摘要
目的研究粒细胞-巨噬细胞集落刺激因子（GM-CSF）激活人激素非依赖性前列腺癌PC-3M细胞雄激素受体及其可能的信号转导途径。方法将人雄激素受体和雄激素受体激活报告基因氯霉素乙酰转移酶（CAT）转染至人雄激素非依赖性前列腺癌细胞株PC-3M,不同浓度的GM- CSF以及特异性Erk1／2磷酸化阻断剂PD98059作用后检测雄激素受体报告基因CAT表达量的变化和促分裂素原活化蛋白激酶（MAPK）途径：Erk1／2分子磷酸化的改变。结果20～100 nmol／L的GM-CSF均能激活PC-3M细胞的外源性雄激素受体,CAT的相应表达量为（0．58±0．16）～（3．80±0．89）ng／mg,CAT表达量与培养基中的GM-CSF浓度成正相关;同时检测到PC-3M细胞中Erk1／2分子磷酸化程度与培养基中的GM-CSF浓度相关,在终浓度50 mmol／L的PD98059阻断Erk1／2分子磷酸化后CAT表达量显著下降。结论GM-CSF能独立激活前列腺癌PC-3M细胞中的雄激素受体,Erk1／2分子磷酸化可能是GM-CSF旁路激活人前列腺癌细胞雄激素受体的机制。
Objective To study the activation of androgen receptor by GM-CSF and its corresponding signal transduetion pathway in androgen-independent prostate cancer. Methods The plasmid pSGS-AR containing human androgen receptor and plasmid containing AR reporter gene chloramphenieol aeetyltransferase （ARE-CAT） were co-transfected into the human androgen-independent prostate cancer cell line PC-3M. The expression of exogenous androgen receptor gene was examined with S-P immunohistochemistry. The reporter gene in the transfected cells was detected using CAT- ELISA method after the cells were treated with 10-100 nmol/L GM-CSF for 5 to 30 min, and the phosphorylation of Erk1/2 was examined by western blot at the same time. As a control, PD98059, the special phosphorylation inhibitor of Erk1/2 was added in the culture to block the phosphorylation of the key molecule of MAPK pathway. Results The exogenous androgen receptor gene expressed successfully in PC-3M. CAT expression was detected in cells stimulated with 5-50 nmol/L GM-CSF. The intensity of Erk1/2 phosphorylation was correlated with the concentration of GM-CSF positively. 50 mmol/L PD98059 could inhibit the expression of CAT greatly. Conclusions GM-CSF, in the concentration of 10- 100 nmol/L could activate androgen receptor in androgen-independent prostate cancer cell. Erk1/2 phosphorylation plays an important role in this procedure. MAPK pathway may be a possible bypath activation of androgen receptor in androgen-independent prostate cancer.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2007年第5期342-345,共4页
Chinese Journal of Urology
基金
国家自然科学基金资助项目（39600146）
