Dynamic distribution and expression in vivo of human endostatin gene delivered by adenoviral vector.
Endostatin, a 20-kDa carboxyl-terminal fragment of collagen XVIII, is a potent inhibitor of endothelial cell proliferation and tumor angiogenesis. We have constructed replication-deficient recombinant adenovirus (Ad-rhE), which encoded secreted human endostatin, and our previous studies showed that Ad-rhE had a potent suppression of tumor growth in vivo. In the present study, we investigated the dynamic distribution and expression of human endostatin gene in vivo using fluorogenic real-time quantitative PCR and enzyme-linked immunosorbent assay(ELISA), respectively, with an injection of 2.0 x10(9)pfu of Ad-rhE. After injection, the Ad-rhE DNAs decreased sharply, but lasted a relative long-term at low concentration (10,000--20,000 copies/mg tissues). Whereas the expressed endostatin rose up rapidly, and reached to the top on day 5 after injection of Ad-rhE, and then decreased sharply, but endostatin in tumors sustained to over 9 days at a certain level. Both Ad-rhE DNAs and endostatin mainly enriched in tumors in vivo, and then in livers. These results suggest that endostatin gene delivered by adenoviral vector can generate a high expression in vivo, and both the metabolism pathways of Ad-rhE DNAs and endostatin in vivo are through the systems of livers.
He
Guo-An
GA
State Key Laboratory of Tumors, Cancer Center, Sun Yat-sen University, Guangzhou 510060, PR China.
Xue
Gang
G
Xiao
Lin
L
Wu
Jiang-Xue
JX
Xu
Ben-Ling
BL
Huang
Jia-Ling
JL
Liang
Zhi-Hui
ZH
Xiao
Xia
X
Huang
Bi-Jun
BJ
Liu
Ran-Yi
RY
Huang
Wenlin
W
D000256:Adenoviridae / Q000235:genetics*; D045744:Cell Line, Tumor; D043169:Endostatins / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D007267:Injections; D008099:Liver / Q000378:metabolism; D008546:Melanoma, Experimental / Q000378:metabolism; D009368:Neoplasm Transplantation; D020133:Reverse Transcriptase Polymerase Chain Reaction
D043169:Endostatins