title: HBx-DNA probe preparation and its application in study of hepatocarcinogenesis.
abstract: AIM:To study the role of HBV especially HBx Open Reading Frame (ORF) in the development of hepatocellular carcinoma (HCC).METHODS:HBV 3.2kb fragment was retrieved by digesting recombinant plasmid pBR(322)-2HBV with EcoR II, and HBx 0.59kb fragments by digesting HBV-DNA with BamH I and Bgl II. These fragments were labelled with digoxigenin to get HBV-DNA and HBx-DNA probes. HBV-DNA was detected in HCC by dot blot and Southern blot hybridization with HBV-DNA probe, so the positive specimens in which HBV-DNA were integrated were selected. HBx-DNA was subsequently detected in the selected specimens with HBx-DNA probe.RESULTS:HBV-DNA was detected in 75% HCC, among which integrated type, integrated + free type covered 63.6% and 36.4%. There was no free type. HBx-DNA was detected in 90.5% specimens of integrated type.CONCLUSION:Hepatocarcinogenesis was highly related to HBV-DNA integration, and HBV-DNA mainly integrated into chromosome with incomplete virus DNA fragments among which HBx fragment was the predominant one.
authors.forename: Feng-Guang
authors.initials: FG
authors.lastname: Gao
authors.forename: Wen-Sheng
authors.initials: WS
authors.lastname: Sun
authors.forename: Ying-Lin
authors.initials: YL
authors.lastname: Cao
authors.forename: Li-Ning
authors.initials: LN
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Song
authors.forename: Hua-Fen
authors.initials: HF
authors.lastname: Li
authors.forename: Shi-Kun
authors.initials: SK
authors.lastname: Yan