[Inhibitory effect of CVB3-VP1 siRNA on CVB3 replication].
OBJECTIVE
To inhibit the expression of CVB3 VP1 protein and the replication of CVB3 with synthesized siRNAs.


METHODS
According to the sequence and secondary structure of CVB3 VP1 protein, four pieces of siRNAs were designed following the requirement from Journal of Nature Cell Biology were synthesized in Shanghai GeneChem Company. Then they were transfected into HeLa cells by liposome (Lipofectamine 2000), but the non-transfected cells and non-specific siRNAs were taken as control. 48 hours later, the patho-morphous changes were observed, virus titer changes were examined by TCID50, CVB3-VP1 protein expression were detected by immunofluorescence with FITC dyeing, and CVB3-RNA level was tested by semi-quantitative RT-PCR.


RESULTS
Two pieces of the four specific synthesized siRNAs (VP1-1 and VP1-2) were found to have obvious inhibitory effect on CVB3 replication and VP1 protein expression were reduced greatly. Besides, the changes of pathological cells were obviously mitigated.


CONCLUSIONS
Specific siRNAs can effectively inhibit the expression of CVB3 VP1 protein and the replication of CVB3 in HeLa cells.
Chen
Jie-bin
JB
Department of Pediatrics, the First Affiliated Hospital, Nanhua University, Hengyang 421001, China. cjb129850716@163.com
Xu
Cong-feng
CF
Yu
Yong
Y
Li
Li
L
Li
Shuang-jie
SJ
Chen
Rui-zhen
RZ
Yang
Ying-zhen
YZ
D002681:China; D004261:DNA Replication / Q000187:drug effects; D029822:Enterovirus B, Human / Q000187:drug effects* / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D034622:RNA Interference / Q000187:drug effects; D034741:RNA, Small Interfering / Q000494:pharmacology*; D014779:Virus Replication / Q000187:drug effects*
D034741:RNA, Small Interfering