| Recombinant antisense C-myc adenovirus increase in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin. | |
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MedLine Citation:
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PMID: 16466985 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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C-myc is an oncogene with the important role of cell proliferation controller. It has been found to be amplified and overexpressed in osteosarcoma. Moreover, it can promote cell transformation and induce metastatic features. Some studies showed that overexpression of c-myc could induce resistance in response to antineoplastic agents. Currently, we constructed the recombinant adenovirus (Ad-Asc-myc) encoding antisense c-myc fragment and investigated its effect on the in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin(CDDP). The osteosarcoma MG-63 cells were transfected by the Ad-Asc-myc in vitro, and Western Blot, MTT assay, RT-PCR, flow cytometry (FCM), and transmission electron microscopy (TEM) were used to study expression of c-myc and caspase-3 protein, tumor cell proliferation in vitro, cell apoptotic morphology and cell cycle change. Ad-Asc-myc encoding antisense c-myc fragment was obtained with the titer of 2.0 x 10(9) pfu/ml. Ad-Asc-myc downregulated the expression of c-myc protein after transfected MG-63 cells for 48 hours, combined with the treatment of 2.0, 5.0 microg/ml cisplatin for 2 hours can inhibited tumor cells proliferation in vitro by 33.4 and 54.2 percent, respectively, which had significant difference compared with control recombinant adenovirus (Ad-LacZ) groups (P < 0.05). RT-PCR revealed that Ad-Asc-myc downregulated expression of bcl-2 and upregulated expression of Bax, and no appreciable changes were observed in the expression of E2F-1. Detection of caspase-3 protein TEM, and FCM analysis showed that Ad-Asc-myc could induce apoptosis of transfected cells, which was enhanced by the treatment of cisplatin. Cell cycle analysis showed that obvious G(2)/M phase arrested in transfected cells. In conclusion, Ad-Asc-myc increased the in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin as well as induced apoptosis. |
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Authors:
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Xian-Kuan Xie; Di-Sheng Yang; Zhao-Ming Ye; Hui-Min Tao |
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Publication Detail:
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Type: In Vitro; Journal Article |
Journal Detail:
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Title: Cancer investigation Volume: 24 ISSN: 0735-7907 ISO Abbreviation: Cancer Invest. Publication Date: 2006 Feb |
Date Detail:
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Created Date: 2006-02-09 Completed Date: 2006-04-13 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8307154 Medline TA: Cancer Invest Country: United States |
Other Details:
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Languages: eng Pagination: 1-8 Citation Subset: IM |
Affiliation:
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Department of Orthopaedics, The Second Hospital Affiliated Zhejiang University, College of Medicine, Hangzhou, People's Republic of China. xiexk2@163.com |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Adenoviridae
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genetics Antineoplastic Agents / pharmacology* Apoptosis / drug effects, physiology Blotting, Western Caspase 3 Caspases / biosynthesis Cell Cycle / drug effects, physiology Cell Line, Tumor Cell Proliferation / drug effects Cisplatin / pharmacology* DNA, Antisense / pharmacology* DNA, Recombinant / pharmacology Flow Cytometry Gene Therapy* Humans Microscopy, Electron, Transmission Osteosarcoma / genetics, therapy*, ultrastructure Proto-Oncogene Proteins c-myc / metabolism* Reverse Transcriptase Polymerase Chain Reaction Transfection |
| Chemical | |
Reg. No./Substance:
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0/Antineoplastic Agents; 0/DNA, Antisense; 0/DNA, Recombinant; 0/MYC protein, human; 0/Proto-Oncogene Proteins c-myc; 15663-27-1/Cisplatin; EC 3.4.22.-/CASP3 protein, human; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases |
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