Published online Sep 14, 2018. doi: 10.3748/wjg.v24.i34.3861
Peer-review started: May 25, 2018
First decision: June 21, 2018
Revised: July 5, 2018
Accepted: July 16, 2018
Article in press: July 16, 2018
Published online: September 14, 2018
To investigate the role of Delta-like ligand 4 (DLL4) on tumour growth in hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) in vivo.
We suppressed DLL4 expression in an HBV expressing HCC cell line, HepG2.2.15 and analysed the growth ability of cells as subcutaneous tumours in nude mice. The expression of tumour angiogenesis regulators, VEGF-A and VEGF-R2 in tumour xenografts were examined by western blotting. The tumour proliferation and neovasculature were examined by immunohistochemistry. The viral replication and viral protein expression were measured by quantitative PCR and western blotting, respectively.
Eighteen days after implantation, tumour volume in mice implanted with shDLL4 HepG2.2.15 was significantly smaller than in mice implanted with control HepG2.2.15 (P < 0.0001). The levels of angiogenesis regulators, VEGF-A and VEGF-R2 were significantly decreased in implanted tumours with suppressed DLL4 compared with the control group (P < 0.001 and P < 0.05, respectively). Furthermore, the suppression of DLL4 expression in tumour cells reduced cell proliferation and the formation of new blood vessels in tumours. Unexpectedly, increased viral replication was observed after suppression of DLL4 in the tumours.
This study demonstrates that DLL4 is important in regulating the tumour growth of HBV-associated HCC as well as the neovascularization and suppression of HBV replication.
Core tip: We demonstrated that Delta-like ligand 4 (DLL4) is important for tumour growth of hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC) in a xenograft model. We found that the level of angiogenesis regulators, VEGF-A and VEGF-R2 were significantly decreased in HCC xenograft tumours with suppressed DLL4 compared with the control group. Consistent with these findings, the suppression of DLL4 expression in the tumour cells reduced cell proliferation and the formation of new blood vessels in the tumour. Furthermore, this is the first report that DLL4 in an HBV expressing HCC cell line plays a key role in regulating tumour growth, angiogenesis, and viral replication in a mouse model of xenograft transplantation.