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膀胱癌细胞来源的外泌体抑制肿瘤细胞凋亡并诱导细胞增殖体外。

Bladder cancer cell-derived exosomes inhibit tumor cell apoptosis and induce cell proliferation in vitro.

机构信息

Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Yuzhong, Chongqing 400016, P.R. China.

出版信息

Mol Med Rep. 2013 Oct;8(4):1272-8. doi: 10.3892/mmr.2013.1634. Epub 2013 Aug 14.

DOI:10.3892/mmr.2013.1634
PMID:23969721
Abstract

Exosomes are small membrane vesicles released by a variety of mammalian cells into the extracellular space and are involved in cell‑to‑cell signaling. This study aimed to investigate the effects of bladder cancer cell‑derived exosomes on the regulation of tumor cell viability and apoptosis, as well as the underlying molecular events. Exosomes were purified from the supernatants of human bladder cancer T24 cell cultures. Transmission electron microscopy was used to confirm their morphology and western blot analyses determined the protein content of cells. Subsequently, bladder cancer cell lines were treated with different concentrations of exosomes. Tumor cell viability was shown to be reduced, as detected by the Cell Counting Kit‑8 assay. Annexin V/flow cytometric assays showed that exosomes inhibited apoptosis of bladder cancer cell lines in a dose- and time‑dependent manner. Exosomes were demonstrated to upregulate the expression of Bcl‑2 and Cyclin D1 proteins, but reduce the levels of Bax and caspase‑3 proteins in these cells. Moreover, exosomes dose‑dependently increased the expression of phosphorylated Akt and extracellular signal‑regulated protein kinase (ERK). In conclusion, this study demonstrated that bladder cancer cell‑derived exosomes inhibited tumor cell apoptosis, which was associated with the activation of Akt and ERK pathway genes, suggesting that tumor‑derived exosomes are involved in bladder cancer progression. Inhibition of exosome formation and release may therefore be a novel strategy in future treatment of bladder cancer.

摘要

外泌体是各种哺乳动物细胞释放到细胞外空间的小膜囊泡,参与细胞间信号传递。本研究旨在探讨膀胱癌细胞衍生的外泌体对肿瘤细胞活力和凋亡的调节作用及其潜在的分子事件。外泌体从人膀胱癌 T24 细胞培养物的上清液中纯化。透射电子显微镜用于确认其形态,Western blot 分析确定细胞的蛋白含量。随后,用不同浓度的外泌体处理膀胱癌细胞系。细胞计数试剂盒-8 检测显示肿瘤细胞活力降低。Annexin V/流式细胞术检测表明,外泌体以剂量和时间依赖的方式抑制膀胱癌细胞系的凋亡。外泌体被证明上调 Bcl-2 和 Cyclin D1 蛋白的表达,但降低这些细胞中 Bax 和 caspase-3 蛋白的水平。此外,外泌体还可剂量依赖性地增加磷酸化 Akt 和细胞外信号调节蛋白激酶 (ERK) 的表达。总之,本研究表明膀胱癌细胞衍生的外泌体抑制肿瘤细胞凋亡,这与 Akt 和 ERK 通路基因的激活有关,提示肿瘤衍生的外泌体参与膀胱癌的进展。因此,抑制外泌体的形成和释放可能是未来膀胱癌治疗的一种新策略。

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