无雄激素条件下表皮生长因子诱导前列腺癌细胞系增殖及雄激素受体磷酸化

Proliferation and androgen receptor phosphorylation of prostate cancer cells induced by epidermal growth factor at the absence of androgen

目的探讨在无雄激素的条件下,表皮生长因子对前列腺癌细胞增殖及雄激素受体磷酸化的影响。方法以LNCaP及LAPC4 AR为研究对象,在无雄激素的条件下,EGF处理后,Western blot方法测定LNCaP及LAPC4 AR磷酸化状态;siRNA转染方法敲除Src基因或Ack1基因,观察对AR磷酸化的影响;CCK-8检测细胞增殖;定时定量RT-PCR检测前列腺特异抗原及人体激肽释放酶2 mRNA表达。结果 EGF通过Src激酶和另一未知激酶导致AR Tyr-534及AR Tyr-267特异位点磷酸化;相比未用EGF对照组,EGF能够促进前列腺癌细胞增殖(P<0.05),增加前列腺特异抗原(P<0.05)及人体激肽释放酶2 mRNA表达(P<0.05)。结论 EGF通过细胞内非受体酪氨酸激酶使AR特异位点磷酸化,诱导前列腺癌细胞增殖。

Objective To investigate the effect of epidermal growth factor on the proliferation and androgen receptor phosphorylation of prostate cancer cells in the absence of androgen. Methods LNCaP and LAPC4 AR were used for the study, after EGF treatment,Western blot was used to detect AR phosphorylation in LNCaP and LAPC4 cells; Src and Ackl genes with siRNA transfection were knocked down to observe the effect on androgen receptor phos phorylation; Cell Counting Kit-8 （CCK-8） was used to measure the cell proliferation; Quantitive RT-PCR was used to analyze the expression of prostate-specific antigen and human kallikrein-2 mRNA. Results EGF can induce AR specific site phosphorylation at Try-534 and Try-267 by Src kinase and another unknown kinase; Compared with untreated control, EGF enharced the proliferation of prostate cancer cells（ P 〈 O. 05 ） and increase the expression of prostate-specific antigen（P 〈0. 05） and human kallikrein-2（P 〈0. 05）. Conclusions EGF can induce proliferationand AR phosphorylation of prostate cancer by intracellular non-receptor tyrosine kinases