Division of Pharmaceutical Biosciences, Centre for Drug Research, Faculty of Pharmacy, University of Helsinki, Helsinki, 00014, Finland.
Krefting Research Center, Department of Internal Medicine and Clinical Nutrition, University of Gothenburg, Gothenburg, 40530, Sweden.
BMC Cancer. 2017 Feb 1;17(1):92. doi: 10.1186/s12885-017-3087-x.
Multiple types of extracellular vesicles (EVs), including microvesicles (MVs) and exosomes (EXOs), are released by all cells constituting part of the cellular EV secretome. The bioactive cargo of EVs can be shuffled between cells and consists of lipids, metabolites, proteins, and nucleic acids, including multiple RNA species from non-coding RNAs to messenger RNAs (mRNAs). In this study, we hypothesized that the mRNA cargo of EVs could differ based on the EV cellular origin and subpopulation analyzed.
We isolated MVs and EXOs from PC-3 and LNCaP prostate cancer cells by differential centrifugation and compared them to EVs derived from the benign PNT2 prostate cells. The relative mRNA levels of 84 prostate cancer-related genes were investigated and validated using quantitative reverse transcription PCR arrays.
Based on the mRNA abundance, MVs rather than EXOs were enriched in the analyzed transcripts, providing a snapshot of the tumor transcriptome. LNCaP MVs specifically contained significantly increased mRNA levels of NK3 Homeobox 1 (NKX3-1), transmembrane protease serine 2 (TMPRSS2), and tumor protein 53 (TP53) genes, whereas PC-3 MVs carried increased mRNA levels of several genes including, caveolin-2 (CAV2), glutathione S-transferase pi 1 (GSTP1), pescadillo ribosomal biogenesis factor 1 (PES1), calmodulin regulated spectrin associated protein 1 (CAMSAP1), zinc-finger protein 185 (ZNF185), and others compared to PNT2 MVs. Additionally, ETS variant 1 (ETV1) and fatty acid synthase (FASN) mRNAs identified in LNCaP- and PC-3- derived MVs highly correlated with prostate cancer progression.
Our study provides new understandings of the variability of the mRNA cargo of MVs and EXOs from different cell lines despite same cancer origin, which is essential to better understand the the proportion of the cell transcriptome that can be detected within EVs and to evaluate their role in disease diagnosis.
多种类型的细胞外囊泡（EVs），包括微囊泡（MVs）和外泌体（EXOs），由构成细胞 EV 分泌组的所有细胞释放。EV 的生物活性货物可以在细胞之间交换，包含脂质、代谢物、蛋白质和核酸，包括从非编码 RNA 到信使 RNA（mRNA）的多种 RNA 种类。在这项研究中，我们假设 EV 的 mRNA 货物可能因 EV 细胞来源和分析的亚群而不同。
我们通过差速离心从 PC-3 和 LNCaP 前列腺癌细胞中分离 MVs 和 EXOs，并将其与来自良性 PNT2 前列腺细胞的 EV 进行比较。使用定量逆转录 PCR 阵列研究并验证了 84 个前列腺癌相关基因的相对 mRNA 水平。
根据 mRNA 丰度，MVs 比 EXOs 更富集于分析的转录本，提供了肿瘤转录组的快照。LNCaP MVs 特异性地包含 NK3 同源盒 1（NKX3-1）、跨膜蛋白酶丝氨酸 2（TMPRSS2）和肿瘤蛋白 53（TP53）基因的 mRNA 水平显著增加，而 PC-3 MVs 则携带 Caveolin-2（CAV2）、谷胱甘肽 S-转移酶 pi 1（GSTP1）、pescadillo 核糖体生物发生因子 1（PES1）、钙调蛋白调节 spectrin 相关蛋白 1（CAMSAP1）、锌指蛋白 185（ZNF185）和其他基因的 mRNA 水平增加，与 PNT2 MVs 相比。此外，在 LNCaP 和 PC-3 衍生的 MVs 中鉴定的 ETS 变体 1（ETV1）和脂肪酸合酶（FASN）mRNA 与前列腺癌进展高度相关。
我们的研究提供了对不同细胞系来源的 MV 和 EXOs 的 mRNA 货物变异性的新认识，尽管它们具有相同的癌症起源，但这对于更好地理解 EV 中可检测到的细胞转录组的比例以及评估其在疾病诊断中的作用至关重要。
