Document Detail


Recombinant antisense C-myc adenovirus increase in vitro sensitivity of osteosarcoma MG-63 cells to cisplatin.
MedLine Citation:
PMID:  16466985     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Xian-Kuan Xie; Di-Sheng Yang; Zhao-Ming Ye; Hui-Min Tao
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Publication Detail:
Type:  In Vitro; Journal Article    
Journal Detail:
Title:  Cancer investigation     Volume:  24     ISSN:  0735-7907     ISO Abbreviation:  Cancer Invest.     Publication Date:  2006 Feb 
Date Detail:
Created Date:  2006-02-09     Completed Date:  2006-04-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8307154     Medline TA:  Cancer Invest     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1-8     Citation Subset:  IM    
Affiliation:
Department of Orthopaedics, The Second Hospital Affiliated Zhejiang University, College of Medicine, Hangzhou, People's Republic of China. xiexk2@163.com
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MeSH Terms
Descriptor/Qualifier:
Adenoviridae / 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:
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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