[Transfer and expression of antisense genes of hepatitis B virus (HBV) and their anti-HBV effects].
OBJECTIVE
To observe the transfer and expression of HBV antisense genes and their anti-HBV effects in 2.2.15 cells mediated by retroviral vector-packaging cell line system.


METHODS
The recombinant retroviral vectors mediating expression of HBV antisense RNA complementary to 1402-2906 fragment (preC/C) or 2839-1986 fragment (preS/S) of HBV DNA were used to transduce into NIH 3T3 cells and 2.2.15, respectively. Then, the total RNAs of tranduced NIH 3T3 cells were extracted and hybridized with HBV DNA probe. The HBsAg and HBeAg in the supernatants of cultured 2.2.15 cells were assayed with RIA method and the DNAs were hybridized with HBV DNA probe by dot blot.


RESULTS
The results showed that the HBV antisense genes could be transfered into and expressed in the NIH 3T3 cells mediated by recombinant retroviral vector packaging cell line (PA317 cell). The inhibitory effects on the expression of HBV antigens by antisense RNAs appeared as early as on the day 3 after transduced, reached peak level on the day 5, and persisted at least for eleven days. The inhibitory rates of HBsAg and HBeAg were 71% and 23% by antisense preS/S, and 23% and 59% by antisense preC/C on the day 5 after transduction. In comparison to blank control or sense--gene expressing vectors, the inhibitory effects of HBV antisense vectors were highly significant (P < 0.01). HBV DNA level in the supernatant of the 2.2.15 cells transduced with either antisense preS/S or preC/C in comparison to the blank control vectors was also reduced on the day 5 after transduction as detected by DNA dot blot assays, but the viability of transduced 2.2.15 cells was not affected as detected by MTT assays.


CONCLUSIONS
HBV antisense genes can be transfered and expressed in the eukaryotic cells and the expression of HBV antisense RNAs in 2.2.15 cells can inhibit the replication and expression of HBV.
Department of Infectious Diseases, First Affiliated Hospital, Beijing Medical University.
W
W
Ji
Q
Q
Wang
M
M
Yu
D016475:3T3 Cells; D000818:Animals; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D051379:Mice; D016372:RNA, Antisense / Q000096:biosynthesis*; D012367:RNA, Viral / Q000096:biosynthesis*; D014162:Transfection
D016372:RNA, Antisense; D012367:RNA, Viral