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首页> 外文期刊>Cellular & molecular biology letters. >ONCOGENE-DEPENDENT SURVIVAL OF HIGHLY TRANSFORMED CANCER CELLS UNDER CONDITIONS OF EXTREME CENTRIFUGAL FORCE - IMPLICATIONS FOR STUDIES ON EXTRACELLULAR VESICLES
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ONCOGENE-DEPENDENT SURVIVAL OF HIGHLY TRANSFORMED CANCER CELLS UNDER CONDITIONS OF EXTREME CENTRIFUGAL FORCE - IMPLICATIONS FOR STUDIES ON EXTRACELLULAR VESICLES

机译：极端离心力条件下高度转化的癌细胞的癌基因依赖性生存-对细胞外囊泡研究的意义

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Extracellular vesicles (EVs), including exosomes, are a subject of intense interest due to their emission by cancer cells and role in intercellular communication. Earlier reports suggested that oncogenes, such as RAS, MET or EGFR, drive cellular vesiculation. Interestingly, these oncogenes may also traffic between cells using the EV-mediated emission and uptake processes. One of the main tools in the analysis of EVs are ultracentrifugation protocols designed to efficiently separate parental cells from vesicles through a sequence of steps involving increasing g-force. Here we report that ultracentrifugationonly EV preparations from highly transformed cancer cells, driven by the overexpression of oncogenic H-ras (RAS-3) and v-src (SRC-3), may contain clonogenic cancer cells, while preparations of normal or less aggressive human cell lines are generally free from such contamination. Introduction of a filtration step eliminates clonogenic cells from the ultracentrifugate. The survival of RAS-3 and SRC-3 cells under extreme conditions of centrifugal force (110,000 g) is oncogene-induced, as EV preparations of their parental non-tumourigenic cell line (IEC-18) contain negligible numbers of clonogenic cells. Moreover, treatment of SRC-3 cells with the SRC inhibitor (PP2) markedly reduces the presence of such cells in the unfiltered ultracentrifugate. These observations enforce the notion that EV preparations require careful filtration steps, especially in the case of material produced by highly transformed cancer cell types. We also suggest that oncogenic transformation may render cells unexpectedly resistant to extreme physical forces, which may affect their biological properties in vivo.
机译：包括外泌体在内的细胞外囊泡（EVs）由于受到癌细胞的释放以及在细胞间通讯中的作用而受到广泛关注。较早的报道表明，癌基因，例如RAS，MET或EGFR，可驱动细胞的囊泡形成。有趣的是，这些致癌基因也可能利用EV介导的发射和摄取过程在细胞之间运输。电动汽车分析中的主要工具之一是超速离心协议，旨在通过一系列涉及增加g力的步骤有效地将亲代细胞与囊泡分离。在这里，我们报告说，在致癌性H-ras（RAS-3）和v-src（SRC-3）过表达的驱动下，高度转化的癌细胞进行的超速离心纯电动汽车可能包含克隆性癌细胞，而正常或攻击性较低的制剂人类细胞系通常没有这种污染。过滤步骤的引入消除了超速离心中的克隆细胞。 RAS-3和SRC-3细胞在极端离心力（110,000 g）的条件下的存活是由癌基因诱导的，因为其亲本非致瘤细胞系（IEC-18）的EV制剂中可克隆的细胞数量可忽略不计。此外，用SRC抑制剂（PP2）处理SRC-3细胞可显着减少未过滤的超速离心物中此类细胞的存在。这些观察结果证实了电动车制备需要仔细的过滤步骤的想法，尤其是在高度转化的癌细胞类型产生的材料的情况下。我们还建议致癌性转化可能使细胞出乎意料地抵抗极端的物理力，这可能会影响它们在体内的生物学特性。

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