BC047440 antisense eukaryotic expression vectors inhibited HepG2 cell proliferation and suppressed xenograft tumorigenicity

The biological functions of the BC047440 gene highly expressed by hepatocellular carcinoma (HCC) are unknown. The objective of this study was to reconstruct antisense eukaryotic expression vectors of the gene for inhibiting HepG2 cell proliferation and suppressing their xenograft tumorigenicity. The full-length BC047440 cDNA was cloned from human primary HCC by RT-PCR. BC047440 gene fragments were ligated with pMD18-T simple vectors and subsequent pcDNA3.1(+) plasmids to construct the recombinant antisense eukaryotic vector pcDNA3.1(+)BC047440AS. The endogenous BC047440 mRNA abundance in target gene-transfected, vector-transfected and naive HepG2 cells was semiquantitatively analyzed by RT-PCR and cell proliferation was measured by the MTT assay. Cell cycle distribution and apoptosis were profiled by flow cytometry. The in vivo xenograft experiment was performed on nude mice to examine the effects of antisense vector on tumorigenicity. BC047440 cDNA fragments were reversely inserted into pcDNA3.1(+) plasmids. The antisense vector significantly reduced the endogenous BC047440 mRNA abundance by 41 percent in HepG2 cells and inhibited their proliferation in vitro (P < 0.01). More cells were arrested by the antisense vector at the G1 phase in an apoptosis-independent manner (P = 0.014). Additionally, transfection with pcDNA3.1(+)BC047440AS significantly reduced the xenograft tumorigenicity in nude mice. As a novel cell cycle regulator associated with HCC, the BC047440 gene was involved in cell proliferation in vitro and xenograft tumorigenicity in vivo through apoptosis-independent mechanisms.

A study of mechanism of modulation of NF-κB activity of HepG2 cells by BC047440 genes / 中华肝胆外科杂志

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.

BC047440 gene silencing inhibits the proliferation of HepG2 cells / 中华消化外科杂志

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.

Expression and Significance of A Novel Gene BC047440 in Multiplicate Malignant Tumor Tissues / 中国普外基础与临床杂志

0.05).②There were close relationships between BC047440 gene expression and clinicopathologic findings of liver cancer,including tumor size and portal vein invasion(P

Expression of BC047440 gene in human hepatocellular carcinoma tissues and its significance / 中国普通外科杂志

Objective To study the BC047440 mRNA expression in human hepatocellular carcinoma tissues, and the role of BC047440 gene in the carcinogenesis and development of human hepatocellular carcinoma.Methods Specimens from 36 cases of hepatocellular carcinoma and their corresponding adjacent non-cancerous tissues were examined for BC047440 mRNA expression by polymerase chain reaction after reverse transcription(RT-PCR).Results(1)the BC047440 mRNA expression in specimens of 36 cases of hepatocellular carcinoma was higher than that in their adjacent non-cancerous tissues(0.2594?0.0928 and 0.0942?0.0443,respectively,P