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目的:筛选一组可检测乳腺癌循环癌细胞的标记物基因,分析乳腺癌患者相对循环癌细胞数(Lc)与临床病理特征的关联。方法:收集乳腺癌患者142例(乳腺癌组)和正常女性60人(正常对照组)外周血标本。利用CGAP提供的SAGE Genie数据库中数字基因表达演示工具,筛选一组可用于检测乳腺癌循环癌细胞的标记物基因(FAM83A、NPY1R和KRT19)。应用定量巢式PCR方法检测研究对象外周血中标记物基因的表达水平,并通过公式计算患者Lc值。结果:在乳腺癌组患者外周血中肿瘤标记物基因FAM83A、NPY1R和KRT19的表达水平均明显高于正常对照组(P<0.001)。142例乳腺癌患者中有113例(79.6%)至少表达1个肿瘤标记物基因。乳腺癌患者Lc值与其临床分期和远处转移有关联(P<0.001)。Kaplam-Meier生存曲线,Lc值小于2的患者生存时间明显长于Lc值大于2者(P=0.005)。结论:筛选了一组检测乳腺癌外周血循环癌细胞的标记物基因,联合该组基因计算的Lc值与患者临床分期和远处转移有关联,并可辅助判断乳腺癌预后。
OBJECTIVE: To screen a panel of markers that can detect circulating cancer cells in breast cancer and to analyze the relationship between the relative circulating tumor cells (Lc) and clinicopathological features in patients with breast cancer. Methods: Peripheral blood samples were collected from 142 breast cancer patients and 60 normal women (normal control group). A set of marker genes (FAM83A, NPY1R and KRT19) that can be used to detect circulating breast cancer cells was screened using the digital gene expression demonstration tool provided by CGAP in the SAGE Genie database. Quantitative nested PCR was used to detect the expression level of marker genes in the peripheral blood of the study subjects and the patients’ Lc values were calculated by the formula. Results: The expression of tumor marker gene FAM83A, NPY1R and KRT19 in peripheral blood of patients with breast cancer was significantly higher than that of the normal control (P <0.001). 113 of 142 breast cancer patients (79.6%) expressed at least one tumor marker gene. Lc values in breast cancer patients were correlated with their clinical stage and distant metastasis (P <0.001). Kaplam-Meier survival curves, patients with Lc values less than 2 were significantly longer than those with Lc values greater than 2 (P = 0.005). CONCLUSIONS: A panel of marker genes for detecting circulating breast cancer cells in peripheral blood was screened. The Lc values calculated with this group of genes were correlated with clinical stage and distant metastasis, and could be helpful in predicting the prognosis of breast cancer.