SYBR GREEN荧光定量聚合酶链反应检测粘蛋白1在乳腺癌诊断中的应用

The application of SYBR GREEN fluorescent quantification polymerase chain reaction for the detection of human mammaglobin mRNA in the diagnosis of breast cancer

目的探讨SYBR GREEN荧光定量聚合酶链反应法检测外周血循环肿瘤细胞中粘蛋白1基因表达水平在乳腺癌诊断和治疗监测中的应用。方法以β-actin为内对照,采用SYBR GREEN荧光定量聚合酶链反应法测定45名健康女性体检者、91例良性乳腺疾病患者和193例乳腺癌患者以及87例其它肿瘤患者外周血中粘蛋白1的表达量,分析不同临床病理因素与基因表达的相关性。结果乳腺癌患者粘蛋白1阳性率显著高于健康体检者、良性乳腺疾病和其它肿瘤患者(P<0.05);良性乳腺疾病及其它肿瘤患者粘蛋白1阳性率显著高于健康体检者(P<0.05),而良性乳腺疾病与其它肿瘤患者粘蛋白1阳性率差异无统计学意义(P>0.05);乳腺癌临床分期、组织学分级、腋淋巴结转移及HER2/neu表达是MUC1 mRNA表达独立相关因素(P<0.05)。结论 SYBR GREEN荧光定量聚合酶链反应法是灵敏较高、特异较强的快速定量检测粘蛋白1方法,可有效监测乳腺癌诊断、疗效、转移和预后。

Objective To investigate the application of SYBR GREEN fluorescent quantification polymerase chain reaction（ SYBR GREEN FQ-PCR） to detect the MUC1 mRNA of circulating tumor cells in diagnosis and therapeutic monitoring of breast cancer.Methods With β-actin as internal control,MUC1 mRNA in peripheral blood cell samples in 45 health women,91 patients with benign breast disease and 193 patients with breast cancer and 87 with other cancer patients were detected by SYBR GREEN FQ-PCR.The relationship between MUC1 mRNA and clinicopathological parameters was analyzed.Results The positive rate of MUC1 mRNA detection in the patients with breast cancer was significantly higher than that in healthy individuals,patients with benign breast disease patients and other types of cancers（ P 〈0. 05）.The positive rate in the patients with benign breast disease and other types of cancers was significantly higher than that in the healthy individuals（ P〈 0. 05）.There was no significant difference in the positive rate between benign breast disease patients and other cancer patients（ P〉0. 05）. The independent factors of MUC1 mRNA expression were clinical stage,histological grade and axillary metastatic lymph node and HER2 / neu（ P〈 0. 05）.Conclusion FQ-PCR is a rapid and sensitive and specific method for quantitative detection of mucin1 mRNA. It can be used for the diagnosis,therapy,metastasis and prognosis of breast cancer.