妊娠滋养细胞肿瘤患者外周血肿瘤细胞检测方法的研究

Detection of tumor cells in the peripheral blood of the patients suffered from gestational trophoblastic tumor

目的研究从妊娠滋养细胞肿瘤患者外周血中检出肿瘤细胞的方法。方法具有βhCGmRNA表达特征的绒癌细胞株(JAR细胞)掺入正常人外周血中,用荧光定量逆转录聚合酶链反应检测其中βhCGmRNA,推测JAR细胞的量;采集治疗前的妊娠滋养细胞肿瘤患者外周血,定量检测其中的βhCGmRNA,以推测滋养细胞的含量。结果当10ml外周血中含有102以上JAR细胞时可检测到其中的βhCGmRNA;9例滋养细胞肿瘤中有4例的外周血中检测到βhCGmRNA表达。推算肿瘤细胞量为104～107个/10ml外周血。结论荧光定量RTPCR法测定βhCGmRNA是检测、推算外周血中滋养细胞肿瘤细胞的有用方法之一;部分妊娠滋养细胞肿瘤患者外周血中可检测到肿瘤细胞。

Objective To found sensitive and reliable method to identify trophoblastic tumor cells in the peripheral blood of the patients suffered from gestational trophoblastic tumor.Methods Given numbers of JAR cell from ten to million were mixed into 10ml non pregnant peripheral blood as a model. Detection of β hCG mRNA with fluorescence quantitative reverse transcription polymerase chain reaction (FQ RT PCR) and then estimation of the numbers of tumor cell in the blood. Nine cases of peripheral blood were collected from the pretreatment patients of gestational trophoblastic tumor to assay β hCG mRNA with FQ RT PCR, then to estimate the numbers of tumor cell in the circulation blood. Results FQ RT PCR could detect β hCG mRNA when ≥10 2 JAR cells were mixed into 10ml non pregnant peripheral blood. Four cases of bloods had been detected β hCG mRNA expression in 9 cases of gestational trophoblastic tumor, and the numbers of tumor cell from 10 4 to 10 7 per 10ml blood. Conclusion FQ RT PCR of which primers and probe are designed for β hCG had been proved to be very sensitive detection means, it could be used to detect gestational trophoblastic tumor cells from patient preipheral blood; With FQ RT PCR the tumor cells had been detected in some patients of gestational trophoblastic tumor.