Assessment of a six gene panel for the molecular detection of circulating tumor cells in the blood of female cancer patients.
Assessment of a six gene panel for the molecular detection of circulating tumor cells in the blood of female cancer patients.
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DOI：
10.1186/1471-2407-10-666
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发表时间：
2010-12-03
期刊：
影响因子：
3.8
通讯作者：
Zeillinger R
中科院分区：
文献类型：
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作者：
Obermayr E;Sanchez-Cabo F;Tea MK;Singer CF;Krainer M;Fischer MB;Sehouli J;Reinthaller A;Horvat R;Heinze G;Tong D;Zeillinger R
癌症患者外周血中循环肿瘤细胞(CTC)的存在已经被描述为各种实体肿瘤，并且它们的临床相关性已经被证明。原发肿瘤的遗传异质性和上皮性抗原的缺失可能会阻碍基于上皮性抗原分析的CTC检测。因此，我们的目标是寻找新的基因标记物，用于检测女性癌症患者的CTC。应用基因芯片技术检测了38个癌细胞株(乳腺、卵巢、宫颈和子宫内膜)和10例健康女性捐献者外周血单个核细胞(PBMC)的基因表达。用最大T检验和50%单侧剪裁最大T检验鉴定差异表达基因。使用AB TaqMan®低密度阵列对380个基因靶点进行验证性RT-qPCR。然后，使用相同的RT-qPCR平台对126例乳腺癌、卵巢癌和子宫内膜癌患者的肿瘤组织中的93个基因靶点进行分析。最后，26名健康女性和125名患者(原发性乳腺癌、卵巢癌、宫颈癌或子宫内膜癌，以及晚期乳腺癌)的血液样本在OncoQuick浓缩和RNA预扩增后进行了分析。同样，分析了乳腺癌和卵巢癌患者血液中hMAM和EpCAM基因的表达。对于每个基因，将截断阈值设置为与健康对照组的平均表达水平的三个标准偏差，以确定潜在的CTC检测标记。在81%的晚期乳腺癌患者和29%的原发性乳腺癌患者的血液样本中，有6个基因过表达。EPCAM基因表达分别为19%和5%，而HMAM基因表达仅见于晚期组(39%)。使用新的六基因组合的多标记分析对44%的宫颈、的子宫内膜和19%的卵巢癌患者进行了阳性诊断。在检测乳腺癌血液中循环肿瘤细胞方面，6个基因的组合被发现优于EpCAM和HMAM，它们可能成为子宫内膜癌、子宫颈癌和卵巢癌CTC的潜在标记物。
The presence of circulating tumor cells (CTC) in the peripheral blood of cancer patients has been described for various solid tumors and their clinical relevance has been shown. CTC detection based on the analysis of epithelial antigens might be hampered by the genetic heterogeneity of the primary tumor and loss of epithelial antigens. Therefore, we aimed to identify new gene markers for the PCR-based detection of CTC in female cancer patients. Gene expression of 38 cancer cell lines (breast, ovarian, cervical and endometrial) and of 10 peripheral blood mononuclear cell (PBMC) samples from healthy female donors was measured using microarray technology (Applied Biosystems). Differentially expressed genes were identified using the maxT test and the 50% one-sided trimmed maxT-test. Confirmatory RT-qPCR was performed for 380 gene targets using the AB TaqMan® Low Density Arrays. Then, 93 gene targets were analyzed using the same RT-qPCR platform in tumor tissues of 126 patients with primary breast, ovarian or endometrial cancer. Finally, blood samples from 26 healthy women and from 125 patients (primary breast, ovarian, cervical, or endometrial cancer, and advanced breast cancer) were analyzed following OncoQuick enrichment and RNA pre-amplification. Likewise, hMAM and EpCAM gene expression was analyzed in the blood of breast and ovarian cancer patients. For each gene, a cut-off threshold value was set at three standard deviations from the mean expression level of the healthy controls to identify potential markers for CTC detection. Six genes were over-expressed in blood samples from 81% of patients with advanced and 29% of patients with primary breast cancer. EpCAM gene expression was detected in 19% and 5% of patients, respectively, whereas hMAM gene expression was observed in the advanced group (39%) only. Multimarker analysis using the new six gene panel positively identified 44% of the cervical, 64% of the endometrial and 19% of the ovarian cancer patients. The panel of six genes was found superior to EpCAM and hMAM for the detection of circulating tumor cells in the blood of breast cancer, and they may serve as potential markers for CTC derived from endometrial, cervical, and ovarian cancers.
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影响因子：
45.3
作者：
Cristofanilli, M;Hayes, DF;Terstappen, LWMM
通讯作者：
Terstappen, LWMM
影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
3.8
作者：
Mikhitarian K;Martin RH;Ruppel MB;Gillanders WE;Hoda R;Schutte del H;Callahan K;Mitas M;Cole DJ
通讯作者：
Cole DJ
影响因子：
12.3
作者：
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者：
Zhang J
影响因子：
8.8
作者：
通讯作者：
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