Adenovirus mediated antisense c-myc gene on the chemotherapy sensitivity of osteosarcoma cells to cisplatin.
- Author:
Di-sheng YANG
1
;
Xian-kuan XIE
;
Zhao-ming YE
;
Hui-min TAO
Author Information
- Publication Type:Journal Article
- MeSH: Adenoviruses, Human; genetics; Antineoplastic Agents; pharmacology; Apoptosis; drug effects; genetics; Cisplatin; pharmacology; DNA, Antisense; genetics; Genes, myc; Genetic Vectors; Humans; Osteosarcoma; genetics; metabolism; pathology; Proto-Oncogene Proteins c-myc; metabolism; Recombination, Genetic; Transfection; Tumor Cells, Cultured
- From: Chinese Journal of Surgery 2005;43(12):799-802
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSThe 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.
RESULTSAd-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.
CONCLUSIONAd-Asc-myc increases the chemotherapy sensitivity of osteosarcoma MG-63 cells to cisplatin as well as induced apoptosis.