Hypoxic enhancement of exosome release by breast cancer cells.
Hypoxic enhancement of exosome release by breast cancer cells.
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DOI：
10.1186/1471-2407-12-421
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发表时间：
2012-09-24
期刊：
影响因子：
3.8
通讯作者：
Gleadle JM
中科院分区：
文献类型：
--
作者：
King HW;Michael MZ;Gleadle JM
外体是由肿瘤细胞分泌的纳米囊泡，在肿瘤进展过程中发挥旁分泌信号的作用，包括肿瘤-间质相互作用、激活增殖途径和给予免疫抑制。缺氧是实体肿瘤的一个重要特征，它可能通过外切体介导的信号转导促进肿瘤的进展、血管生成和转移。乳腺癌细胞系在中度(1%O2)和重度(0.1%O2)低氧条件下培养。从条件培养液中分离外切体，通过纳米颗粒跟踪分析(NTA)和免疫印迹法检测外切体蛋白CD63的含量，以评估缺氧对外切体释放的影响。低氧外切体部分通过实时逆转录聚合酶链式反应检测miR-210，并归一化为外源性和内源性对照基因。统计学意义用学生t检验确定，P值为 <； 0.05被认为显著。用NTA和CD63免疫印迹法检测，三种不同的乳腺癌细胞系暴露于中度(1%O2)和重度(0.1%O2)低氧时，条件培养液中的外切体数量显著增加。低氧诱导因子(HIF)羟基酶抑制剂二羟甲基甘氨酸激活低氧信号，导致外切体释放显著增加。在低氧暴露前将HIF-1αsiRNA导入细胞，可阻止低氧促进外切体的释放。低氧调节的miR-210被发现在低氧外切体组分中水平升高。这些数据提供了证据，表明低氧促进乳腺癌细胞释放外切体，这种低氧反应可能是由HIF-1α介导的。鉴于肿瘤细胞来源的外切体在肿瘤进展中的一个新的作用，这对于理解低氧肿瘤的表型具有重要的意义，即低氧癌细胞可能会向其微环境中释放更多的外切体，以促进自身的生存和侵袭。
Exosomes are nanovesicles secreted by tumour cells which have roles in paracrine signalling during tumour progression, including tumour-stromal interactions, activation of proliferative pathways and bestowing immunosuppression. Hypoxia is an important feature of solid tumours which promotes tumour progression, angiogenesis and metastasis, potentially through exosome-mediated signalling. Breast cancer cell lines were cultured under either moderate (1% O2) or severe (0.1% O2) hypoxia. Exosomes were isolated from conditioned media and quantitated by nanoparticle tracking analysis (NTA) and immunoblotting for the exosomal protein CD63 in order to assess the impact of hypoxia on exosome release. Hypoxic exosome fractions were assayed for miR-210 by real-time reverse transcription polymerase chain reaction and normalised to exogenous and endogenous control genes. Statistical significance was determined using the Student T test with a P value of < 0.05 considered significant. Exposure of three different breast cancer cell lines to moderate (1% O2) and severe (0.1% O2) hypoxia resulted in significant increases in the number of exosomes present in the conditioned media as determined by NTA and CD63 immunoblotting. Activation of hypoxic signalling by dimethyloxalylglycine, a hypoxia-inducible factor (HIF) hydroxylase inhibitor, resulted in significant increase in exosome release. Transfection of cells with HIF-1α siRNA prior to hypoxic exposure prevented the enhancement of exosome release by hypoxia. The hypoxically regulated miR-210 was identified to be present at elevated levels in hypoxic exosome fractions. These data provide evidence that hypoxia promotes the release of exosomes by breast cancer cells, and that this hypoxic response may be mediated by HIF-1α. Given an emerging role for tumour cell-derived exosomes in tumour progression, this has significant implications for understanding the hypoxic tumour phenotype, whereby hypoxic cancer cells may release more exosomes into their microenvironment to promote their own survival and invasion.
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影响因子：
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作者：
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发表时间：
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影响因子：
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