转化生长因子-β调节人前列腺癌细胞中白细胞介素-8表达的特征分析

Characterization of TGF-beta-regulated interleukin-8 expression in human prostate cancer cells.

作者信息

Lu Shan, Dong Zhongyun

机构信息

Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA.

出版信息

Prostate. 2006 Jun 15;66(9):996-1004. doi: 10.1002/pros.20424.

DOI:10.1002/pros.20424

PMID:16541418

Abstract

BACKGROUND

Interleukin (IL)-8 and transforming growth factor (TGF)-beta1 are overexpressed in advanced prostate cancer. The purpose of this study was to investigate TGF-beta1-regulated IL-8 expression in prostate cancer cells.

METHODS

TGF-beta receptor expression was evaluated by real-time reverse-transcription PCR (RT-PCR) and Western blotting. TGF-beta1-regulated IL-8 expression was determined by real-time RT-PCR, enzyme-linked immunoabsorbance assay (ELISA), nuclear run-on, and IL-8 promoter reporter assay.

RESULTS

PC-3MM2 cells expressed type I and type II TGF-beta receptors (TbetaRI and TbetaRII). LNCaP cells expressed significantly lower level of TbetaRII. Constitutive expression of IL-8 was detected in PC-3MM2 cells and LNCaP cells engineered with TbetaRII (LNCaP-TbetaRII). TGF-beta1 stimulated IL-8 expression in dose- and time-dependent manners, which was blocked by cycloheximide (CHX) and actinomycin D (ActD). The nuclear run-on and IL-8 luciferase reporter assays show that TGF-beta1 activated IL-8 gene transcription.

CONCLUSIONS

TGF-beta1 signaling regulates IL-8 expression in prostate cancer cells and may contribute to the overexpression of IL-8 in human prostate cancer.

摘要

背景

白细胞介素（IL）-8和转化生长因子（TGF）-β1在晚期前列腺癌中过表达。本研究旨在探讨TGF-β1对前列腺癌细胞中IL-8表达的调控作用。

方法

通过实时逆转录PCR（RT-PCR）和蛋白质印迹法评估TGF-β受体的表达。采用实时RT-PCR、酶联免疫吸附测定（ELISA）、细胞核转录分析和IL-8启动子报告基因测定法来确定TGF-β1对IL-8表达的调控。

结果

PC-3MM2细胞表达I型和II型TGF-β受体（TβRI和TβRII）。LNCaP细胞中TβRII的表达水平显著较低。在PC-3MM2细胞和转染了TβRII的LNCaP细胞（LNCaP-TβRII）中检测到IL-8的组成型表达。TGF-β1以剂量和时间依赖性方式刺激IL-8表达，这被放线菌酮（CHX）和放线菌素D（ActD）所阻断。细胞核转录分析和IL-8荧光素酶报告基因测定表明，TGF-β1激活了IL-8基因转录。