JAK-STAT途径活化人前列腺癌雄激素受体的研究

Activation of androgen receptor mediated by JAK-STAT signal transduction pathway in human prestate cancer

目的 研究4种集落刺激因子(CSF):粒细胞集落刺激因子(G-CSF)、粒细胞-单核细胞集落刺激因子(GM-CSF)、白细胞介素-3(IL-3)和干细胞因子(SCF)对前列腺癌PC-3M细胞雄激素受体(AR)活化的途径。方法 将含人雄激素受体和雄激素受体报告基因氯霉素乙酰转移酶质粒转染至人前列腺癌细胞株PC-3M,集落刺激因子作用于转染了雄激素受体及其报告基因的细胞,对照组中同时加入终浓度为 50 mmol/L的 JAK-STAT途径持异性阻断剂 AG-490,检测外源性雄激素受体和报告基因的表达量。结果 PC-3M细胞能够表达外源性雄激素受体,10～100μg/L的G-CSF、GM-CSF、IL-3和SCF均能激活PC-3M细胞中外源性雄激素受体,检测到CAT的表达量在(0.58±0.16)μg/g到(3.80±0.89)μg/g之间,其表达量与培养基中的CSF浓度成正比;在终浓度 50 mmol/L AG-490的对照组中报告基因表达量显著下降或检测不到表达。结论 SCF能够激活PC-3M中外源性雄激素受体表达,JAK-STAT途径可能是人前列腺癌细胞雄激素受体旁路激活的机制之一。

Objective To study the activated effect of androgen receptor by colony-stimulating factors and their corresponding signal transduction pathway. Methods The plasmid pSG5-AR containing human androgen receptor and its reporter gene chloramphenicol acetyltransferase (ARE-CAT) were transfected into the human hormone-independent prostate cancer cell line PG-3M. The cell was cultured with 10-100μg/L colony-stimulaing factors, including G-CSF, GM--CSF, IL-3 and stem cell factor (SCF). The expression of exogenous androgen gene was detected by SP immunuohistochemistry. The re- porter gene was detected using CAT--ELISA kit. In the control group, AG--490, the special phosphory- lation inhibitor of JAK--STAT, in the final concentration of 50 mmol/L, was added in the culture to block the phosphorylation of STAT2 and STAT3. Results The exgenous androgern receptor gene was ex- pressed successfully in PC-3M. CAT expression could be detected [(0. 58±0. 16)μg/g to (3. 80±0. 89) μg/g] in cells stimulated with colony stimulating factors. 50 mmol/L AG-490 could depress the expres- sion of CAT greatly or thoroughly. Conclusion The four kinds of colony-stimulating factors, G-CSF, GMCSF, IL--3 and SCF, in the conecntration of 10-100μg/L can activate the esogenous androgen recep- tor in prostate cancer cell line PC-3M and JAK-STAT is confirmed to be the pathway to deliver their sig- nals.