[High expression of human angiogenesis inhibitor HIAF-1 in insect cells and biological activity].
OBJECTIVE
Human inhibiting angiogenesis factor-1(HIAF-1) expressed in E coli. shown the activity of inhibiting the angiogenesis of tumors. This study was designed to investigate the expression of HIAF-1 in insect cells and to elucidate its biological activity.


METHODS
Recombinant baculovirus expression vector pAcuW51-HIAF-1 was constructed by recombinant DNA technique and cotransfected with linearized baculovirus DNA into sf9 cells to produce recombinant virus. The expression of recombinant HIAF-1 in insect cells infected by recombinant virus was detected by SDS-PAGE and Western blot. The recombinant protein was purified by affinity chromatography. In vitro endothelial proliferation inhibiting activity of recombinant HIAF-1 was examined by MTT method, its antitumor activity in vivo was studied in human esophageal cancer transplanted in nude mice.


RESULTS
HIAF-1 was effectively expressed in insect cells as 26 KD fusing protein and its expression level was about 5-10% of insect cellular total soluble proteins. Recombinant HIAF-1 protein could inhibit endothelial cell proliferation in vitro with IC50 value of 3.1 micrograms/ml and inhibited remarkably growth of human esophageal cancer transplanted in nude mice.


CONCLUSIONS
The recombinant HIAF-1 with better activity was successfully expressed in insect cells and establish a base for clinical application.
Guo
Wen-zhong
WZ
Department of Cellular Biology and Molecular Biology, Cancer Institute, Peiking Union Medical College, Beijing 100021, P. R. China.
Ran
Yu-liang
YL
Liu
Jun
J
Yu
Long
L
Sun
Li-xin
LX
Yang
Zhi-hua
ZH
D020533:Angiogenesis Inhibitors / Q000096:biosynthesis* / Q000494:pharmacology; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D004727:Endothelium / Q000187:drug effects; D015870:Gene Expression; D006801:Humans; D007313:Insecta / Q000166:cytology; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology
D020533:Angiogenesis Inhibitors; C448783:HIAF-1 protein, human; D011506:Proteins; D011994:Recombinant Proteins