雷公藤内酯醇对人LNCaP细胞雄激素受体表达的调控及其机制

发布时间：2018-01-07 20:10

本文关键词：雷公藤内酯醇对人LNCaP细胞雄激素受体表达的调控及其机制　出处：《药学学报》2015年10期 　论文类型：期刊论文

【摘要】：本文研究了雷公藤内酯醇(triptolide,TP)对人前列腺癌LNCa P细胞内雄激素受体(androgen receptor,AR)表达影响的作用机制。采用实时定量PCR和Western blot方法检测TP处理后LNCa P细胞中AR的m RNA和蛋白表达的变化,以及使用TP和NF-κB抑制剂共处理后LNCa P细胞AR蛋白表达的变化;采用RF克隆法构建系列p GL3-AR启动子报告基因载体并转染至LNCa P细胞,检测TP对LNCa P细胞AR转录的调节作用;采用Western blot检测可能起调节作用的上游蛋白。结果表明,TP处理LNCa P细胞48 h后,LNCa P细胞AR的m RNA和蛋白的表达随着药物浓度增加而下降,同时AR靶基因PART1及前列腺特异性抗原(prostate specific antigen,PSA)的表达也受到抑制;成功构建系列p GL3-AR启动子报告基因载体,测序结果表明,插入的目的序列正确且无碱基突变,转染LNCa P细胞后双荧光素酶报告基因检测系统证实其具有启动子活性;启动子报告基因实验结果表明,TP在转录水平下调AR的表达;与AR启动子活性相关的PI3K/AKT/NF-κB通路蛋白表达下调。说明TP可能通过PI3K/AKT/NF-κB通路下调人前列腺癌LNCa P细胞AR的转录活性。
[Abstract]:This paper studied the effects of triptolide (triptolide, TP) on human prostate cancer LNCa cells in P (androgen receptor AR, androgen receptor) expression mechanism. The influence of changes of TP treatment was detected by real-time quantitative PCR and Western blot LNCa P in AR cells after M RNA and protein expression, and the expression of LNCa P cell AR protein using TP and NF- were treated by B inhibitors; construction of series of P GL3-AR promoter reporter gene vector and transfected into LNCa P cells by RF cloning method, detection of TP regulation of LNCa P AR expression; using Western detection of blot may play an important role in the regulation of the upstream protein. The results showed that TP treatment of LNCa P cells after 48 h, the expression of M RNA and LNCa P protein in AR cells decreased with the increase of drug concentration, and the AR target gene PART1 and prostate specific antigen (prostate specific, antigen, PSA) expression is inhibited The successful construction of P series; GL3-AR promoter reporter gene vector and sequencing showed that the sequence ofinserted correct and no mutation, P cells transfected with LNCa luciferase reporter gene system confirmed the promoter activity of promoter reporter gene; the experimental results show that TP at the transcriptional level and down regulate the expression of AR; the activity of AR promoter PI3K/AKT/NF- B pathway related protein expression was down regulated. The transcriptional activity of TP through PI3K/AKT/NF- B pathway by LNCa human prostate cancer cell line AR P.

【作者单位】：扬州大学临床医学院泌尿外科;扬州大学临床医学院妇产科;扬州大学临床医学院;扬州大学医学院临床医学系;江苏省中西医结合老年病防治重点实验室;
【基金】：国家自然科学基金资助项目(81102460) 江苏省高校自然科学研究面上项目资助项目(14KJB310026)
【分类号】：R737.25
【正文快照】： 前列腺癌(prostate cancer,PCa)是男性最常见的恶性肿瘤之一,在美国男性癌症致死原因中居于第二位[1],在我国其发病率和死亡率呈明显上升态势[2]。雷公藤内酯醇(triptolide,TP)是中药雷公藤(Tripterygium wilfordii)的主要活性成分,在肿瘤治疗方面的活性近年来倍受关注,其前药

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