三对引物聚合酶链反应扩增检测肝细胞癌肝组织中乙型肝炎病毒脱氧核糖核酸

USING 3 PRIMER PAIRS TO DETECT HBV DNA IN LIVER TISSUES FROM HEPATOCELLULAR CARCINOMAS WITH PCR TECHNIQUE

用聚合酶链反应（ＰＣＲ）技术检测了１６例肝细胞癌（ＨＣＣ）患者癌灶和癌旁肝组织中乙型肝炎病毒脱氧核糖核酸（ＨＢＶ ＤＮＡ）。所用三对引物Ｓ１／Ｓ２、Ｃ１／Ｃ２和Ｘ１／Ｘ２分别选自ＨＢＶ ＤＮＡ的Ｓ区、前Ｃ和Ｃ区，以及Ｘ区。以质粒提取ＨＢＶＤＮＡ作灵敏度测定，ＰＣＲ－ＥＢ可达１０－２ｐｇ，ＰＣＲ－ＳＢＨ可达１０－６ｐｇ。以Ｓ、Ｃ和Ｘ引物扩增，阳性率分别为４３．８％（１４／３２） ７１．９％（２３／３２）和７１．９％（２３／３２）；以Ｃ引物和Ｘ引物扩增，阳性率差异无显著意义（Ｐ＞０．０５）。Ｓ引物扩增，阳性率与Ｃ引物比较，差异有显著意义（Ｐ＜０．０５）；与Ｘ引物比较，差异颇大（０．１０＞Ｐ＞０．０５）。１６例肝中均检出ＨＢＶＤＮＡ片段。结果提示，防治ＨＢＶ感染为我国ＨＣＣ一级预防的重要方面。Ｘ基因整合导致细胞恶变，Ｃ基因表达受阻逃避宿主的免疫监视可能是临床肝癌发生的机制。

Abstract PCR technique was used to detect HBV DNA in liver tissue samples for study of the preva-lence of HBV DNA in tumorous and nearby nontumorous liver tissues from 16 hepatocellularcarcinoma (HCC)patients.Three primer pairs, S1/S2, C1/C2 and X1/X2,used in this study were se-lected from S region, pre C and C region,and X region of HBV DNA, respectively. The detectionlimit with agarose gel electrophoresis and ethidium bromide staining (PCR-EB) was 10-2 pg, andwith Southern blot hybridization was 10-6 pg. The positive rates in amplification of HBV DNAby S, C and X regions primer pairs in liver samples were 43.8%(14/32),71.9% (23/32) and71.9%(23/32), respectively. There was significant difference between the the positive rates inamplification with S primer and with C primer (P<0.05),but no significant difference between the C primer and the X primer (P>0.05). and between the S primer and the X primer (0.10>P>0.05). HBV DNA fragments were detected in the livers from all 16 cases. The results indicatedthat x gene integration which induces hepatocellular carcinogenesis and arrest of C gene expression which evades host immune surveillance are the possible mechanisms of HCC development in pa-tients with persistent HBV infection.