Secreted human apolipoprotein(a) kringle IV-10 and kringle V inhibit angiogenesis and xenografted tumor growth.
Abstract Angiogenesis plays an important role in normal physiology of blood vessel growth, but can contribute to the pathogenesis of diseases, such as cancer. A new anti-angiogenic recombinant kringle protein, composed of the fused domains of human apolipoprotein(a) carboxyl-terminal kringle IV-10 and kringle V, was expressed in Pichia pastoris and human colorectal carcinoma (HCT 116) cells to investigate its influence on angiogenesis and tumor growth. The mature recombinant protein exhibited the characteristic features of kringle-containing proteins (glycosylation and disulfide bond formation) and, when added to cultures of human umbilical vein endothelial cell, resulted in a 31% decrease in proliferation relative to untreated controls (p<0.05). The neo-angiogenesis was diminished by 63% in chick embryos treated with 10 mug recombinant protein compared with 7% for phosphate buffer solution-treated embryos (p<0.01). Transfection of a kringle IV-10-kringle V fusion protein construct into HCT 116 cells decreased tumorigenesis and inhibited tumor growth in vivo without affecting tumor cell proliferation. HCT 116 cells that expressed recombinant protein displayed a much lower relative growth ratio of 8% (p<0.01) against the control tumor cells. From these results, we conclude that human apolipoprotein(a) carboxyl-terminal kringle IV-10-kringle V fusion protein is an effective inhibitor of angiogenesis and angiogenesis-dependent tumor growth.
Shen
Le
L
Department of Biochemistry and Molecular Biology, National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing 100005, PR China.
Zhu
Xuewei
X
Wang
Yan
Y
Zeng
Wuwei
W
Wu
Gang
G
Xue
Hong
H
Chen
Baosheng
B
D020533:Angiogenesis Inhibitors / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D000818:Animals; D000970:Antineoplastic Agents / Q000627:therapeutic use*; D001054:Apolipoproteins A / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D045744:Cell Line, Tumor; D002642:Chick Embryo; D003001:Cloning, Molecular; D015179:Colorectal Neoplasms / Q000473:pathology; D006801:Humans; D018082:Kringles / Q000235:genetics* / Q000502:physiology; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000188:drug therapy* / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000517:prevention & control; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use; D014183:Transplantation, Heterologous
D020533:Angiogenesis Inhibitors; D000970:Antineoplastic Agents; D001054:Apolipoproteins A; D011993:Recombinant Fusion Proteins