Isolation by size of epithelial tumor cells -: A new method for the immunomorphological and molecular characterization of circulating tumor cells
Isolation by size of epithelial tumor cells -: A new method for the immunomorphological and molecular characterization of circulating tumor cells
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DOI：
10.1016/s0002-9440(10)64706-2
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发表时间：
2000-01-01
影响因子：
6
通讯作者：
Paterlini-Bréchot, P
中科院分区：
文献类型：
--
作者：
Vona, G;Sabile, A;Paterlini-Bréchot, P
我们开发了一种新的检测方法，ISET(按上皮性肿瘤细胞大小分离)，它允许对癌症患者循环中的肿瘤细胞进行计数、免疫形态和分子表征，使用小至1毫升的外周血液样本。使用这种方法，由于上皮性肿瘤细胞比外周血白细胞大，可以通过过滤分离出它们。ISET参数是使用添加了肿瘤细胞系(HepG2、Hep3B、MCF-7、HeLa和LNCaP)的外周血液定义的，ISET可以检测到单个微管插入的肿瘤细胞，添加到1 mI的血液中，我们还证明了荧光原位杂交可以用于对使用ISET收集的肿瘤细胞进行染色体分析，基于聚合酶链式反应的遗传分析可以应用于ISET分离的细胞，作为例子，我们证明在过滤和激光显微切割后的单个Hep3B细胞中存在纯合子的p53缺失。最后，我们提供了在接受肿瘤切除的肝细胞癌患者体内应用ISET的证据，但不包括逆转录聚合酶链式反应，允许分析细胞形态，肿瘤细胞计数，以及在手术中肿瘤微栓子扩散到外周血中的展示。总体而言，ISET构成了一种新的方法，应该会打开分子医学的新视角。
We have developed a new assay, ISET (isolation by size of epithelial tumor cells), which allows the counting and the immunomorphological and molecular characterization of circulating tumor cells in patients with carcinoma, using peripheral blood sample volumes as small as 1 ml. Using this assay, epithelial tumor cells can be isolated individually by filtration because of their larger size when compared to peripheral blood leukocytes. ISET parameters were defined using peripheral blood spiked with tumor cell lines (HepG2, Hep3B, MCF-7, HeLa, and LNCaP), ISET can detect a single, micropipetted tumor cell, added to 1 mi of blood, We also demonstrate that fluorescence in situ hybridization can be used to perform chromosomal analyses on tumor cells collected using ISET, Polymerase chain reaction-based genetic analyses can be applied to ISET-isolated cells, and, as an example, we demonstrate homozygous p53 deletion in single Hep3B cells after filtration and laser microdissection. Finally, we provide evidence for the in vivo feasibility of ISET in patients with hepatocellular carcinoma undergoing tumor resection, ISET, but not reverse transcriptase-polymerase chain reaction, allowed analysis of cell morphology, counting of tumor cells, and demonstration of tumor microemboli spread into peripheral blood during surgery. Overall, ISET constitutes a novel approach that should open new perspectives in molecular medicine.