[Cloning and antitumor activity of angiogenesis inhibitor HIAF-1].
OBJECTIVE
A new angiogenesis inhibitor HIAF-1 (human inhibitor angiogenesis factor-1) was cloned, expressed in E. coli and its antitumor activity was studied.


METHODS
HIAF-1 was amplified by RT-PCR from human fetal liver tissue, then cloned into pET30a(+) vector and expressed in E. coli BL21:DE3 after sequencing. In vitro endothelial proliferation inhibiting activity of HIAF-1 was examined by MTT method. In vivo antitumor activity was studied in a murine model of IVTA2MA-891.


RESULTS
HIAF-1 was first cloned from human fetal liver tissue. Sequence analysis of the inhibitor revealed identity to a c-terminal fragment of human collagen XVIII. HIAF-1 was effectively expressed in E. coli with a yield of 88 mg/L. Recombinant HIAF-1 protein could inhibit endothelial cell proliferation in vitro with an IC50 value of 7.5 micrograms/ml. In vivo studies showed that HIAF-1 inhibited growth rate of the primary tumor by 46.6% and metastasis by 68.9%.


CONCLUSIONS
The cloned and the bio-engineering product of HIAF-1 is an angiogenesis inhibitor, capable of inhibiting tumor growth and metastasis.
Guo
W
W
Cancer Institute (Hospital), Chinese Academy of Medical Science, Peking Union Medical College, Beijing 100021, China.
Yang
Z
Z
Liu
J
J
D020533:Angiogenesis Inhibitors / Q000235:genetics* / Q000494:pharmacology* / Q000627:therapeutic use; D000818:Animals; D001158:Arteries / Q000187:drug effects*; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D003001:Cloning, Molecular / Q000379:methods; D003094:Collagen / Q000235:genetics; D043170:Collagen Type XVIII; D004195:Disease Models, Animal; D043169:Endostatins; D004727:Endothelium / Q000187:drug effects; D015870:Gene Expression; D020128:Inhibitory Concentration 50; D051379:Mice; D009362:Neoplasm Metastasis / Q000517:prevention & control; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000188:drug therapy / Q000517:prevention & control; D009389:Neovascularization, Pathologic / Q000188:drug therapy / Q000517:prevention & control; D010446:Peptide Fragments / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D017385:Sequence Homology; D023041:Xenograft Model Antitumor Assays
D020533:Angiogenesis Inhibitors; D043170:Collagen Type XVIII; D043169:Endostatins; C448783:HIAF-1 protein, human; D010446:Peptide Fragments; D011506:Proteins; D011994:Recombinant Proteins; D003094:Collagen