Combination of small interfering RNA and lamivudine on inhibition of human B virus replication in HepG2.2.15 cells.
OBJECTIVE
To observe the inhibition of hepatitis B virus (HBV) replication and expression by combination of siRNA and lamivudine in HepG2.2.15 cells.


METHODS
Recombinant plasmid psil-HBV was constructed and transfected into HepG2.2.15 cells. The transfected cells were cultured in lamivudine-containing medium (0.05 micromol/L) and harvested at 48, 72 and 96 h. The concentration of HBeAg and HBsAg was determined using ELISA. HBV DNA replication was examined by real-time PCR and the level of HBV mRNA was measured by RT-PCR.


RESULTS
In HepG2.2.15 cells treated with combination of siRNA and lamivudine, the secretion of HBeAg and HBsAg into the supernatant was found to be inhibited by 91.80% and 82.40% (2.89+/-0.48 vs 11.73+/-0.38, P<0.05; 4.59+/-0.57 vs 16.25+/-0.48, P<0.05) at 96 h, respectively; the number of HBV DNA copies within culture medium was also significantly decreased at 96 h (1.04+/-0.26 vs 8.35+/-0.33, P<0.05). Moreover, mRNA concentration in HepG2.2.15 cells treated with combination of siRNA and lamivudine was obviously lower compared to those treated either with siRNA or lamivudine (19.44+/-0.17 vs 33.27+/-0.21 or 79.9+/-0.13, P<0.05).


CONCLUSIONS
Combination of siRNA and lamivudine is more effective in inhibiting HBV replication as compared to the single use of siRNA or lamivudine in HepG2.2.15 cells.
Li
Gui-Qiu
GQ
Department of Microbiology, Harbin Medical University, Harbin 150081, and Department of Anesthesiology, Second Affiliated Hospital, Jiamusi University, Heilongjiang Province, China.
Xu
Wei-Zhen
WZ
Wang
Jing-Xia
JX
Deng
Wen-Wei
WW
Li
Di
D
Gu
Hong-Xi
HX
D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use; D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000821:virology; D045744:Cell Line, Tumor; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000378:metabolism; D004359:Drug Therapy, Combination; D006509:Hepatitis B / Q000188:drug therapy; D006514:Hepatitis B Surface Antigens / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology* / Q000627:therapeutic use; D008113:Liver Neoplasms / Q000473:pathology / Q000821:virology; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology* / Q000627:therapeutic use; D012367:RNA, Viral / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D019259:Lamivudine