Expression and purification of human endostatin in Pichia pastoris and its inhibition on the growth of mouse pulmonary adenocarcinoma cell line LA795.
OBJECTIVE
To obtain yeast strain Pichia pastoris that highly expresses human endostatin by gene recombination technology, and to evaluate the inhibitory effect of recombinant human endostatin (rhES) on the growth of mouse pulmonary adenocarcinoma cell line LA795.


METHODS
The gene coding for human endostatin was cloned into the genome of Pichia pastoris through LiCl transformation method, and the clones with high soluble rhES expression were selected. Purification of rhES was performed with heparin affinity chromatography. Effects of rhES on bFGF-induced proliferation of human endothelial cell line ECV-304 cells were observed. T739 mice with subcutaneous inoculation of LA795 cells were treated with injections of either rhES or PBS for 14 consecutive days, and the volume of the tumors were measured.


RESULTS
Clones with high rhES expression were obtained and purified rhES potently inhibited the proliferation of ECV-304 cells and the growth of LA795 cells in T739 mice.


CONCLUSIONS
rhES produced by Pichia pastoris possesses good biological activities and conspicuously inhibits the growth of LA795 cells.
Xia
Hu
H
Department of Respiratory Diseases, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China.
Zhang
Lin
L
Wen
Jin-Xu
JX
Tong
Wan-Cheng
WC
D000230:Adenocarcinoma / Q000188:drug therapy* / Q000473:pathology; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D003094:Collagen / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D043169:Endostatins; D015870:Gene Expression; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy* / Q000473:pathology; D008297:Male; D051379:Mice; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D043169:Endostatins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D003094:Collagen