| Objectives The study research SiRNA silence Fra-1gene expression in human gastriccancer cell MGC-803，and further observe the effect of COX-2, NF-κB expression,and explore the possible mechanism of Fra-1gene regulation of gastric cancer cellprol-iferation.Methods This experiment was divided into4groups：Fra-1SiRNA intervention group，control SiRNA intervention group，empty liposome group and blankcontrol group.Fra-1SiRNA was transferred into MGC-803cell-s by lipofection2000，then observed its transfection efficiency by fluorescence microscope.Cellproliferation inhibition was measured at24，48and72h after transfection byCCK-8.The change in cells apoptosis rate was detected at48h after transferction by flow cytometry. the expressions of mRNA Fra-1,NF-κB,COX-2were tested byRT-PCR; the expressions of protein Fra-1,NF-κB,COX-2were determinedby Western-Blot assay.Results1.Transfection rate results:After48h by transfer Fra-1SiRNA，we can see green fluorescent in the cytoplasm，and the transfection efficiency canbe up to70%，the remaining three groups were no green fiuorescene.2.The profiferation inhibition rate achieved in the Fra-1SiRNA inter-vention group was higher than other groups. SiRNA transfection group can promoteapoptosis of human gastric cancer (P <0.05), the other three groups can not（P﹥0.05）.3.The expression of Fra-1,Fra-1mRNA and protein expression in the transfection SiRNA group was lower than other three groups(P﹤0.05), whereas nosignificant difference between the other three groups（P﹥0.05）. 4. The expression of NF-κB,COX-2,NF-κB and COX-2mRNA and protein expression in the transfection SiRNA group was lower than other three groups(P﹤0.05), whereas no signi-ficant difference between the other three groups（P﹥0.05）.Conclusions1The transfection of targeted Fra-1SiRNA into human gastric cancerMGC-803cells can result in silence of expression of Fra-1.2. Silence Fra-1can inhibit proliferation and promote apoptosis rate in gastriccancer cells of MGC-803, suggesting that the mechanism may be reduced NF-κB andCOX-2factor expression by Fra-1/NF-κB/COX-2pathway. |
