巢式逆转录多聚酶链反应检测循环性肝癌细胞及其临床意义

The detection of circulating hepatocellular carcinoma cells in peripheral venous blood by reverse transcription polymerase chain reaction and its clinical significance

目的应用巢式逆转录多聚酶链反应（ＲＴＰＣＲ）检测血液中播散的循环性肝癌细胞并探讨其意义。方法抽取９３例肝癌患者及３７例对照组患者（１５例肝炎后肝硬变患者，１２例慢性肝炎患者，１０例健康人）周围静脉血液各５ｍｌ，应用巢式ＲＴＰＣＲ检测其周围静脉血液中ＡＦＰｍＲＮＡ。结果共有５０个肝癌血液标本被检测到ＡＦＰｍＲＮＡ（５３．８％），而对照组中无一例血液中检测到ＡＦＰｍＲＮＡ，ＡＦＰｍＲＮＡ检出率与肝癌分期（ＴＮＭ）、血清ＡＦＰ浓度、肝内转移、门静脉癌栓、肿瘤大小及远处转移有明显联系（Ｐ＜００５）。２１例伴有肝外远处转移的原发性肝癌血液标本均被检测到ＡＦＰｍＲＮＡ（１００％），没有远处转移的７２例肝癌血液标本中２９例检测到ＡＦＰｍＲＮＡ（４０３％）。结论ＡＦＰｍＲＮＡ可以作为肝癌细胞血液转移的标志，也可作为肝癌远处转移的预测性指标之一。

Objective To detect circulating hepatocellular carcinoma by demonstrating hepatocellular carcinoma cells associated mRNA in the nuclear cell component of peripheral blood (PBL). Method Peripheral blood (5ml) samples were obtained from 93 patients with hepatocellular carcinoma (HCC) and from 37 controls (15 contrds with liver cirrhosis after hepatitis B, 12 chronic hepatitis B,and 10 normal liver function). To identify HCC cells in peripheral blood, liver specific human alpha fetoprotein (AFP) mRNA was amplified from total RNA extracted from whole blood by reverse transcription polymerase chain reaction. Result AFP mRNA was detected in 50 blood samples from the patients with HCC (53 8%). In contrast,there were no clinical control patientswhose samples showed detectable AFP mRNA in PBL. The presence of AFP mRNA in blood seemed to be correla ted with the stage (by TNM classification) of HCC, the serum AFP value, and the presence of intrahepatic metastasis, portal vein thrombosis, tumor diameter and /or distant metastasis.AFP mRNA was detected in the blood of 21 patients showing metastasis at extrahepatic organs (100%) in contrast to 29 of 72 patients without metastasis (40 3%). Conclusion The presence of AFP mRNA in peripheral blood may be an indicator of malignant he patocytes, which might predict hematogenous spreading metastasis of tumor cells in patients with HCC.