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目的 研究外周血中胎蛋白信使核糖核酸(AFP mRNA)水平与原发性肝癌根治性切除术后复发转移的关系。方法 裸鼠肝内原位接种裸鼠人肝癌转移模型LCI-D20肿瘤组织,接种后第10天切除移植瘤,切除后第2天用不同剂量干扰素α-1b(IFNα-1b )治疗,治疗35 d后取外周血1 ml,用TaqMan实时定量逆转录－聚合酶链反应(RT-PCR)技术检测AFP mRNA水平,同时观察肿瘤复发转移情况。结果 对照组裸鼠术后肝内复发率和肺转移率均为100%(12/12),外周血AFP mRNA阳性率为100%。小剂量IFNα-1b治疗组肝内复发率62.5%(5/8),复发瘤体积小于对照组(25 mm~3±2mm~3对1143mm~3±3mm~3,t=9.27,P＜0.01),无肺转移,外周血AFP mRNA阳性率87.5%(7/8),水平低于对照组[(85±6)copies/μg对(955±2)copies/μg,t＝4.33,P＜0.01)。大剂量IFNα-1b治疗组肝内复发率为12.5%(1/8),体积仅0.5mm~,无肺转移,外周血AFP mRNA均阴性(x~2=11.67,P＜0.01)。结论 外周血AFP mRNA可作反映肝癌复发转移的敏感指标,TaqMan实时定量RT-PCR技术检测循环血肝癌细胞灵敏、简便、精确度高。
Objective To study the relationship between the level of fetoprotein messenger RNA (AFP mRNA) in peripheral blood and recurrence and metastasis after radical resection of primary liver cancer. Methods The nude mice were inoculated with LCI-D20 tumor tissue in situ into the liver of nude mice. The tumor was excised on the 10th day after inoculation and treated with interferon α-1b (IFNα-1b) on the second day after resection After 35 days, 1 ml of peripheral blood was taken and the level of AFP mRNA was detected by TaqMan real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). Meanwhile, the recurrence and metastasis of tumor was observed. Results The intrahepatic recurrence rate and lung metastasis rate in control group were 100% (12/12) and the positive rate of AFP mRNA in peripheral blood was 100%. The intrahepatic recurrence rate was 62.5% (5/8) in low-dose IFNα-1b treatment group, and the volume of tumor recurrence was smaller than that in control group (25 mm 3 ± 2 mm 3 vs 1143 mm 3 ± 3 mm 3, t 9.27, P 0.01 ), No metastasis, the positive rate of AFP mRNA in peripheral blood was 87.5% (7/8), which was lower than that of the control group [(85 ± 6) copies / μg vs (955 ± 2) copies / μg, t = 0.01). High-dose IFNα-1b treatment group, intrahepatic recurrence rate was 12.5% (1/8), the volume of only 0.5mm ~, no lung metastasis, peripheral blood AFP mRNA were negative (x ~ 2 = 11.67, P <0.01). Conclusion AFP mRNA in peripheral blood can be used as a sensitive index to reflect the recurrence and metastasis of hepatocellular carcinoma. TaqMan real-time quantitative RT-PCR detection of circulating hepatoma cells is sensitive, simple and accurate.