蛋白酪氨酸磷酸酶 SHP-1 在白细胞介素-6 调控前列腺癌细胞中的作用。

Role of the protein tyrosine phosphatase SHP-1 in Interleukin-6 regulation of prostate cancer cells.

机构信息

Department of Laboratory Medicin, Tumour Biology, Lund University, Malmö University Hospital, Malmö, Sweden.

出版信息

Prostate. 2010 Oct 1;70(14):1491-500. doi: 10.1002/pros.21184.

DOI:10.1002/pros.21184

PMID:20687222

Abstract

BACKGROUND

Interleukin-6 (IL-6) is a multifunctional cytokine that has been implicated in the modulation of growth and progression of prostate cancer. Decreased expression of the tyrosine phosphatase SHP-1, involved in regulation of cytokine and tyrosine kinase receptor signaling, has been shown to be associated with less favorable outcome among prostate cancer patients.

METHODS

Parental LNCaP cells and an LNCaP-IL6+ subline, derived from parental LNCaP cells by continuous culture of the cells in the presence of recombinant IL-6 were used in the study. Expression of STAT3, pSTAT3, ERK, pERK, AKT, pAKT, PTEN, and SHP-1 was analyzed by immunohistochemistry, Western blots, cDNA microarray, quantitative PCRs, and reverse transcriptase PCRs. Proliferation and apoptosis of transfected cells were analyzed by caspase3/7 assay and flow cytometry.

RESULTS

Phosphorylation of ERK and STAT3 was increased in the LNCaP-IL6+ subline compared with LNCaP cells, whereas pAKT was decreased. Overexpression and inhibition experiments with SHP-1 siRNA showed that SHP-1 reduced proliferation and increased apoptosis in both cell lines. Microarray analysis revealed 80 up-regulated and 87 down-regulated SHP-1-related genes in the LNCaP-IL6+ cell line compared with LNCaP cells.

CONCLUSIONS

SHP-1 suppresses growth and increases apoptosis in both LNCaP and LNCaP-IL6+ cells, which suggests that SHP-1 could be a therapeutic target in prostate cancer, even when there is an IL-6-related growth advantage.

摘要

背景

白细胞介素 6（IL-6）是一种多功能细胞因子，它参与调节前列腺癌的生长和进展。酪氨酸磷酸酶 SHP-1 的表达减少，该酶参与细胞因子和酪氨酸激酶受体信号的调节，已经表明与前列腺癌患者预后较差相关。

方法

本研究使用亲本 LNCaP 细胞和源自亲本 LNCaP 细胞的 LNCaP-IL6+亚系，该亚系通过在存在重组 IL-6 的情况下连续培养细胞而产生。通过免疫组织化学、Western blot、cDNA 微阵列、定量 PCR 和逆转录 PCR 分析 STAT3、pSTAT3、ERK、pERK、AKT、pAKT、PTEN 和 SHP-1 的表达。通过 caspase3/7 测定和流式细胞术分析转染细胞的增殖和凋亡。

结果

与 LNCaP 细胞相比，LNCaP-IL6+亚系中 ERK 和 STAT3 的磷酸化增加，而 pAKT 减少。SHP-1 的过表达和抑制实验表明，SHP-1 减少了这两种细胞系的增殖并增加了凋亡。微阵列分析显示，与 LNCaP 细胞相比，LNCaP-IL6+细胞系中有 80 个上调和 87 个下调的 SHP-1 相关基因。