关键字:组织因子表达影响阿霉素诱导人神经母细胞瘤凋亡的研究
Regulation of Tissue Factor on Doxorubicininduced Apoptosis in Human Neuriblastoma
FANG Jun1,2*,TANG Hao3,XIA Linghui,ZHOU Muxiang4,LEI Ting3,WEI Wenning,HU Yu1,2,SONG Shangjun1
(1.Department of Hematology,Affiliated Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,430022,China;2.Molecular Targeting Laboratory of Hubei Province;3.Department of Neurosurgery,Affiliated Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology;4.Division of Hematology/Oncology,Department of Pediatrics,Emory University School of Medicine,Atlanta,USA)Abstract: Objective To investigate the regulation of tissue factor(TF) on chemotherapeutic reagentinduced apoptosis in human neuroblastoma.Methods The expression of TF was examined by Western blotting.TFpcDNA 3 expression vector was transfected by using lipofectamine 2000.The cytotoxicity of doxorubicin was determined by WST assay.The activation of Caspase3 and PARP induced by doxorubicin were tested by Western blotting.Apoptotic cells were stained by Hochest 33342 and counted under fluorescence inverted microscope.Results Human neuroblastoma cell line SHEP1 expressed high level of TF while SKNSH was with lowTF level.TFpcDNA 3 plasmid was transfected to SKNSH cells and expression of TF significantly increased.Enforced expression of TF decreased the cytotoxicity of doxorubicin in transfected SKNSH cells.Cleavage of Caspase3 and PARP,which are the main effectors of doxorubicininduced apoptosis,also impaired in transfected SKNSH cells with upregulation of TF.TF expression also decreased the apoptotic cell number in TFenforced expressing SKNSH cells(134.33±21.00) contrasted to those control cells transfected with pcDAN 3(232.33±8.84,P<0.05)while exposing to 1 μg/ml doxorubicin for 24 h.Conclusion These results suggest that aberrant expression of TF could inhibit doxorubicin
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