Detection and analysis of replicating hepatitis C virus RNA in hepatocellular carcinoma tissues.
Although persistent hepatitis C virus infection is closely associated with the development of hepatocellular carcinoma, the nature of hepatitis C virus replication in the hepatocellular carcinoma tissue has not been fully characterized. To study this, carcinoma and non-carcinoma tissues were obtained from five patients with hepatocellular carcinoma. Total RNA was recovered from each tissue, and a portion of the envelope gene of replicating hepatitis C virus was amplified by minus-strand-specific reverse transcription and nested polymerase chain reaction. The amplified cDNA was examined by single strand conformation polymorphism analysis and sequencing. Hepatitis C virus replication was detected in both carcinoma and non-carcinoma tissues in four patients who were positive for serum hepatitis C virus markers. In one patient, a single species with identical envelope 2 genome was obtained from both carcinoma and non-carcinoma tissues. In the other three patients, the replicating hepatitis C virus existed as a mixture of 2-5 species with different but highly homologous (82-99%) envelope 2 genomes (quasispecies populations). The constitution of viral populations was different between carcinoma and non-carcinoma tissues. A total of ten sequences were recovered; four sequences were found in both tissues, two were found in carcinoma tissues, and four were found in non-carcinoma tissues. The difference in the constitution of quasispecies populations between carcinoma and non-carcinoma tissues confirms the unequivocal replication of hepatitis C virus in both tissues, and may imply the presence of different biological properties among hepatitis C virus with different sequences.
Kurosaki
M
M
Second Department of Internal Medicine, Faculty of Medicine, Tokyo Medical and Dental University, Japan.
Enomoto
N
N
Sakamoto
N
N
Tanaka
Y
Y
Tang
L
L
Hoshino
Y
Y
Izumi
N
N
Marumo
F
F
Sato
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000821:virology*; D016174:Hepacivirus / Q000302:isolation & purification*; D006801:Humans; D008113:Liver Neoplasms / Q000821:virology*; D008969:Molecular Sequence Data; D018807:Polymorphism, Single-Stranded Conformational; D012367:RNA, Viral / Q000302:isolation & purification; D014779:Virus Replication
D012367:RNA, Viral