| 摘要: |
| 目的研究TLR4在人前列腺癌PC3细胞中的表达意义及相关胞内信号机制。方法利用TLR4特异性配体脂多糖(LPS)刺激人前列腺癌PC3细胞,分别于LPS刺激0、2、6、12、24、48 h后收集细胞和上清,通过RT-PCR和蛋白质印迹方法检测TLR4在基因和蛋白水平的表达变化,通过RT-PCR方法检测胞内TGF-β、VEGF、IL-8、COX-2和MMP3的mRNA表达变化和ELISA方法检测上清VEGF、IL-8蛋白的表达变化；为了进一步研究相关信号通路,分别采用MAPK和NF-κB信号通路抑制剂预处理PC3细胞,然后利用同等浓度的LPS刺激,分别于4 h和24 h后收集上清,通过RT-PCR和ELISA方法重复上述细胞因子的检测。结果在LPS刺激后,人前列腺癌PC3细胞的TLR4功能性表达上调,引起胞内TGF-β、VEGF、IL-8、COX-2和MMP3的mRNA表达升高和上清中VEGF、IL-8的分泌增多(P<0.05)；进一步研究表明胞内p38 MAPK和NF-κB信号通路参与了此过程。结论TLR4信号通过p38 MAPK和NF-κB信号通路促进VEGF和IL-8的分泌。 |
| 关键词: 前列腺肿瘤 Toll样受体4 血管内皮生长因子受体 白细胞介素8 信号转导 |
| DOI：10.3724/SP.J.1008.2010.0697 |
| 投稿时间：2010-03-14修订日期：2010-05-28 |
| 基金项目:国家自然科学基金(30771984, 30972688). |
|
| LR4 signaling promotes secretion of VEGF/IL-8 in prostate cancer PC3 cells and related mechanism |
| GUO Dong, LIU Qiu-yan*,liu qiuyan |
| (Institute of Immunology, National Key Laboratory of Medical Immunology, Second Military Medical University, Shanghai 200433, China) |
| Abstract: |
| bjectiveTo study TLR4 expression in human prostate cancer PC3 cells and the related intracellular signaling mechanisms.MethodsHuman prostate cancer PC3 cells were stimulated with TLR4-specific ligand lipopolysaccharide(LPS), then the cells and supernatants were collected 0, 2, 6, 12, 24, and 48 hours after LPS stimulation.TLR4 mRNA and protein expression was examined by reverse transcription-PCR and Western blotting analysis, respectively.The mRNA expression of TGF-β, VEGF, IL-8, COX-2, and MMP3 was also measured by reverse transcription-PCR, and the levels of VEGF, IL-8 in the supernatants were examined by ELISA.To further study the related signaling pathway, MAPK and NF-κB signaling pathways were blocked by specific inhibitors in PC3 cells before LPS stimulation; the cells were collected after 4 hours and the supernatants were collected after 24 hours; and the above mentioned factors were examined by reverse transcription-PCR and ELISA again.ResultsTLR4 expression was up-regulated by LPS stimulation in human prostate cancer PC3 cells, which significantly increased mRNA expression of TGF-β, VEGF, IL-8, COX-2, and MMP3 and secretion of VEGF and IL-8 in the supernatants (P<0.05); further study showed that p38 MAPK and NF-κB signal pathways were involved in the process.ConclusionTLR4 signaling promotes VEGF and IL-8 secretion through p38 MAPK and NF-κB signal pathways. |
| Key words: prostatic neoplasms Toll-like receptor 4 vascular endothelial growth factor receptors interleukin-8 signal transduction |
