HepG2, Bel-7402, Hep-3B, HUVE-12 and L-02 cell lines were cultured in vitro and the inhibitory effect of
rVBMDMP on proliferation of cells was detected by MTT assay. The in vivo antitumor efficacy of
rVBMDMP on HCC was assessed by HepG2 xenografts in nude mice. Distribution of
rVBMDMP, mechanism by which the growth of HepG2 xenografts is inhibited, and microvessel area were observed by
proliferating cell nuclear antigen (
PCNA) and CD31 immunohistochemistry.
RESULTS: MTT assay showed that
rVBMDMP markedly inhibited the proliferation of human HCC (HepG2, Bel-7402, Hep-3B) cells and human umbilical vein endothelial (HUVE-12) cells in a dose-dependent manner, with little effect on the growth of L-02 cells. When the IC(50) was 4.68, 7.65, 8.96, 11.65 and 64.82 micromol/L, respectively, the potency of
rVBMDMP to HepG2 cells was similar to
5-fluorouracil (5-FU) with an IC(50) of 4.59 micromol/L. The selective index of cytotoxicity to HepG2 cells of
rVBMDMP was 13.8 (64.82/4.68), which was higher than that of
5-FU [SI was 1.9 (8.94/4.59)]. The
VEGF-targeted recombinant humanized
monoclonal antibody bevacizumab (100 mg/L) did not affect the proliferation of HepG2, Bel-7402, Hep-3B and L-02 cells, but the growth inhibitory rate of
bevacizumab (100 mg/L) to HUVE-12 cells was 87.6% +/- 8.2%. Alternis diebus
intraperitoneal injection of
rVBMDMP suppressed the growth of HepG2 xenografts in a dose-dependent manner.
rVBMDMP (1, 3, 10 mg/kg) decreased the
tumor weight by 12.6%, 55.9% and 79.7%, respectively, compared with the vehicle control. Immunohistochemical staining of
rVBMDMP showed that the positive area rates (2.2% +/- 0.73%, 4.5% +/- 1.3% and 11.5% +/- 3.8%) in
rVBMDMP treated group (1, 3, 10 mg/kg) were significantly higher than that (0.13% +/- 0.04%) in the control group (P < 0.01). The positive area rates (19.0% +/- 5.7%, 12.2% +/- 3.5% and 5.2% +/- 1.6% ) of
PCNA in
rVBMDMP treated group (1, 3, 10 mg/kg) were significantly lower than that (29.5% +/- 9.4%) in the control group (P < 0.05).
rVBMDMP at doses of 1, 3 and 10 mg/kg significantly reduced the
tumor microvessel area levels (0.26% +/- 0.07%, 0.12% +/- 0.03% and 0.05% +/- 0.01% vs 0.45% +/- 0.15%) in HepG2 xenografts (P < 0.01), as assessed by CD31 staining.
CONCLUSION: