雄激素受体反义RNA抑制前列腺癌细胞对激素的增殖反应

Proliferative response of human prostate cancer cell to the hormones inhibited by androgen receptor antisense RNA

目的 观察前列腺癌细胞LNCaP整合雄激素受体 (AR)反义RNA后对雄激素、雌激素和孕激素的增殖反应性变化。 方法 分别用含不同浓度雄激素、雌激素和孕激素的培养基培养前列腺癌细胞LNCaP、LNCaPas AR(基因组DNA中整合了AR反义RNA逆转录病毒载体的LNCaP细胞 )和LNCaPNeo(基因组DNA中整合了空逆转录病毒载体的LNCaP细胞 ) ,观察其生长情况 ,台盼蓝染色并细胞计数。 结果 在 1nmol/L合成雄激素R1881、10nmol/L 17β 雌二醇或 1nmol/L孕酮作用下 ,LNCa Pas AR细胞计数分别为 (2 .6± 0 .2 )× 10 5、(2 .6± 0 .2 )× 10 5、(2 .5± 0 .1)× 10 5/ml,与LNCaPNeo和LNCaP相比 ,生长显著受抑制 (P <0 .0 5 ) ,LNCaPNeo与LNCaP细胞增殖差异无显著性意义 (P >0 .0 5 )。显微镜下见LNCaP和LNCaPNeo细胞生长状态良好 ,而LNCaPas AR细胞稀疏且有较多细胞死亡。 结论 AR反义RNA可改变前列腺癌细胞LNCaP的雄激素非依赖性特征 ,可同时抑制其对雄激素、雌激素及孕激素的增殖反应。

Objective To investigate the effects of AR antisense RNA on growth of LNCaP cultured in medium containing varied concentrations of R1881,17β-estradiol and progesterone. Methods LNCaP cells transfected with AR antisense RNA retroviral vector pL-AR-SN were designed as LNCaP as-AR.LNCaP cells containing empty vector pLXSN were designed as LNCaP Neo.LNCaP and LNCaP Neo were taken as controls.In vitro cell growth assay,proliferative cells of LNCaP and transfected LNCaPs were stained with trypan blue and then counted when they were cultured with R1881(synthetic androgen),17β-estradiol and progesterone. Results Growth of LNCaP as-AR was inhibited significantly (P<0.05) compared with that of LNCaP and LNCaP Neo at 1 nmol/L R1881,10 nmol/L 17β-estradiol and 1 nmol/L progesterone,respectively.There was no significant difference in cell proliferation between LNCaP and LNCaP Neo(P>0.05).Microscopic observation showed that LNCaP and LNCaP Neo cells grew well,but only few LNCaP as-AR cells were alive and much more cells died. Conclusions AR antisense RNA retroviral vector pL-AR-SN can change androgen-independent characteristics of LNCaP cells,which may shed a novel insight into the treatment of androgen-independent prostate cancer.