巢式RT-PCR检测乳腺癌外周血循环肿瘤细胞的临床价值
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江苏省六大人才高峰项目(06-B-070)


Clinical value of detection of circulating tumor cells of breast cancer by nested RT-PCR
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    摘要:

    目的:联合采用细胞角蛋白19(cytokeratin 19,CK19)及乳腺上皮黏蛋白(small breast epithelial mucin,SBEM)指标,探讨通过巢式RT-PCR(nested RT-PCR)方法检测乳腺癌患者外周血循环肿瘤细胞(circulating tumor cells,CTC)的临床价值-方法:采用MCF-7 RNA与健康志愿者外周血单核细胞(peripheral mononuclear cell,PMNC)RNA进行梯度混合后的反转录产物,检测优化后巢式RT-PCR体系的敏感性,再选择10例健康志愿者以及20例乳腺良性疾病患者外周血检测其特异性后,对56例乳腺癌患者外周血中CK19 mRNA及SBEM mRNA表达情况进行检测- 结果:优化后的检测体系能够在3 ml外周血PMNCs中检测到1个MCF-7细胞,10例健康志愿者以及20例乳腺良性疾病患者外周血中未检测到CK19 mRNA与SBEM mRNA的表达;两者的表达与原发肿瘤大小,腋窝淋巴结状况及TNM分期相关(P < 0.05),而与原发肿瘤病理类型,激素受体以及C-erbB-2的表达无明显相关性(P > 0.05)- 结论:经过优化的巢式RT-PCR体系联合检测乳腺癌患者外周血CK19 mRNA及SBEM mRNA的表达敏感性和特异性较高,能用于检测外周血CTC,对进一步精确评估预后及指导治疗可能有较高的临床价值-

    Abstract:

    Objective:To evaluate the clinical value of detection of circulating tumor cells of breast cancer by nested RT-PCR using combimation of cytokeratin 19(CK19)and small breast epithelial mucin(SBEM). Methods:The reverse transcripted products of the gradient mixture of the RNA of MCF-7 cells and peripheral mononuclear cells of healthy volunteers were detected by the optimized nested RT-PCR to evaluate its sensitivity. Peripheral blood of 10 healthy volunteers and 20 patients of benign lesions of breast were detected to evaluate its specificity,and peripheral blood of 56 patients of breast cancer were detected. Results:The optimized detecting system detected MCF-7 cell from peripheral mononuclear cells of 3 ml peripheral blood,and no positive expression of CK19 mRNA and SBEM mRNA were found in 10 healthy volunteers and 20 patients of benign lesions of breast. The expression of the two markers was related to the size of primary tumor,axillary lymph nodes status and TNM(P < 0.05),but not related to the status of ER,PR and C-erbB-2(P > 0.05). Conclusion:The optimized nested RT-PCR system achieved higher sensitivity and specificity,could detect the expression level of CK19 mRNA and SBEM mRNA effectively in the peripheral blood of breast cancer patients;CTC detection is useful for the assessment of prognosis and treatment of breast cancer patients.

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傅〓卫,王〓水,杜〓青,肇〓毅,查小明,刘晓安.巢式RT-PCR检测乳腺癌外周血循环肿瘤细胞的临床价值[J].南京医科大学学报(自然科学版),2010,(6):818-821858

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  • 收稿日期:2010-02-09
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