Detection of Androgen Receptor Mutations in Circulating Tumor Cells in Castration-Resistant Prostate Cancer
Detection of Androgen Receptor Mutations in Circulating Tumor Cells in Castration-Resistant Prostate Cancer
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DOI：
10.1373/clinchem.2010.143297
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发表时间：
2010-09-01
影响因子：
9.3
通讯作者：
Gross, Mitchell E.
中科院分区：
文献类型：
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作者：
Jiang, Yuqiu;Palma, John F.;Gross, Mitchell E.
背景技术背景：已在晚期前列腺癌患者的组织样本中鉴定出AR（雄激素受体）基因的编码突变，并代表了去势抵抗性前列腺癌（CRPC）发生的可能机制。可用于CRPC患者分子研究的肿瘤来源组织很少。CRPC患者血液中的循环肿瘤细胞（CTC）代表了询问这类患者疾病的可能途径。方法：用CellSearch（R）循环肿瘤细胞（CTC）试剂盒和CellSearch Profile试剂盒加Qiagen的AllPrep DNA/RNA Micro试剂盒捕获循环肿瘤细胞，分别用于测量每7.5mL血液的CTC计数和用于分离核酸。结果：35例CRPC患者中20例检测到AR基因突变，其中错义突变19例，沉默突变2例，缺失突变5例，插入突变1例。扩增产物中突变体的相对丰度在5%~ 50%之间。许多AR突变在手术活检或尸检中被确定，并与雄激素导向therapy.CONCLUSIONS：AR突变可以在CTC富集的外周血样本CRPC患者。这种方法有可能为理解CTCs以及CRPC中肿瘤进展和转移的机制开辟新的视角。
BACKGROUND: Coding mutations in the AR (androgen receptor) gene have been identified in tissue samples from patients with advanced prostate cancer and represent a possible mechanism underlying the development of castration-resistant prostate cancer (CRPC). There is a paucity of tumor-derived tissue available for molecular studies of CRPC patients. Circulating tumor cells (CTCs) in the blood of CRPC patients represent a possible avenue for interrogating the disease of such patients.METHODS: Circulating tumor cells were captured with the CellSearch (R) Circulating Tumor Cell (CTC) Kit and with the CellSearch Profile Kit plus Qiagen's AllPrep DNA/RNA Micro Kit for the measurement of the CTC count per 7.5 mL of blood and for the isolation of nucleic acids, respectively. The AR gene was amplified by the PCR, and mutation status and relative abundance were analyzed by applying Transgenomic's WAVE (R) denaturing HPLC technology followed by direct sequencing.RESULTS: AR mutations were detected in 20 of 35 CRPC patients; 19 missense mutations, 2 silent mutations, 5 deletions, and 1 insertion were observed. The relative abundance of the mutants in the amplified products ranged from 5% to 50%. Many of the AR mutations were identified in surgical biopsies or at autopsy and were associated with resistance to androgen-directed therapies.CONCLUSIONS: AR mutations can be identified in CTC-enriched peripheral blood samples from CRPC patients. This approach has the potential to open new perspectives in understanding CTCs and the mechanisms for tumor progression and metastasis in CRPC.