ABSTRACT
Objective To investigate the mechanism that BC047440 gene regulates nuclear fac-tor κB sigal passway and analyze the differential expression gene between HepG2 cells and HepG2 cells BC047440 gene silenced by RNAi using 35K Human Genome Array. Methods The differential expres-sion gene between HepG2 cells and HepG2 cells with BC047440 gene silenced was analyzed by 35K Human Genome Array, and the data were submitted to the database and MAS system of Capitalbio Corporation.Then TRAF6 was confirmed by RT-PCR test. Results Among the total 35000 probe sets, the expression of 59 genes was down-regulated for more than 50% and 130 genes were up-regulated more than 2 fold in the silencing group when compared with normal controls. TRAF6 mRNA was decreased for 29.5% in silicening HepG2 compared with that of wild HepG2 by RT-PCR, which is similar to human genome array(23.06%).Conclusion The high throughput and effective oligomicroarray can analyze the differential expression gene and BC047440 gene might regulate NF-κB signal pathway inderectly by TRAF6.
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Objective To investigate whether the proliferation of HepG2 ceils is influenced by interfering BC047440 gene with small hairpin RNA (shRNA) expression plasmid. Methods According to the sequence of BC047440 gene, 2 pairs of BC047440 gene-specific shRNA (shRNA1 and shRNA2) were designed and synthesized. After primer annealing, they were inserted into plasmid pGenesil-1 to construct the shRNA expression plasmids. The recombinant plasmids were transfected into HepG2 cells. The expression of BC047440 gene was detected by quantitative fluorescent PCR, the proliferation of HepG2 cells by MTT assay and the changes of cell cycle by flow cytometry. Results Two shRNA expression plasmids were constructed successfully and were confirmed by restriction enzyme digestion and sequencing. Quantitative fluorescent PCR analysis showed that shRNA1 and shRNA2 could specifically inhibit the expression of BC047440 gene in HepG2 cells, with the inhibition rate of 80.22% and 58.63%, respectively. The shRNA effectively inhibited the proliferation of HepG2 cells, and arrested the cell cycle in S phase. Conclusions The shRNA significantly inhibits the expression of BC047440 gene and the proliferation of HepG2 cells. The expression of BC047440 may be correlated with the proliferation of HepG2 cells.
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0.05).②There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer,including tumor size and portal vein invasion(P