|Expression of angiostatin cDNA in human gallbladder carcinoma cell line GBC-SD and its effect on endothelial proliferation and growth.|
|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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|Type: Journal Article|
|Title: World journal of gastroenterology : WJG Volume: 12 ISSN: 1007-9327 ISO Abbreviation: World J. Gastroenterol. Publication Date: 2006 May|
|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|
|Languages: eng Pagination: 2762-6 Citation Subset: IM|
|APA/MLA Format Download EndNote Download BibTex|
Carcinoma / chemistry, genetics*, pathology
Cell Line, Tumor
Cell Proliferation / drug effects*
Cell Survival / drug effects
DNA, Complementary / analysis*, genetics
DNA, Neoplasm / analysis*, genetics
Endothelium, Vascular / cytology*, growth & development
Gallbladder Neoplasms / chemistry, genetics*, pathology
Gene Expression Regulation, Neoplastic
Genetic Vectors / genetics
Reverse Transcriptase Polymerase Chain Reaction
|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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