Cell-Free Nucleic Acids Circulating in the Plasma of Colorectal Cancer Patients Induce the Oncogenic Transformation of Susceptible Cultured Cells
Cell-Free Nucleic Acids Circulating in the Plasma of Colorectal Cancer Patients Induce the Oncogenic Transformation of Susceptible Cultured Cells
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DOI：
10.1158/0008-5472.can-09-3513
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发表时间：
2010-01-15
期刊：
影响因子：
11.2
通讯作者：
Garcia-Olmo, Damian
中科院分区：
文献类型：
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作者：
Garcia-Olmo, Dolores C.;Dominguez, Carolina;Garcia-Olmo, Damian
已经提出血浆中的无细胞核酸通过易感细胞对此类核酸的转染样摄取参与肿瘤发生和转移的发展。这种假定的现象暂时被称为“基因转移”。“在本研究中，我们研究了健康个体和结肠癌患者血浆对培养细胞的影响。NIH-3 T3细胞和人脂肪源性干细胞（hASC）的培养物中加入来自K-ras突变的结肠直肠肿瘤患者或健康受试者的血浆样品，采用两种不同的方案：直接将血浆加入标准平板中的培养物中，以及在血浆和细胞之间不接触的情况下加入，这些细胞被0.4 μ m孔的膜隔开。在血浆处理的hASC中，通过实时PCR检测不到K-ras突变序列。相反，在大多数等离子体处理的NIH-3 T3细胞（鼠细胞）的培养物中，发生了人DNA的转移，如通过检测人K-ras序列、p53序列和β-珠蛋白编码序列所证实的。此外，用结肠癌患者的血浆培养的NIH-3 T3细胞发生了致癌性转化，如非肥胖型糖尿病-重度联合免疫缺陷小鼠在注射此类细胞后发生癌所示。将血浆与培养细胞分离的膜的显微镜分析证实血浆中完全不存在细胞。我们只观察到非细胞颗粒，直径为
It has been proposed that cell-free nucleic acids in the plasma participate in tumorigenesis and the development of metastases via transfection-like uptake of such nucleic acids by susceptible cells. This putative phenomenon is tentatively referred to as "genometastasis." In the present study, we examined the effects on cultured cells of plasma from healthy individuals and from patients with colon cancer. Cultures of NIH-3T3 cells and human adipose-derived stem cells (hASC) were supplemented with samples of plasma from patients with K-ras-mutated colorectal tumors or from healthy subjects using two different protocols: direct addition of plasma to cultures in standard plates and addition in the absence of contact between plasma and cells, which were separated by a membrane with 0.4-mu m pores. In plasma-treated hASCs, no K-ras-mutated sequences were detected by real-time PCR. In contrast, in most cultures of plasma-treated NIH-3T3 cells (murine cells), the transfer of human DNA occurred, as verified by the detection of human K-ras sequences, p53 sequences, and beta-globin-encoding sequences. Moreover, NIH-3T3 cells that had been cultured with plasma from patients with colon cancer were oncogenically transformed, as shown by the development of carcinomas in nonobese diabetic-severe combined immunodeficient mice after the injection of such cells. Microscopic analysis of membranes that had separated plasma from cultured cells confirmed the complete absence of cells in the plasma. We only observed noncell particles, having diameters of