Exosomal ITGA3 interferes with non-cancerous prostate cell functions and is increased in urine exosomes of metastatic prostate cancer patients.
Exosomal ITGA3 interferes with non-cancerous prostate cell functions and is increased in urine exosomes of metastatic prostate cancer patients.
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DOI：
10.3402/jev.v2i0.22097
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发表时间：
2013
影响因子：
16
通讯作者：
Jimenez CR
中科院分区：
文献类型：
--
作者：
Bijnsdorp IV;Geldof AA;Lavaei M;Piersma SR;van Moorselaar RJ;Jimenez CR
癌细胞能够改变周围非癌细胞的蛋白质表达和行为。由前列腺癌（PCa）细胞分泌的外泌体可能在癌症转移中具有功能性作用，并为蛋白质生物标志物提供了有希望的来源。本研究的目的是确定外泌体中哪些蛋白质可以影响非癌细胞，并确定我们是否可以使用尿液外泌体蛋白来识别高危PCa患者。通过超离心分离外来体。通过transwell（侵袭）测定研究迁移和侵袭。通过LC-MS/MS进行蛋白质组学，并通过Western印迹验证鉴定的蛋白质。通过FACS测量荧光标记的PKH 67-外来体的细胞摄取。基于LNCaP-和PC 3-外泌体的基于质谱的蛋白质组学的比较蛋白质谱分析，我们选择了参与迁移/侵袭的ITGA 3和ITGB 1进行进一步分析。外泌体ITGA 3的抑制几乎完全降低了非癌性前列腺上皮细胞（prEC）的迁移和侵袭。与LNCaP外泌体相比，PC 3-外泌体通过prEC的外泌体的细胞摄取更高。最后，与良性前列腺增生或PCa相比，ITGA 3和ITGB 1在转移性患者的尿外泌体中更丰富（p<0.05）。这些数据表明外泌体ITGA 3和ITGB 1可能在操纵非癌性周围细胞中起作用，并且尿外泌体中ITGA 3和ITGB 1的测量具有以非侵入性方式鉴定患有转移性PCa的患者的潜力。
Cancer cells are able to change the protein expression and behavior of non-cancerous surrounding cells. Exosomes, secreted by prostate cancer (PCa) cells, may have a functional role in cancer metastasis and present a promising source for protein biomarkers. The aim of the present study was to identify which proteins in exosomes can influence non-cancerous cells, and to determine whether we can use urine exosomal proteins to identify high-risk PCa patients. Exosomes were isolated by ultracentrifugation. Migration and invasion were studied by the transwell (invasion) assay. Proteomics was performed by LC-MS/MS and identified proteins were validated by Western blotting. Cellular uptake of fluorescent labeled PKH67-exosomes was measured by FACS. Based on comparative protein profiling by mass spectrometry-based proteomics of LNCaP- and PC3-exosomes, we selected ITGA3 and ITGB1, involved in migration/invasion, for further analyses. Inhibition of exosomal ITGA3 reduced the migration and invasion of non-cancerous prostate epithelial cells (prEC) almost completely. Cellular uptake of exosomes by prEC was higher with PC3-exosomes compared to LNCaP exosomes. Finally, ITGA3 and ITGB1 were more abundant in urine exosomes of metastatic patients (p<0.05), compared to benign prostate hyperplasia or PCa. These data indicate exosomal ITGA3 and ITGB1 may play a role in manipulating non-cancerous surrounding cells and that measurement of ITGA3 and ITGB1 in urine exosomes has the potential to identify patients with metastatic PCa in a non-invasive manner.