荧光定量PCR检测外周血中JAR细胞的探讨

Study of fluorescence quantitative polymerase chain reaction in detection of JAR cells from peripheral blood

目的 建立灵敏可靠的外周血中检出滋养细胞肿瘤细胞的方法，为早期诊断滋养细胞肿瘤的转移奠定实验基础。 方法 在已知数量的ＪＡＲ细胞掺入１０ｍｌ外周血中建立肿瘤细胞血行转移实验模型，荧光定量逆转录－聚合酶链反应（ＦＱ－ＲＴ－ＰＣＲ）定量检测ＪＡＲ细胞的 β－ｈＣＧ—ｍＲＮＡ。结果 ＦＱ-ＲＴ－ＰＣＲ可检测到相当于１个ＪＡＲ细胞表达的 β－ｈＣＧ—ｍＲＮＡ量，但当ＪＡＲ细胞被掺入１０ｍｌ外周血中时，细胞数＞１０２个方可经富集后可检测到。结论 ＦＱ－ＲＴ－ＰＣＲ是一种敏感的检测手 段，有希望成为检测外周血中滋养细胞肿瘤细胞的敏感方法，将有助于滋养细胞肿瘤转移病例的早期诊断与治疗。

Objective To found sensitive and reliable method to detect trophoblastic tumor cells from peripheral blood, establish experimental method for early diagnosis of metastasis of gestation trophoblastic tumor (GTT). Methods Known numbers of JAR cells were mixed into 10ml Peripheral blood as a model for trophoblastic tumor cells in the peripheral blood. Quantitative detection of β-hcG mRNA expression of JAR cells by fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCCR) were performed. Results FQ-RT-PCR could detect β-hCG mRNA equal to one JAR cell expression. It could be detected>102 trophoblasts when mixed into maternal blood after enrichment. Conclusions The method used in this research proved very sensitive in detection of hCG-mRNA expression cells, and it would be used in detecting trophoblastic tumor cells and benefit finding early metastasis of GTT.