肝癌病人手术期间不同部位血液AFP mRNA定量检测及其意义探讨

目的 研究肝细胞性肝癌(HCC)病人手术期间不同部位血液甲胎蛋白信使核糖核酸(AFP mRNA)的水平变化，分析其与临床病理学特征和复发转移的关系。方法 18例HCC和6例非HCC肝肿瘤病人，于切除肿瘤前后抽取门静脉、肝静脉和外周静脉血各5m1，肝炎、肝硬化病人各10例取外周血5m1。应用TaqMan实时定量逆转录—聚合酶链反应(RT—PCR)检测AFP mRNA水平。结果 术前外周血AFP mRNA相对HCC的灵敏度为72．2％，特异性为76.9％，准确率为75．0％。HCC病人术后各部位血液AFP mRNA水平明显高于术前相应部位水平。血液AFP mRNA水平与肿瘤大小、分化程度、有无肝内播散、包膜完整性以及血清AFP浓度均无关，而有门静脉癌栓的病人术后肝静脉血AFP mRNA水平明显高于无癌栓者。术后2个月内复发者，术后门静脉和外周静脉血AFP mRNA水平明显高于末复发者。结论 TaqMan实时定量RT—PCR法检测HCC病人血液AFP mRNA有较高的敏感性和特异性。手术可能促进部分HCC细胞和肝细胞脱落入血。动态监测外周血AFP mRNA水平对术后早期复发有预测价值。

Quantitative analysis of AFP mRNA in portal, hepatic and peripheral venous blood of patients with HCC during liver surgery and its clinical significance

Objective To study the changes of AFP mRNA expression in portal, hepatic and peripheral venous blood of patients with HCC during liver surgery and explore its relation to clinicopathological features and postoperative occurrence of HCC. Methods Portal, hepatic and peripheral venous blood samples were collected before and after tumor resection in 18 patients with HCC and 6 with non-HCC liver neoplasms. Meanwhile, peripheral blood samples were obtained from 20 patients with hepatitis/cirrhosis. AFP mRNA levels in all blood sample were measured using TaqMan real-time RT-PCR. Clinicopathological information of HCC patients was obtained during the follow-up period of 5-31 months. Results The positivity and levels of AFP mRNA in peripheral blood were significantly lower in patients with hepatitis/cirrhosis than in those with HCC. AFP mRNA was not detected in patients with non-HCC liver neoplasms before resection, while 50% positivity after resection, but the levels were lower than in patients with HCC. In those patients with HCC, the positivity of AFP mRNA in portal, hepatic and peripheral venous blood before resection was 72.2%, 77.8% and 72.2%, respectively, but all 100% after resection. Meanwhile, the levels markedly increased after resection. The levels of AFP mRNA in all blood samples were not correlated with tumor size, histological grade, intrahepatic dissemination, capsule integrity or serum AFP concentration. The levels in tumor-draining hepatic venous blood after resection were higher in HCC patients with portal venous thrombosis than in those without thrombosis. Furthermore, the levels of AFP mRNA in portal and peripheral venous blood after resection were higher in HCC patients who had intrahepatic recurrence in 2 months postoperatively. Conclusions Quantitative analysis of AFP mRNA by TaqMan real-time RT-PCR is a very sensitive and specific method for detecting circulating HCC cells. HCC cells and some hepatocytes are released preoperatively and intraoperatively into the blood circulation, which may potentially be the source of recurrence or metastasis. Sequential quantification of AFP mRNA in peripheral blood may predict impending recurrence, especially in 2 months after operation.