SYBR Green I实时荧光定量方法检测结直肠癌患者外周血CEA-mRNA表达研究  

Study of CEA mRNA expression in colorectal cancer patients peripheral blood with SYBR Green I real-time quantitative polymerase chain reaction

作　　者：闫先侠 王传新[2] 牛爱军[3] 王燕[3] 陈英剑[3] 

机构地区：[1]山东省精神卫生中心检验科,山东济南250014 [2]山东大学齐鲁医院检验科 [3]济南军区总医院实验诊断科

出　　处：《中国实验诊断学》2007年第9期1228-1231,共4页Chinese Journal of Laboratory Diagnosis

摘　　要：目的建立检测CEA mRNA表达水平的实时荧光定量逆转录聚合酶链反应(FQ-PCR)方法,探讨外周血CEA mRNA表达在结直肠癌患者中的应用价值。方法应用SYBR Green I作为荧光染料,以GAPDH作为内对照,建立检测CEA mRNA的FQ-PCR方法,并对56例结直肠癌患者外周血中CEA mRNA的表达水平进行了检测。结果56例结直肠癌患者中有33例外周血CEA mRNA表达为阳性,表达水平为1.07±2.99,阳性表达率为66.1%(37/56),30例结直肠良性疾病患者中有2例外周血CEA mRNA表达为阳性,表达水平为0.13±0.07,阳性表达率为6.7%(2/30),30例健康成人外周血CEA mRNA表达均为阴性;结直肠癌患者外周血CEA mRNA在不同病理分化程度中表达水平无明显变化(P>0.05),但在Dukes C+D期表达水平(1.73±2.15)及表达阳性率82.8%(24/29)均显著高于Dukes A+B期表达水平(0.39±0.87)及表达阳性率48.1%(13/27)(P<0.01)。结论该FQ-PCR方法简便、特异、可靠;检测外周血CEA mRNA表达水平,对诊断结直肠肿瘤病变比血清CEA蛋白具有更高的敏感性,可提高早期结直肠癌诊断符合率;在发生转移的Dukes C+D期其表达水平及表达阳性率均显著高于未发生转移的Dukes A+B期,可作为鉴别结直肠癌患者发生微转移的有力证据之一,动态观察其水平变化对判断结直肠癌患者的预后有重要价值。Objective To develop a real-time quantitative RT-PCR method for detection peripheral blood CEA mRNA expression in colorectal cancer patients.Methods Using SYBR Green I as dye and GAPDH as inner reference,we established FQ-PCR method and detected the CEA mRNA expression of peripheral blood in 56 colorectal cancer patients. Results 33 of all 56 colorectal cancer patients＇expressed CEA mRNA in peripheral blood, the expressed levels were 1.07 ± 2.99 and the and the positive ratio was 66.1% （37/56） ,2 of 30 colon benign patient expressed CEA mRNA in peripheral blood,tire expressed levels were 0.13 ± 0.07 and the positive ratio was 6.7% （2/30）, none of 30 normal control expressed CEA mRNA in peripheral blood. CEA mRNA levels in peripheral blood had no significant difference in pathology differentitation（ P 〉 0.05） ,but CEA mRNA levels （1.73 ± 2.15） and the positive ratio 82,8% （24/29） of Dukes C ＋ D stages were respectively significantly higher than that of Dukes A ＋ B stages which were （0.39 ± 0.87） and 48.1% （13/27） （P 〈 0.01 ）. Conclusion The electropharesis and melting curve analysis of gene amplification product showed that the FQ-PCR method is simple, specige and reliable. D CEA mRNA levels in peripheral blood had higher sensitivity, than CEA mRNA protein in diagnosing colorectal cancer and can improve correct diagnoses ratio. CEA mRNA and positive ratio in peripheral blood of Dukes C ＋ D stages were significantly higher than that of Dukes A ＋ B stages, CEA mRNA can be considered one of conclusive proof in differentiating mierometastastases and had important value in judging colorectal cancer prognosis.

关 键 词：结直肠癌 CEA MRNA 实时定量聚合酶链反应 

分 类 号：R735.3[医药卫生—肿瘤]