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

雄激素依赖性增殖的分子机制：前列腺上皮细胞的合成和凋亡。

• 批准号: 11671573
• 负责人: KISHIMOTO Taketoshi
• 金额: $ 1.02万
• 依托单位: Osaka City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671573/
• 关键词: prostate; androgen receptor; decoy; MAPK; norepinephrine; P44; 42; アンドロゲン; α1レセブター; ARE; アポトーシス

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.

我们推测，雄激素反应元件（ARE）诱饵不仅可以抑制雄激素依赖性前列腺癌的增殖，而且可以抑制雄激素受体（AR）基因突变引起的雄激素非依赖性前列腺癌的增殖。我们合成了一个23-mer ARE诱饵和DNA-蛋白质相互作用的凝胶迁移率变动分析使用从LNCaP细胞制备的核提取物进行了检查。观察到ARE诱饵与LNCaP核蛋白（最可能是AR）的特异性结合。接下来，通过脂质体转染用ARE诱饵转染LNCaP细胞。孵育24小时后，通过DNA片段化检查细胞凋亡的诱导。有丝分裂原活化蛋白激酶（MAPK）在蛋白激酶级联反应中起重要作用，在调节细胞增殖和分化中起重要作用。在血管平滑肌细胞中，α1-肾上腺素能刺激通过激活ERK 1/2 MAPK增加DNA合成和细胞增殖。我们研究了去甲肾上腺素（NE）是否激活MAPK并刺激前列腺上皮细胞和非上皮细胞的增殖。NE（10 μ M<-6>和10 μ <-7>M）可显著激活间质细胞和平滑肌细胞中的ERK 1/2 MAPK，而在上皮细胞中不激活。JNK和p38未被激活。去甲肾上腺素可显著增加两种非上皮细胞对^3H-胸苷的摄取，而α1-肾上腺素能受体可抑制去甲肾上腺素的摄取。结果提示NE可刺激前列腺非上皮细胞增殖。