Abstract | BACKGROUND: MATERIALS AND METHODS: The three human cell lines (U87, BT325, SHG44) and the C6 rat glioma cell line were cultured. To study the uptake of oligodeoxynucleotides (ODNs) by glioma cells in vitro, cultured glioma cells readily incorporated caroboxyfluorescein-5-succimidyl ester (FAM) labeled phosphorothioate oligodeoxynucleotides, as demonstrated by immunofluorescence microscopy and flow cytometry. To study the effect of ASODNs treatment on c-Met expression in vitro, Expression of c-Met was assessed by immunofluorescence microscopy and reverse transcriptase polymerase chain reaction (RT-PCR) analysis. For animal studies of ODNs toxicity and uptake, eight rats underwent placement of cisternal catheters, under general anesthesia. Four rats were given 24 mug FAM-labeled ASODNs while the others were given a saline control injection. After a 24 h observation period, rats were sacrificed by barbiturate overdose, and their brains were studied. RESULTS: For all cell lines, fluorescence was seen to increase with increasing ASODNs concentration. Cells treated in similar fashion were also analyzed by flow cytometry to graphically illustrate the differing fluorescence. Multiple glioma cell lines were tested, with similar results. c-Met ASODNs was found to be successfully incorporated from the media into cultured human glioma cells, even at concentrations as low as 2 muM. In addition, maintenance of the pH-dependent green fluorescence color, as seen by immunofluorescence microscopy and by using flow cytometry, indicated that the FAM was not contained within lysosomes. Immunofluorescence microscopy and RT-PCR analysis showed decreases in c-Met expression with oligodeoxynucleotides treatment. Uptake into tumor cells was also demonstrated in vivo, with no detectable toxicity at concentrations exceeding expected therapeutic levels. CONCLUSION:
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Authors | Sheng-Hua Chu, Hong Zhang, Yan-Bin Ma, Dong-Fu Feng, Zhi-An Zhu, Xian-Hou Yuan, Zhi-Qiang Li |
Journal | The Journal of surgical research
(J Surg Res)
Vol. 141
Issue 2
Pg. 284-8
(Aug 2007)
ISSN: 0022-4804 [Print] United States |
PMID | 17561117
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Oligodeoxyribonucleotides, Antisense
- Proto-Oncogene Proteins c-met
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Topics |
- Animals
- Cell Line
- Glioma
(drug therapy)
- Humans
- Male
- Oligodeoxyribonucleotides, Antisense
(pharmacokinetics, therapeutic use, toxicity)
- Proto-Oncogene Proteins c-met
(antagonists & inhibitors, genetics)
- Rats
- Rats, Sprague-Dawley
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