AIM： To investigate the anti-angiogenic and antitumor activities of recombinant vascular basement membrane-derived multifunctional peptide （rVBMDMP） in hepatocellular carcinoma （HCC）. METHODS： HepG2, Bel-7402, H...AIM： To investigate the anti-angiogenic and antitumor activities of recombinant vascular basement membrane-derived multifunctional peptide （rVBMDMP） in hepatocellular carcinoma （HCC）. METHODS： 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 ICs0 was 4.68, 7.65, 8.96, 11.65 and 64.82 μmol/L, respectively, the potency of rVBMDMP to HepG2 cells was similar to 5-fluorouracil （5-FU） with an IC50 of 4.59 μmol/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%. AIternis 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： rVBMDMP has effective and unique anti-tumor properties, and is a promising candidate for the development of anti-tumor drugs.展开更多
基金Supported by The Nation Natural Science Foundation of China, No. 30472040the Key Program of the Health Department of Hunan Province, No. 2004-005the National Undergraduate Innovative Test Program, No. YA07059 and No. 081054239
文摘AIM： To investigate the anti-angiogenic and antitumor activities of recombinant vascular basement membrane-derived multifunctional peptide （rVBMDMP） in hepatocellular carcinoma （HCC）. METHODS： 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 ICs0 was 4.68, 7.65, 8.96, 11.65 and 64.82 μmol/L, respectively, the potency of rVBMDMP to HepG2 cells was similar to 5-fluorouracil （5-FU） with an IC50 of 4.59 μmol/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%. AIternis 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： rVBMDMP has effective and unique anti-tumor properties, and is a promising candidate for the development of anti-tumor drugs.
