title: Effects of short RNA structural analogues against hepatitis C virus genotypes 2, 3 and 4 in replicon cells.
abstract: OBJECTIVE
  To determine whether computer-predicted short RNA structural analogues could inhibit hepatitis C virus (HCV) genotype 2a, 3a and 4a replication in cultured cells.
  
  
  METHODS
  Short RNA sequences, X12, X12a and X12b, designed to be identical in secondary structure to the X region in the 3'-untranslated region (3'-UTR) of the HCV 1b genome, as well as shorter stem-loop components of X region, were inserted into a plasmid and transfected into separate Huh7.5 human hepatoma cells stably transfected with subgenomic replicons for genotypes 2a, 3a and 4a. All replicons included a firefly luciferase reporter gene. After 48 h of plasmid transfection, the inhibition of HCV replication was determined by HCV RNA isolation and quantification by real-time polymerase chain reaction and luciferase assays.
  
  
  RESULTS
  All the secondary structural analogues to genotype 1b X region cross-inhibited genotype 2a, 3a and 4a replicons. The maximum inhibition by genotype 1b X region structural analogues was obtained against genotype 2a cells in which X12, X12a and X12b inhibited replication by 30%, 63% and 72%, respectively (P < 0.05 for all), compared to an unrelated hepatitis B viral analogue.
  
  
  CONCLUSIONS
  Despite substantial sequence dissimilarity, HCV RNA genotype 1b X region analogues cross-inhibited the replication of HCV genotypes 2a, 3a and 4a. Particular conformations and not the sequence of the stem-loops of the X region are involved in HCV replication.
authors: Department of Biochemistry, Faculty of Pharmacy, Misr International University, Cairo, Egypt.
authors: Ismail M
authors: IM
authors: Elshaffei
authors: Department of Medicine, Division of Gastroenterology-Hepatology, UCONN HEALTH, USA.
authors: Nidhi
authors: N
authors: Gupta
authors: Department of Medicine, Division of Gastroenterology-Hepatology, UCONN HEALTH, USA.
authors: Catherine H
authors: CH
authors: Wu
authors: Department of Medicine, Division of Gastroenterology-Hepatology, UCONN HEALTH, USA.
authors: David C
authors: DC
authors: Wu
authors: Department of Biochemistry, Faculty of Pharmacy, Misr International University, Cairo, Egypt.
authors: Lamiaa N
authors: LN
authors: Hammad
authors: Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
authors: Dina M
authors: DM
authors: Abo-Elmatty
authors: Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
authors: Noha M
authors: NM
authors: Mesbah
authors: Department of Medicine, Division of Gastroenterology-Hepatology, UCONN HEALTH, USA.
authors: George Y
authors: GY
authors: Wu
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000821:virology; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008113:Liver Neoplasms / Q000821:virology; D008156:Luciferases / Q000378:metabolism; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D012093:Replicon / Q000235:genetics*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014779:Virus Replication / Q000235:genetics
chemical_list: D012367:RNA, Viral; D008156:Luciferases
keywords: RNA structural analogue; cross inhibition; hepatitis C virus; replication