【摘要】 目的:研究雄激素应答元件陷阱DNA(AREdecoyDNA)对前列腺癌细胞LNCaP生长活性和凋亡的影响。方法:人工合成双链硫代AREdecoyDNA并提取LNCaP细胞核蛋白,应用电泳迁移率变动分析(electrophoreticmobilityshiftassay,EMSA)检测AREdecoyDNA与雄激素受体的特异结合。2μg/mlAREdecoyDNA转染LNCaP细胞,48h后相差显微镜观察细胞超微结构变化;MTT比色法检测细胞生长活性;DNALadder检测细胞凋亡;流式细胞技术(FCM)测定细胞凋亡率。结果:EMSA显示,AREdecoyDNA能特异与核蛋白中雄激素受体结合。LNCaP细胞转染AREdecoyDNA后,镜下可见部分细胞出现凋亡形态学的改变,有凋亡小体形成,细胞体外生长受到抑制,DNALadder可见明显梯形条带。转染后48h的凋亡率为22.5%。结论:AREdecoyDNA能竞争结合雄激素受体(androgenreceptor,AR),阻断AR的作用而诱导LNCaP细胞凋亡。更多还原

【Abstract】 Objective: To study the effect of ARE decoy DNA on proliferation and apoptosis of prostate cancer cell line LNCaP. Methods: Firstly, phosphorothioated ARE decoy DNA was synthesized and nucleoprotein was extracted from LNCaP cells. Electrophoretic mobility shift assay(EMSA) was used to detect the special combination of ARE decoy DNA with protein factor-androgen receptor. Secondly, ARE decoy DNA was transfected into LNCaP cells. The cell ultrastructure was observed under microscope and the cell growth was studied by MTT assay 48h later. Meanwhile, the cells were collected for determining apoptotic rate by flow cytometric(FCM) analysis and chromosome DNA Ladder. Results: EMSA showed that ARE decoy DNA could bind to androgen receptor specially. After transfection, the cell morphology changed a lot and the cell growth was inhibited. The apoptotic rate was 22.5% and DNA ladder could be observed significantly. Conclusions: ARE decoy DNA can induce the apoptosis of prostate cancer cell line LNCaP through binding the AR competitively. Decoy DNA strategy may provide a new method for therapy of prostate cancer.更多还原