缺氧诱导因子-1α诱导人前列腺癌细胞的上皮-间质转化。

Hypoxia-inducible factor-1alpha induces the epithelial-mesenchymal transition of human prostatecancer cells.

作者信息

Luo Yong, He Da-lin, Ning Liang, Shen Shu-lin, Li Lei, Li Xiang

机构信息

Department of Urology, First Hospital of Xi'an Jiaotong University, Xi'an 710061, China.

出版信息

Chin Med J (Engl). 2006 May 5;119(9):713-8.

PMID:16701010

Abstract

BACKGROUND

Hypoxia-inducible factor-1alpha (HIF-1alpha) is a transcriptional factor that could improve the stimulation of angiogenesis and the metabolic adaptation of tumor cells to hypoxia. A recent study showed that HIF-1alpha could induce colon cancer cells epithelial-mesenchymal transition (EMT). However, no evidence indicates a similar correlation in human prostate cancer cells. This study was designed to evaluate the effect of HIF-1alpha over-expression on the EMT in human prostate cancer cells.

METHODS

We selected the appropriate cell line for HIF-1alpha induction from those EMT negative prostate cell lines through vimentin gene detection by RT-PCR. As the result, LNCaP cell line is the best one for further experiment. LNCaP cells were transfected with recombinant plasmid pcDNA3.1 (-)/HIF-1alpha and pcDNA3.1 (-) control vector by Lipofectamine 2000 system. The positive cell colonies were confirmed by indirect immunofluorescence labeling. Then Transwell polycarbonate filter was used to analyze the invasive potency. The expression of EMT associated proteins, E-cadherin and vimentin, was detected by Western blotting.

RESULTS

Among four of the EMT negative cell lines, LNCaP was the only one expressed the vimentin gene but not the associated protein. The expression level of HIF-1alpha in LNCaP/HIF-1alpha was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. The cell numbers of LNCaP/HIF-1alpha that penetrated through the Transwell filter were higher than that of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells, the expression of vimentin was up-regulated in LNCaP/HIF-1alpha, whereas the expression of E-cadherin was down-regulated.

CONCLUSIONS

Over-expression of HIF-1alpha stimulates the invasion potency of human prostate carcinoma cells through EMT pathway. The expression of E-cadherin and vimentin, playing established roles in EMT, could be regulated by HIF-1alpha in human prostate cancer cell line.

摘要

背景

缺氧诱导因子-1α（HIF-1α）是一种转录因子，可增强血管生成刺激以及肿瘤细胞对缺氧的代谢适应。最近一项研究表明，HIF-1α可诱导结肠癌细胞发生上皮-间质转化（EMT）。然而，尚无证据表明在人前列腺癌细胞中存在类似的相关性。本研究旨在评估HIF-1α过表达对人前列腺癌细胞EMT的影响。

方法

我们通过RT-PCR检测波形蛋白基因，从EMT阴性前列腺细胞系中选择适合诱导HIF-1α的细胞系。结果显示，LNCaP细胞系是进一步实验的最佳选择。采用Lipofectamine 2000系统将重组质粒pcDNA3.1(-)/HIF-1α和pcDNA3.1(-)对照载体转染至LNCaP细胞。通过间接免疫荧光标记确认阳性细胞集落。然后使用Transwell聚碳酸酯滤膜分析侵袭能力。通过蛋白质免疫印迹法检测EMT相关蛋白E-钙黏蛋白和波形蛋白的表达。

结果

在四种EMT阴性细胞系中，LNCaP是唯一表达波形蛋白基因但不表达相关蛋白的细胞系。LNCaP/HIF-1α中HIF-1α的表达水平明显高于LNCaP/pcDNA3.1和LNCaP。穿过Transwell滤膜的LNCaP/HIF-1α细胞数量高于LNCaP/pcDNA3.1和LNCaP。与LNCaP/pcDNA3.1和LNCaP细胞相比，LNCaP/HIF-1α中波形蛋白的表达上调，而E-钙黏蛋白的表达下调。