Huang Yi,Yang Meihua,Yang Huaan,Jiang Xue,Zeng Zhaochun.Inhibitory effect of silencing GW112 siRNA gene on proliferation of human gastric SGC-7901 cell line[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(13):1379-1382.
Huang Yi,Yang Meihua,Yang Huaan,Jiang Xue,Zeng Zhaochun.Inhibitory effect of silencing GW112 siRNA gene on proliferation of human gastric SGC-7901 cell line[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(13):1379-1382.DOI:
Inhibitory effect of silencing GW112 siRNA gene on proliferation of human gastric SGC-7901 cell line
Objective To construct the small interfering RNA (siRNA) retrovirus vector targeting GW112 with a high specificity and to study the effect of silencing GW112 gene on the proliferation of gastric SGC-7901 cells. Methods Three recombinant RNAi interference vectors were constructed according to different GW112 targeting points (pSi405-GW112
pSi1066-GW112
pSi1480-GW112) using a retrovirus expression vector
psilencer 5.1-H1 Retro. SGC- 7901 cells were infected with a packaged PT67 cells transfected with liposome. Stable clones were selected using puromycin. Integration of virus was identified by RT-PCR and highly inhibited targets were detected RT-PCR. The effect of GW112 on proliferation of gastric SGC-7901 cells was detected with CCK-8 kit. Expression of proliferating cell nuclear antigen (PCNA) was assayed by Western blotting. Results Sequence analysis showed recombinant retrovirus vector psiRNA-GW112 was successfully constructed. RT-PCR showed that except for parental cells
other SGC-7901 cells infected with virus supernatant could amplify anti-puromycin GW112 gene into a 510 bp fragment. The inhibitory efficacy was 84.5%,67.6%
and 43.0%
respectively
for Si405
Si1066 and Si1480 respectively. Both the proliferation of gastric SGC-7901 cells and the expression of PCNA in SGC-7901 cells were significantly inhibited after GW112 was silenced. Conclusion The recombinant retrovirus vector
pSiRNA-GW112
can significantly inhibit the proliferation and expression of PCNA in SGC-7901 cells
indicating that GW112 plays an important role in proliferation of SGC-7901cells.