巢式RT-PCR技术检测肺癌76例微量癌细胞转移分析

Detection of micrometastases of lung cancer in peripheralblood and lymph node by nested RT-PCR

目的 :建立检测肺癌患者外周血及淋巴结微转移癌细胞的敏感的分子检测方法 ,并探讨其临床应用的意义。方法 :采用巢式逆转录聚合酶链反应 ( RT- PCR)技术特异引物扩增 CK19- m RNA,以检测肺癌患者外周血及淋巴结中 CK19- m RNA的表达情况。分析CK19- m RNA扩增结果与患者临床指标的关系。结果 :76例肺癌患者外周血标本中 39例CK19- m RNA阳性 ,阳性率为 5 1 .3% ,2 7例肺良性病变中仅 1例阳性 ,阳性率为 3.7% ,1 5例健康对照者均为阴性。全部患者 1 5 7枚淋巴结中 CK19- m RNA检测出阳性率为 38.9%( 61 / 1 5 7) ,常规病理方法检出阳性率为 1 4.6% ( 2 3/ 1 5 7) ,两种方法比较 ,有显著性差异 ( P<0 .0 5 ) ,且 HE染色阴性的 1 34枚淋巴结中 ,经 RT- PCR法证实存在癌转移的阳性率为 2 8.3% ( 38/ 1 34) ,两组比较有显著性差异 ( P<0 .0 5 )。结论 :巢式 RT- PCR是一种特异性和敏感性均较高的癌细胞微量转移检测方法 ;可能有助于肺癌细胞微转移的早期诊断。

Objective: To establish a sensitive method of detecting micrometastasis in peripheral blood and lymph nodes of lung cancer and study its clinical implication.Methods: Nest RT-PCR was used to detect CK 19-mRNA expression in peripheral blood and lymph node of lung cancer.Results:Of 76 lung cancer patients,39(51.3%)had positive CK 19-mRNA, while CK 19-mRNA was detected only 1 in 27 patients with pulmonary benign lesion and was not detected among 15 healthy volunteers.The positive rate of the CK 19-mRNA express in lymph nodes of all 157 patients is 38.9%(61/157) by nest RT-PCR,while the positive rate is 14.6%(23/157)by traditional pathological method(P<0.05).Of 134 lymph nodes which were detected negative by HE dye,38 cases were detected positive by nest RT-PCR.The difference is obvious.Conclusion :Nest RT-PCR technique is a specific and sensitive method to detect micrometastasis in patients with lung cancer.It may be very useful in diagnosing micrometastasis earlier.