[Effect of EPHA2-siRNA plasmid on biological behavior of human osteosarcoma cells in vitro].
OBJECTIVE
To investigate the role of EPHA2 in regulating apoptosis, proliferation and vasculogenic mimicry of osteosarcoma cells, by gene silencing through RNA interference.


METHODS
EPHA2-siRNA plasmids were achieved by gene cloning. The plasmids were transfected into human osteosarcoma cells (MG63). The expression level of EPHA2 protein was measured by Western blotting. The proliferation, apoptosis and vasculogenic mimicry features of osteosarcoma MG63cells were assessed by light microscopy, MTIP assay, flow cytometry, annexin V-FITC/PI and HE staining, respectively.


RESULTS
The EPHA2-siRNA plasmid was confirmed by DNA sequencing. After treatment with Sequence-specific siRNA targeted EPHA2, the protein level of the transfected group decreased significantly. As compared to non-siRNA transfected cells, the transfected group showed lower proliferation, higher and earlier apoptosis and less osteosarcoma-generated vasculogenic mimicry.


CONCLUSIONS
EPHA2 gene may be involoved in apoptosis and proliferation of osteosarcoma cells, and may be necessary for vasculogenic mimicry. Down-regulation of EPHA2 expression by sequence-specific siRNA may be considered as a new option in the treatment of EPHA2 over-expressing cancer including osteosarcoma in future.
Zhang
Li-zhi
LZ
The Affiliated Tongji Hospital of Tongji University, Shanghai 200065, China.
Cai
Xuan-song
XS
Qian
Zhi-kang
ZK
Mei
Jiong
J
Ma
Xiao-hui
XH
Huang
Huang-yuan
HY
Huang
Wei-da
WD
D017209:Apoptosis; D001859:Bone Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D009389:Neovascularization, Pathologic / Q000473:pathology; D012516:Osteosarcoma / Q000378:metabolism / Q000473:pathology*; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D036104:Receptor, EphA2 / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D034741:RNA, Small Interfering; D036104:Receptor, EphA2