Molecular mechanism for androgen-dependent prol : feration and apoptosis of prostatic epithelial cells.

我们假设雄激素反应元件(ARE)诱饵不仅能抑制雄激素依赖型前列腺癌的增殖，还能抑制雄激素受体(AR)基因突变所致的雄激素非依赖性前列腺癌的增殖。我们合成了一个23聚体的ARE诱饵，并用LNCaP细胞的核提取液通过凝胶迁移率改变分析了DNA与蛋白质的相互作用。观察到ARE诱饵与LNCaP核蛋白的特异性结合，很可能是AR。然后，用脂质体转染法将ARE诱骗基因导入LNCaP细胞。孵育24小时后，通过DNA片段化检测细胞的凋亡诱导作用。ARE诱饵可能成为雄激素依赖和非雄激素依赖性前列腺癌的潜在治疗工具。丝裂原活化蛋白激酶(MAPK)在调节细胞增殖和分化的蛋白激酶级联反应中发挥重要作用。在血管平滑肌细胞中，ERK1-肾上腺素能刺激通过激活α1/2MAPK来增加DNA合成和细胞增殖。我们检测了去甲肾上腺素(NE)是否激活MAPK并刺激前列腺上皮和非上皮细胞的增殖。NE(10^lt；-6&gt；和10^lt；-7&gt；M)可显著激活基质细胞和平滑肌细胞的ERK1/2 MAPK，而对上皮细胞无明显激活作用。JNK和p38未被激活。NE可显著增加两种非上皮细胞对~(3 H)-胸腺嘧啶核苷的摄取，但可被α1肾上腺素受体抑制。上述结果提示，去甲肾上腺素可促进非上皮性前列腺细胞的增殖。

We hypothesize that androgen responsive element (ARE) decoy can inhibit the proliferation of not only androgen dependent prostatic cancer but also androgen independent one due to androgen receptor (AR) gene mutations. We synthesized a 23-mer ARE decoy and DNA-protein interactions were examined by gel mobility shift assay using nuclear extract prepared from LNCaP cells. Specific binding of ARE decoy to the LNCaP nuclear protein, most likely to be AR, was observed. Next, LNCaP cells were transfected with ARE decoy by lipofection. After 24h incubation, induction of apoptosis was examined by DNA fragmentation. ARE decoy may become a potential therapeutic tool for both androgen dependent and independent prostatic cancers.Mitogen-activated protein kinases (MAPK) function in protein kinase cascades that play critical roles in regulating cell proliferation and differentiation. In vascular smooth muscle cells, α1-adrenergic stimulation increases DNA synthesis and cell proliferation via activation of ERK1/2 MAPK. We examined whether norepinephrine (NE) activates MAPK and stimulates the proliferation of prostatic epithelial and non-epithelial cells. ERK1/2 MAPK was significantly activated by NE (10^<-6> and 10^<-7> M) in stromal cells and smooth muscle cells while not in epithelial cells. JNK and p38 were not activated. The uptake of ^3H-thymidine was significantly increased by NE in both non-epithelial cells, which was inhibited by α1-adrenoceptor. These results suggest that NE may stimulate the proliferation of non-epithelial prostatic cells.