Document Detail

C-Met antisense oligodeoxynucleotide inhibits growth of glioma cells.
MedLine Citation:
PMID:  16720163     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: C-Met, a receptor tyrosine kinase, and its ligand, hepatocyte growth factor, are critical in cellular proliferation, motility, and invasion and are known to be overexpressed in gliomas. The aim of our study was therefore to investigate the effect of transfected caroboxyfluorescein-5-succimidyl ester (FAM)-labeled c-Met antisense oligonucleotide (ASODN) on growth of glioma cells. METHODS: Conjugated FAM-labeled c-Met ASODN was encapsulated by LIPOFECTAMINE PLUS Reagent and then added into the human glioma cell line U251. Cultured cells were divided into 5 groups: control group, 500 nmol/L nonsense oligonucleotide (NSODN) group, 250 nmol/L ASODN group, 500 nmol/L ASODN group, and 750 nmol/L ASODN group. The intracellular distribution of c-Met ASODN was observed with fluorescence microscopy; cell growth was detected by methyl thiazole tetrazolium assay. The apoptosis of U251 cells was also examined with a flow cytometer. Semiquantitative reverse transcriptase polymerase chain reaction and Western blot examinations were carried for expression of c-Met messenger RNA (mRNA) and protein. RESULTS: The blue fluorescence was seen in the cytoplast and nuclei of cells of FAM-labeled c-Met ASODN groups with fluorescence microscopy after the cells were treated with FAM-labeled c-Met ASODN-LIPOFECTAMINE PLUS Reagent complex for 3 hours. Antisense (AS) oligonucleotide caused a statistically significant reduction of cell viability (P < .05), whereas NSODN had no such changes. The cell growth was also significantly inhibited by ASODN (P < .05). After transfection, 250, 500, and 750 nmol/L ASODN induced significant apoptotic response, about 4.67% +/- 2.86%, 8.65% +/- 3.18%, and 12.76% +/- 3.15% for 24 hours (P < .05) and 7.79% +/- 1.92%, 11.43% +/- 1.54%, and 15.78% +/- 1.86% for 48 hours (P < .01), respectively. However, 500 nmol/L NSODN did not induce any significant apoptotic response until 48 hours after transfection (P > .05). A significant loss of c-Met mRNA was presented in ASODN-treated cells, and this was not seen in treatment with NSODN. Protein level was significantly decreased 48 hours after c-Met ASODN transfected. CONCLUSIONS: Antisense oligonucleotide targeting c-Met can be identified as a most potent AS compound, which can inhibit cell growth and induce cell apoptosis. This provides evidence that c-Met plays a role in tumor progression of glioma by acting as an oncogene and suggests that c-Met ASODN may provide a novel approach to therapy for human glioma.
Shenghua Chu; Xianhou Yuan; Zhiqiang Li; Pucha Jiang; Jie Zhang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Surgical neurology     Volume:  65     ISSN:  0090-3019     ISO Abbreviation:  Surg Neurol     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-05-24     Completed Date:  2006-07-11     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  0367070     Medline TA:  Surg Neurol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  533-8; discussion 538     Citation Subset:  IM    
Department of Neurosurgery, Zhongnan Hospital of Wuhan University, Wuhan 430071, China.
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MeSH Terms
Apoptosis / drug effects
Blotting, Western
Cell Line, Tumor
Cell Proliferation
Central Nervous System Neoplasms / pathology,  therapy
DNA, Complementary / genetics
Gene Therapy / methods*
Glioma / pathology*,  prevention & control,  therapy*
Microscopy, Fluorescence
Oligoribonucleotides, Antisense / genetics,  metabolism*,  pharmacology*
Proto-Oncogene Proteins c-met / genetics,  metabolism*,  pharmacology*
RNA, Messenger / genetics
Reverse Transcriptase Polymerase Chain Reaction
Reg. No./Substance:
0/DNA, Complementary; 0/Oligoribonucleotides, Antisense; 0/RNA, Messenger; EC Proteins c-met

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