[Transfection of siRNA expressing plasmids targeting S gene inhibits replication and expression of hepatitis B virus in hepatic cancer cells].
OBJECTIVE
To study the inhibitive effects of transfection of siRNA expressing plasmids targeting S gene, one of the 4 open reading frames of hepatitis B virus (HBV), on the replication and expression of HBV.


METHODS
Two plasmids expressing 2 siRNAs targeting S gene, one of the 4 open reading frames of HBV (S1 and S2) and one nonspecific plasmid (siRNA-S3), as negative control, with the length of 21 nt heterologous to the HBV/U95551 genome were constructed, and then transfected into the hepatic cancer cells of the line HepG2.2.15. 48 hours later, real-time PCR was used to evaluate the gene silencing efficiency and ELISA was used to detect the expression of HBsAg and hepatitis B e antigen (HBeAg), protein markers of HBV, in the supernatants.


RESULTS
The inhibition rates of HBsAg and HBeAg expression of the HepG2.2.15 cells transfected with S1 were 60% and 56% respectively, those of the HepG2.2.15 cells transfected with S2 were 73% and 70% respectively, those of the HepG2.2.15 cells transfected with S1+S2 were 82% and 78% respectively, and those of the HepG2.2.15 cells transfected with S3 were not significantly different from those of the blank control group. RT-PCR showed that the mRNA expression rates of HBsAg and HBeAg in the HepG2.2.15 cells transfected with S1, S2, and S1+S2 were inhibited by 64%-88% t respectively.


CONCLUSIONS
Transfection of the vector plasmids expressing the siRNAs targeting S gene inhibits the expression of HBsAg and HBeAg in hepatic cancer cells. RNAi may provide a viable strategy against viruses for the prevention and treatment of HBV infection.
Bian
Zhong-qi
ZQ
Center for Infectious Diseases, Kunming General Hospital of Chinese People's Liberation Army, Kunming 650032, China. bzq@ynu.edu.cn
Cui
Zhi-lei
ZL
Chen
Wei-zao
WZ
Liu
Ming-qiu
MQ
Yan
Wei-yao
WY
Zheng
Zhao-xin
ZX
D045744:Cell Line, Tumor; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D016366:Open Reading Frames; D034622:RNA Interference; D034741:RNA, Small Interfering; D012367:RNA, Viral; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics*; D014779:Virus Replication
D034741:RNA, Small Interfering; D012367:RNA, Viral; C118669:S envelope protein, hepatitis B virus; D014759:Viral Envelope Proteins