录水平和翻译水平检测NRBP1的表达。结果 LXR特异性激动剂GW3965各实验组对细胞的活性和增殖无影响(P>0.05)。GW3965作用于DU145细胞后,ABCA1转录水平呈剂量依赖性升高,表明LXR在DU145细胞中具有功能活性。LXR经过活化后在转录和翻译水平对NRBP1的抑制作用呈剂量依赖性降低。结论 LXR在DU145细胞中可抑制NRBP1的表达。

Objective To investigate the effect of liver X receptor（LXR） specific agonist GW3965 on the expression of nuclear receptor binding protein 1 （NRBPI） in DU145 cell line of prostate cancer. Methods The effect of various GW3965 concentrations （0.5,5,10 μmol/L） on the proliferation of DU145 cells was detected by the MTF test. DU 145 cells were treated with GW3965 for 24 h and then ATP-binding cassette A 1 （ABCA 1 ） mRNA was detected by RT-PCR;NRBP1 mRNA and protein were assayed by using RT-PCR and Western blot. Results GW3965 had no effect on the activity and proliferation of DU145 cells （P〉0.05）. The level of ABCA 1 mRNA was close-dependently increased, after treating DU 145 cells with different concentrations of GW3965 for 24 h respectively, which demonstrated that LXR had the functional activity in DU145. NRBP1 were down- regulated in both mRNA and protein level after LXR activation. Conclusion LXR could inhibit the expression of NRBP1 in DU145 cells.