Document Detail

Expression of angiostatin cDNA in human gallbladder carcinoma cell line GBC-SD and its effect on endothelial proliferation and growth.
MedLine Citation:
PMID:  16718765     Owner:  NLM     Status:  MEDLINE    
AIM: To explore the influence of angiostatin up-regulation on the biologic behavior of gallbladder carcinoma cells in vitro and in vitro, and the potential value of angiostatin gene therapy for gallbladder carcinoma.
METHODS: A eukaryotic expression vector of pcDNA3.1(+) containing murine angiostatin was constructed and identified by restriction endonuclease digestion and sequencing. The recombinant vector pcDNA3.1-angiostatin was transfected into human gallbladder carcinoma cell line GBC-SD with Lipofectamine 2000, and paralleled with the vector and mock control. The resistant clone was screened by G418 filtration. Angiostatin transcription and protein expression were examined by RT-PCR, immunofluorescence and Western-blot. The supernatant was collected to treat endothelial cells. Cell proliferation and growth in vitro were observed under microscope.
RESULTS: Murine angiostatin cDNA was successfully cloned into the eukaryotic expression vector pcDNA3.1 (+). After 14 d of transfection and selection with G418, macroscopic resistant cell cloning was formed in the experimental group transfected with pcDNA 3.1(+)-angiostatin and vector control. But untreated cells died in the mock control. Angiostatin was detected by RT-PCR and protein expression was detected in the experimental group by immunofluorescence and Western-blot. Cell proliferation and growth in vitro in the three groups were observed respectively under microscope. No significant difference was observed in the growth speed of GBC-SD cells between groups that were transfected with and without angiostatin. After treatment with supernatant, significant differences were observed in endothelial cell (ECV-304) growth in vitro. The cell proliferation and growth were inhibited.
CONCLUSION: Angiostatin does not directly inhibit human gallbladder carcinoma cell proliferation and growth in vitro, but the secretion of angiostatin inhabits endothelial cell proliferation and growth.
Ding-Zhong Yang; Jing He; Ji-Cheng Zhang; Zuo-Ren Wang
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  World journal of gastroenterology : WJG     Volume:  12     ISSN:  1007-9327     ISO Abbreviation:  World J. Gastroenterol.     Publication Date:  2006 May 
Date Detail:
Created Date:  2006-05-23     Completed Date:  2006-07-03     Revised Date:  2014-09-09    
Medline Journal Info:
Nlm Unique ID:  100883448     Medline TA:  World J Gastroenterol     Country:  China    
Other Details:
Languages:  eng     Pagination:  2762-6     Citation Subset:  IM    
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MeSH Terms
Angiostatins / genetics*,  physiology
Blotting, Western
Carcinoma / chemistry,  genetics*,  pathology
Cell Line, Tumor
Cell Proliferation / drug effects*
Cell Survival / drug effects
Cells, Cultured
DNA, Complementary / analysis*,  genetics
DNA, Neoplasm / analysis*,  genetics
Endothelium, Vascular / cytology*,  growth & development
Gallbladder Neoplasms / chemistry,  genetics*,  pathology
Gene Expression Regulation, Neoplastic
Genetic Therapy
Genetic Vectors / genetics
Microscopy, Fluorescence
Neovascularization, Pathologic
Reverse Transcriptase Polymerase Chain Reaction
Reg. No./Substance:
0/DNA, Complementary; 0/DNA, Neoplasm; 86090-08-6/Angiostatins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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