Dubois Louise, Stridsberg Mats, Kharaziha Pedram, Chioureas Dimitris, Meersman Niels, Panaretakis Theocharis, Ronquist K Göran
Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
Prostate. 2015 Jul 1;75(10):1063-73. doi: 10.1002/pros.22990. Epub 2015 Mar 17.
Prostasomes are nanosized extracellular vesicles exocytosed by prostate epithelial cells. They have been assigned many roles propitious to sperm in favor of fertilization. Prostatic cancer cells can also produce and secrete extracellular vesicles.
We assessed using ELISA, the surface expression of chromogranin proproteins on prostasomes and malignant extracellular vesicles of four different prostate cancer cell-lines, two hormone sensitive and two hormone refractory. We used a panel of chromogranin A and chromogranin B antibodies against peptides in-between hypothetical cleavage sites along the proproteins.
A diverging pattern of chromogranin peptides was apparent when comparing prostasomes and malignant extracellular vesicles indicating a phenotypical change. We also compared western blot patterns (prostasomes and malignant extracellular vesicles) for selected antibodies that displayed high absorbances in the ELISA. Western blot analyses revealed various cleavage patterns of those proproteins that were analyzed in prostasomes and extracellular vesicles.
Chromogranins are constituents of not only prostasomes but also of malignant prostate cell-derived extracellular vesicles with different amino acid sequences exposed at the membrane surface giving rise to a mosaic pattern. These findings may be of relevance for designing new assays for detection or even possible treatment of prostate cancers.
前列腺小体是前列腺上皮细胞胞吐产生的纳米级细胞外囊泡。它们具有许多有利于精子受精的作用。前列腺癌细胞也能产生和分泌细胞外囊泡。
我们使用酶联免疫吸附测定(ELISA)评估了嗜铬粒蛋白前体蛋白在来自四种不同前列腺癌细胞系(两种激素敏感型和两种激素难治型)的前列腺小体和恶性细胞外囊泡上的表面表达。我们使用了一组针对前体蛋白中假设切割位点之间肽段的嗜铬粒蛋白A和嗜铬粒蛋白B抗体。
在比较前列腺小体和恶性细胞外囊泡时,嗜铬粒蛋白肽呈现出不同的模式,表明存在表型变化。我们还比较了在ELISA中显示高吸光度的选定抗体的蛋白质印迹模式(前列腺小体和恶性细胞外囊泡)。蛋白质印迹分析揭示了在前列腺小体和细胞外囊泡中分析的那些前体蛋白的各种切割模式。
嗜铬粒蛋白不仅是前列腺小体的组成成分,也是恶性前列腺细胞衍生的细胞外囊泡的组成成分,其膜表面暴露有不同的氨基酸序列,形成一种镶嵌模式。这些发现可能与设计用于检测甚至可能治疗前列腺癌的新检测方法有关。