【摘要】 目的　应用ＲＴ－ＰＣＲ技术研究原发性肝癌患者外周血单个核细胞（ＰＢＭＣ）甲胎蛋白（ＡＦＰ）ｍ ＲＮＡ 表达与肝癌转移的关系。方法　原发性肝癌４８ 例。慢性肝病患者２１ 例。正常对照２０ 例。采外周血，肝素抗凝，分离单个核细胞及分离总ＲＮＡ。逆转录酶合成ｃＤＮＡ。设计扩增ＡＦＰ基因的引物，进行巢式ＰＣＲ扩增。结果　正常人ＰＢＭＣ均无ＡＦＰｍ ＲＮＡ 表达。原发性肝癌患者的ＰＢＭＣＡＦＰｍ ＲＮＡ阳性率（２８／４８，５８．３３％ ）明显高于慢性肝病组（３／２１，１４．２８％ ，Ｐ＜ ０．０１）。伴远处转移的原发性肝癌患者全部为阳性（８／８）。结论　ＲＴ－ＰＣＲ检测原发性肝癌患者ＡＦＰｍ Ｒ－ＮＡ 有可能成为反映肝癌细胞血行播散的指标。更多还原

【Abstract】 Objective A nested reverse transcription polymerase chain reaction(RT PCR) was developed to study the relationship between expression of AFP mRNA in peripheral blood mononuclear cells and metastasis in patients with hepatocellular carcinoma(HCC).Methods Heparinized blood samples were obtained from 48 patients with HCC,21 patients with chronic liver diseases and 20 normal subjects.Mononuclear cells were separated from blood.Total RNA was extracted by the acid guanidium phenol chloroform method.The first strand of cDNA was synthesized from an aliquot of the RNA using Moloney murine leukemia virus reverse transcriptase and random hexamers.The two pairs of primers chosen from the sequences of AFP were used for the two step PCR.Results AFP mRNA in peripheral blood cells was not detected in all normal subjects, but it was detecte in 28 of 48 (58.33%) cases with HCC and in all 8 patients with extrahepatic metastasis.The positive rate in patients with HCC was significantly higher than the cases with chronic liver diseases(3/21,14.28%, P <0.01).Conclusions The detection of AFP mRNA in peripheral blood cells may be helpful for clinial detection of metastasis of HCC.$$$$更多还原