| ZHENG Li-duan,WANG Liang,TONG Qiang-song.Effects of Smac gene over-expression on radiotherapeutic sensitivity of cervical cancer cell line HeLa[J].Chinese Journal of Radiological Medicine and Protection,2005,25(3):245-247 |
| Effects of Smac gene over-expression on radiotherapeutic sensitivity of cervical cancer cell line HeLa |
| Received:May 05, 2004 |
| DOI： |
| KeyWords:Cervical cancer Smac gene Radiotherapy Apoptosis |
| FundProject: |
|
| Hits: 2620 |
| Download times: 1877 |
| Abstract:: |
| Objective To study the effects of extrinsic Smac gene transfection and its over-expression on radiotherapeutic sensitivity of cervical cancer cells,in order to explore a novel strategy for ameliorating radiotherapy of cervical cancer. Methods After Smac gene was transferred into cells of cervical cancer cell line HeLa,the subclone cells were obtained by persistent G418 selection.Cellular Smac gene expression was determinated by RT-PCR and Western blot.After treatment with X-ray irradiation,cellular growth activity in vitro was investigated by MTT colorimetry.Cellular apoptosis and its rate were determinated by electronmicroscopy,Annexin V-FITC and propidium iodide staining flow cytometry.Cellular Caspase-3 protein expression and its activity were assayed by Western blot and colorimetry. Results RT-PCR and Western blot demonstrated that Smac mRNA and protein levels of HeLa/Smac cells,the selected subclone cells of cervical cancer cell line,were significantly higher than those of HeLa cells (P<0.01).After treated with 8 Gy X-ray irradiation,growth acitivity of HeLa/Smac cells reduced by 10.19%(P<0.01),as compared with that of HeLa cells.Partial HeLa/Smac cancer cells presented characteristic morphological changes of apoptosis under electronmicroscope,with an apoptosis rate of 16.4%,which was significantly higher than that of HeLa cells(6.2%,P<0.01).Compared with HeLa cells,Caspase-3 expression level in HeLa/Smac was improved significantly(P<0.01),while its activity was 3.42 times as much as that of HeLa cells(P<0.01). Conclusion Stable transfection of extrinsic Smac gene and its over-expression in cervical cancer cell line could significantly enhance cellular caspase-3 expression and activity,ameliorate apoptosis-inducing effects of radiation on cancer cells,which would be a novel strategy to improve radiotherapeutic effects for cervical cancer. |
| HTML View Full Text View/Add Comment Download reader |
| Close |
|
|
|
