A total of 49 HCV-infected patients who developed HCC (HCC group) were retrospectively examined. They were followed up (from 1988 to 2003) with an average period until HCC development being 6.5 ± 2.9 years. Paired serum samples at the time of chronic hepatitis C (pre-HCC sample) and HCC development (post-HCC sample) were collected. As a control group, 100 HCV-infected patients who were followed up over a period of 15 years (from 1988 to 2003) without
HCC development were retrospectively examined. Serum samples of the control group were available at the time of first visit to the clinic. All patients enrolled in this study were chronically infected with HCV genotype 1b (HCV-1b). HCV subtype was determined as reported previously. Serum HCV RNA titers BAY 57-1293 cost were quantitated by reverse-transcription polymerase chain reaction (RT-PCR0 with an internal RNA standard derived from the 5′ noncoding region of HCV (Amplicor HCV Monitor test, v. 2.0, Roche Diagnostics, Tokyo, Japan). All patients underwent liver
biopsy and were diagnosed as chronic hepatitis. All HCC and 68% (68/100) of non-HCC patients received IFN-monotherapy, either natural IFN alpha (Sumiferon, Dainipponsumitomo Pharmaceutical, Osaka, Japan) at a dose of 6 million units (MU) or recombinant HDAC inhibitors in clinical trials IFN alpha 2b (Intron A; Schering-Plough, Osaka, Japan) at a dose of 10 MU, 3 times a week for 6 months. All HCC patients were nonresponders (NR), who had detectable viremia during the entire
course of IFN treatment. On the other hand, 18 (26%) of the 68 non-HCC patients triclocarban treated with IFN achieved HCV RNA negativity at the end of treatment followed by rebound viremia within 6 months after the treatment and, therefore, they were referred to as relapsers. The other 50 IFN-treated, non-HCC patients were NR. The remaining 32 non-HCC patients did not receive IFN. All patients were seen every 2 months and tested for liver function markers during the follow-up period. HCV RNA was extracted from 140 μL of serum using a commercially available kit (QIAmp viral RNA kit; Qiagen, Tokyo, Japan). The core, NS3, and NS5A regions of the HCV genome were amplified as described elsewhere.[26, 32] The sequences of the amplified fragments were determined by direct sequencing. The aa sequences were deduced and aligned using GENETYX Win software version 7.0 (GENETYX, Tokyo, Japan). The numbering of aa was according to the polyprotein of the prototype of HCV-1b; HCV-J. Statistical differences in the baseline parameters of HCC and control groups were determined by Student’s t test for numerical variables and Fisher’s exact probability or chi-square tests for categorical variables. Likewise, statistical differences in viral mutations between HCC and control groups were determined by Fisher’s exact probability test.