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

Abstract

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% 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.

Bioinformatics Analysis and Insights for the Role of COMMD7 in Hepatocellular Carcinoma

“…Subsequently, a self-prepared monoclonal antibody to COMMD7 was uesd to detect the protein level of COMMD7 in 7 liver cancer cell lines including HepG2, Huh7, Hep3B, HLE, HLF, SK-Hep-1 and PLC/PRF5. The results revealed that COMMD7 protein expressed totally higher in these hepatoma cells than in normal hepatocytes, especially in HepG2 and SK-Hep-1 [36,37]. For further study, we constructed a pGenesil-COMMD7-shRNA plasmid both in HepG2 and SK-Hep-1cells by means of short-hairpin RNA targeted technology.…”

Proliferation inhibition and differentiation induction of hepatic cancer stem cells by knockdown of BC047440: A potential therapeutic target of stem cell treatment for hepatocellular carcinoma

“…The current studies regarding the roles of BC047440, a new HCC-related gene, in HCC were all completed by our research group, and the results have been widely recognized (7)(8)(9). In a previous investigation, we found that downregulated BC047440 expression can inhibit HepG2 cell proliferation, decreasing cellular AFP expression and significantly increasing functional gene expression in normal hepatocytes, such as ALB and cytochrome P450 1A2 (CYP1A2) (Fig.…”

“…Furthermore, evidence suggests that BC047440 plays a crucial role in mediating HCC proliferation and differentiation ( Fig. 1) of HCC cells (7)(8)(9). Nevertheless, few studies have assessed the role of BC047440 in HCSCs.…”