Multigene reverse transcription-PCR profiling of circulating tumor cells in hormone-refractory prostate cancer
Multigene reverse transcription-PCR profiling of circulating tumor cells in hormone-refractory prostate cancer
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DOI：
10.1373/clinchem.2003.028563
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发表时间：
2004-05-01
影响因子：
9.3
通讯作者：
Terstappen, LWMM
中科院分区：
文献类型：
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作者：
O'Hara, SM;Moreno, JG;Terstappen, LWMM
背景：循环肿瘤细胞（ctc）代表了组织的替代来源，在概念上代表了“实时”活检。我们之前报道过ctc的数量反映了激素难治性前列腺癌（HRPC）的疾病进展。为了进一步完善CTCs的表征，我们进一步研究了HRPC中CTCs是否可以获得基于体外转录的多基因逆转录pcr表达谱。方法：我们从健康供者和HRPC患者的7.5 ml血液样本中提取免疫磁富集的ctc，构建反义RNA文库，评估了37个具有上皮细胞表征潜力的基因的表达。结果：对照组37个基因中13个无表达。9例转移性前列腺癌患者23份血液标本ctc中表达最显著的基因为前列腺特异性抗原（20 / 23,87%）、前列腺特异性膜抗原（17 / 23,74%）、雄激素受体（16 / 23,70%）、人腺体钾化因子2（7 / 23,30%）、表皮生长因子受体（4 / 23,17%）和与G蛋白受体同源的前列腺特异性基因（2 / 23,9%）。这些样本中ctc的数量在7.5 mL血液中从4到283不等（平均87个，中位数89个）。一些基因在对照样本中表达较低，而在患者样本中表达较高。在所有23个样本中，细胞角蛋白19、上皮细胞粘附分子或粘蛋白1均有表达。由于对照组的背景表达，37个基因中的13个，包括HER-2、p53和BCL-2，在ctc中无法检测到表达。结论：从HRPC患者获得的CTCs及其基因表达谱可构建反义RNA文库。这可以增强HRPC的特征，并促进更有效治疗的发展。(C) 2004美国临床化学学会。
Background: Circulating tumor cells (CTCs) represent a surrogate source of tissue and conceptually represent a "real-time" biopsy. We previously reported that the number of CTCs mirrors disease progression in hormone-refractory prostate cancer (HRPC). To improve characterization of CTCs we further investigated whether in vitro transcription-based multigene reverse transcription-PCR expression profiles could be obtained from CTCs in HRPC.Methods: We evaluated the expression of 37 genes with potential utility for epithelial cell characterization from antisense RNA libraries constructed from immunomagnetically enriched CTCs from 7.5-mL blood samples from healthy donors and patients with HRPC.Results: In the control group 13 of 37 genes were not expressed. The most notable of the genes expressed in CTCs of 23 blood specimens drawn from 9 patients with metastatic prostate cancer were prostate-specific antigen (20 of 23; 87%), prostate-specific membrane antigen (17 of 23; 74%), androgen receptor (16 of 23; 70%), human glandular kallikrein 2 (7 of 23; 30%), epidermal growth factor receptor (4 of 23; 17%), and prostate-specific gene with homology to G protein receptor (2 of 23; 9%). The number of CTCs in these samples ranged from 4 to 283 in 7.5 mL of blood (mean, 87; median, 89). Expression of some of the genes was low in the control samples and higher in the patient samples. In all 23 samples, cytokeratin 19, epithelial cell adhesion molecule, or mucin 1 was expressed. Because of background expression in the controls, expression of 13 of the 37 genes, including HER-2, p53, and BCL-2, could not be measured in CTCs.Conclusion: Antisense RNA libraries can be constructed from CTCs and gene expression profiles of CTCs obtained from patients with HRPC. This could enhance the characterization of HRPC and facilitate the development of more effective therapies. (C) 2004 American Association for Clinical Chemistry.