| 双向调控FAT10表达对人宫颈癌SiHa细胞裸鼠移植瘤放射敏感度的影响 |
| 投稿时间：2016-10-18 修订日期：2016-11-15 点此下载全文 |
| 引用本文：钱林华.双向调控FAT10表达对人宫颈癌SiHa细胞裸鼠移植瘤放射敏感度的影响[J].医学研究杂志,2017,46(7):101-106,61 |
| DOI：
10.11969/j.issn.1673-548X.2017.07.025 |
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| 中文摘要:目的 经过临床病理学标本检测人宫颈癌组织FAT10蛋白异常高表达后，探讨上调和下调FAT10基因表达对宫颈癌细胞株SiHa细胞裸鼠移植瘤放射敏感性的影响。方法 采用RT-PCR法检测30例宫颈癌及癌旁组织中FAT10 mRNA的表达水平，FAT10蛋白在人宫颈癌组织中的相对表达水平明显高于癌旁组织，且差异有统计学意义（P<0.05）。通过电穿孔法分别将FAT10 siRNA和重组表达载体pcDNA3.1-FAT10转染宫颈癌SiHa细胞后，G418筛选得到稳定转染细胞系。转染48h后，荧光定量RT-PCR和Western blot法分别检测细胞中FAT10 mRNA和FAT10蛋白表达水平；CCK-8 检测细胞增殖，克隆形成实验观察上调和下调FAT10表达对宫颈癌细胞株SiHa的放射敏感度的影响；TUNEL法检测细胞凋亡影响；裸鼠移植瘤实验检测上调和下调FAT10表达联合X线照射对宫颈癌细胞的生长抑制作用。结果 FAT10蛋白在人宫颈癌组织中的相对表达水平明显高于癌旁组织，且差异有统计学意义（P<0.05），FAT10下调组宫颈癌SiHa细胞内FAT10基因表达明显降低，FAT10上调组明显升高。FAT10下调组宫颈癌SiHa细胞放射敏感度升高，FAT10上调组宫颈癌SiHa细胞放射敏感度降低（P<0.05）。FAT10下调组宫颈癌SiHa细胞增殖被抑制，细胞凋亡率增加（P<0.05）；FAT10上调组SiHa细胞增殖能力明显增强，细胞凋亡率降低（P<0.05），FAT10下调组裸鼠移植瘤平均体积明显小于对照组（P<0.05）；FAT10上调组的瘤体平均体积大于对照组（P<0.05）。20Gy γ射线照射后FAT10下调组瘤体平均体积较照射前明显减小（P<0.05）；而上调组裸鼠皮下种植瘤平均体积较照射前无明显变化（P>0.05）。结论 下调FAT10表达能抑制人宫颈癌SiHa细胞裸鼠移植瘤的生长，增强瘤体的放射敏感度，上调FAT10表达则使肿瘤辐射抗拒。 |
| 中文关键词:宫颈癌 SiHa细胞 FAT10 移植瘤 裸鼠 |
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| Effect of FAT10 Expression on the Radiation Sensitivity of Human Cervical Carcinoma SiHa Cells in Nude Mice |
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| Abstract:Objective After the detection of FAT10 protein in human cervical cancer tissue behind clinical pathological examination, investigate the effect of up regulation and down regulating the expression of FAT10 gene on the radiation sensitivity of cervical cancer cell line SiHa cells in nude mice. Methods The expression levels of FAT10 mRNA in 30 cases cervical cancer and adjacent tissues were detected by RT-PCR, FAT10 siRNA and recombinant expression vector pcDNA3.1-FAT10 were respectively transfected into cervical cancer SiHa cells by electroporation, and stable transfection cell lines was obtained by G418 screening. After transfection 48h, the expression levels of FAT10 mRNA and protein were respectively detected by fluorescence quantitative RT-PCR and Western blot. Cell proliferation was detected with CCK-8. The effect of FAT10 expression on the radiation sensitivity of cervical cancer cell SiHa line was observed by the clonogenic experiment. The cell apoptosis was detected by TUNEL. The growth of cervical cancer cells was detected by Xenograft tumor assay combined with X-ray irradiation. Results The relative expression level of FAT10 protein in human cervical cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant(P<0.05).The expression of FAT10 gene in cervical cancer SiHa cells was significantly lower in FAT10 down group, and the FAT10 up regulation group was significantly higher. The radiation sensitivity of cervical cancer SiHa cells was increased in the FAT10 down group, and the radiation sensitivity of SiHa cells was decreased in the FAT10 up regulation group (P<0.05). FAT10 down group of cervical cancer SiHa cell proliferation was inhibited, the cell apoptosis rate was increased (P<0.05). The proliferation ability of FAT10 cells was significantly enhanced, and the cell apoptosis rate was decreased in the FAT10 up regulation group (P<0.05). The average volume of transplanted tumor in the FAT10 down group was significantly smaller than that of the control group (P<0.05); the average volume of the tumor in the FAT10 up regulation group was larger than that of the control group (P<0.05). The average volume of the tumor was significantly decreased in the FAT10 down group after irradiation of 20Gy gamma ray(P<0.05), and the average volume of subcutaneous implanted tumor in nude mice was not significantly different (P>0.05). Conclusion Downregulation of FAT10 expression can inhibit the growth of human cervical cancer SiHa cells in nude mice, enhance the radiation sensitivity of tumor cells, up regulation of FAT10 expression will make tumor radiation resistance. |
| keywords:Cervical cancer SiHa cell FAT10 Transplanted tumor Nude mice |
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