Adenovirus mediated antisense c-myc gene on the chemotherapy sensitivity of osteosarcoma cells to cisplatin / 中华外科杂志
Zhonghua Wai Ke Za Zhi
; (12): 799-802, 2005.
Article
em Zh
| WPRIM
| ID: wpr-306206
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct the recombinant adenovirus encoding antisense c-myc fragment and to investigate its effect on the chemotherapy sensitivity of osteosarcoma MG-63 cells to cisplatin.</p><p><b>METHODS</b>The recombinant adenovirus (Ad-Asc-myc) encoding antisense c-myc fragment was constructed by cloning c-myc cDNA of about 720 base pairs in a reverse direction into adenovirus vector, then undergoing recombination, amplifying and being complemented in vivo. The osteosarcoma MG-63 cells were transfected by the Ad-Asc-myc in vitro, and Wright staining, Acridine Orange staining, Western Blot, MTT, Flow Cytometry (FCM) were used to study cell morphology, expression of c-myc protein, tumor cell proliferation in vitro, apoptosis and cell cycle change.</p><p><b>RESULTS</b>Ad-Asc-myc encoding antisense c-myc fragment was obtained with the titer of 2 x 10(9) pfu/ml. Ad-Asc-myc down-regulated the expression of c-myc protein after transfected MG-63 cells for 48 h, combined with the treatment of 2.0, 5.0 microg/ml cisplatin for 2 h could inhibit tumor cells proliferation in vitro by 33.4% and 54.2% respectively, which were significantly difference compared with control recombinant adenovirus (Ad-LacZ) groups (P < 0.05). Acridine Orange staining 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 G2/M phase arrested in transfected cells.</p><p><b>CONCLUSION</b>Ad-Asc-myc increases the chemotherapy sensitivity of osteosarcoma MG-63 cells to cisplatin as well as induced apoptosis.</p>
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Assunto principal:
Patologia
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Farmacologia
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Recombinação Genética
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Células Tumorais Cultivadas
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Transfecção
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Osteossarcoma
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Adenovírus Humanos
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DNA Antissenso
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Genes myc
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Proteínas Proto-Oncogênicas c-myc
Limite:
Humans
Idioma:
Zh
Revista:
Zhonghua Wai Ke Za Zhi
Ano de publicação:
2005
Tipo de documento:
Article