| 郑丽端,汪良,童强松,等.Smac基因过表达对宫颈癌HeLa细胞放疗敏感性的影响[J].中华放射医学与防护杂志,2005,25(3):245-247.ZHENG Li-duan,WANG Liang,TONG Qiang-song,et al.Effects of Smac gene over-expression on radiotherapeutic sensitivity of cervical cancer cell line HeLa[J].Chin J Radiol Med Prot,2005,25(3):245-247 |
| Smac基因过表达对宫颈癌HeLa细胞放疗敏感性的影响 |
| Effects of Smac gene over-expression on radiotherapeutic sensitivity of cervical cancer cell line HeLa |
| 投稿时间：2004-05-05 |
| DOI： |
| 中文关键词: 宫颈癌 Smac基因 放疗 凋亡 |
| 英文关键词:Cervical cancer Smac gene Radiotherapy Apoptosis |
| 基金项目: |
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| 中文摘要: |
| 目的 探讨转染Smac基因过表达对宫颈癌细胞放疗敏感性的影响,寻求改善宫颈癌放疗疗效的新途径。方法 将Smac基因转染入宫颈癌HeLa细胞株,G418筛选获得亚克隆细胞,RTPCR、Western blot法检测癌细胞Smac基因表达。X射线照射后,采用MTT比色法检测癌细胞体外生长活性;透射电镜、AnnexinV-FITC和碘化丙啶双染色流式细胞仪法检测癌细胞凋亡及比率;Western blot、比色法检测细胞内Caspase-3蛋白表达和活性水平。结果 RTPCR和Western blot证实所获宫颈癌亚克隆细胞HeLa/Smac的Smac mRNA、蛋白表达水平均显著高于HeLa(P<0.01)。8GyX射线照射后,HeLa/Smac细胞生长活性较HeLa细胞减弱10.19%(P<0.01),透射电镜下可见部分HeLa/Smac细胞发生典型凋亡形态学改变,凋亡率为16.4%,显著高于对照组HeLa细胞(凋亡率为6.2%,P<0.01)。与HeLa细胞比较,HeLa/Smac细胞内Caspase-3表达显著增加(P<0.01),活性水平提高3.42倍(P<0.01)。结论 稳定转染外源性Smac基因使其在宫颈癌细胞株中过表达,能提高癌细胞中Caspase3的表达和活性,增强放疗对癌细胞的诱导凋亡作用,有望成为改善宫颈癌放疗效果的新途径。 |
| 英文摘要: |
| 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. |
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