A doxycycline-inducible urokinase receptor (uPAR) upregulates uPAR activities including resistance to anoikis in human prostate cancer cell lines.
A doxycycline-inducible urokinase receptor (uPAR) upregulates uPAR activities including resistance to anoikis in human prostate cancer cell lines.
复制标题
强力霉素诱导的尿蛋白酶受体（UPAR）上调了UPAR活动，包括对人前列腺癌细胞系中对厌氧菌的抗性。
DOI：
10.1186/1476-4598-6-34
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发表时间：
2007-05-17
期刊：
影响因子：
37.3
通讯作者：
Sehgal, Inder
中科院分区：
文献类型：
--
作者：
Hasanuzzaman, Mohammad;Kutner, Robert;Agha-Mohammadi, Siamak;Reiser, Jakob;Sehgal, Inder
尿激酶受体（uPAR）介导多种细胞过程，包括涉及前列腺癌转移的几个事件。这些活性中的许多通过uPAR与其蛋白水解配体尿激酶（uPA）结合而启动或增强。本研究的目的是建立并检测一种能够在人前列腺癌细胞系中表达uPAR和DsRed荧光蛋白的诱导型慢病毒系统。将DsRed-uPAR融合构建体插入慢病毒载体中。用这种病毒和含有反向四环素反式激活因子（rt-TA）的病毒转导人前列腺癌细胞系导致稳定的转基因，其在用强力霉素刺激后以剂量响应方式诱导uPAR和DsRed蛋白。免疫印迹和免疫荧光研究表明，在非诱导的前列腺癌细胞系中没有可检测到的uPAR表达。与未诱导的细胞相比，具有诱导的uPAR的细胞表现出与基质底物玻连蛋白的细胞粘附增加，并且净细胞增殖增加。最后，诱导的表达uPARs的前列腺癌细胞在悬浮液中生长时，在很长一段时间内对失巢凋亡具有抵抗力。这种多西环素诱导的慢病毒系统在体外产生可滴定水平的生物活性uPAR。该工具可用于剖析前列腺癌细胞中uPAR诱导后的细胞事件。
The urokinase receptor (uPAR) mediates a diverse array of cellular processes including several events involved in prostate cancer metastasis. Many of these activities are initiated or enhanced by uPAR binding to its proteolytic ligand, urokinase (uPA). Our objective in this study was to generate and test an inducible lentiviral system capable of expressing uPAR and DsRed fluorescent protein in human prostate cancer cell lines. A DsRed-uPAR fusion construct was inserted into a lentiviral vector. Transduction of human prostate cancer cell lines with this virus and with a virus containing a reverse-tetracycline transactivator (rt-TA) resulted in a stable transgene which induced both uPAR and DsRed proteins in a dose-responsive fashion upon stimulation with doxycycline. Immunoblots and immunofluorescence studies indicated no detectable uPAR expression in non-induced prostate cancer cell lines. Cells with induced-uPAR demonstrated increased cellular adhesion to the matrix substrate vitronectin and increased net cell proliferation compared to uninduced cells. Finally, induced uPAR-expressing prostate cancer cells were resistant to anoikis over an extended time period when grown in suspension. This doxycycline-inducible lentivirus system produces titerable levels of biologically active uPAR in vitro. This tool can be used to dissect cellular events following induction of uPAR in prostate cancer cells.