中文摘要：

目的探讨FAT10的碳末端双甘酸缺失突变体对宫颈癌He La细胞增殖和凋亡的影响。方法收集宫颈原位癌组织标本和正常宫颈组织标本各5例,Western blotting法检测组织中FAT10蛋白的表达。采用定点突变技术构建pc DNA3.0-flag-FAT10？GG质粒,分别将野生型FAT10、FAT10△GG及空载体（阴性对照）瞬时转染至He La细胞中,以野生型Hela细胞作为空白对照组,采用Western blotting法检测转染效率,CCK-8法检测细胞增殖情况。转染24h后,各组细胞经顺铂诱导凋亡,采用流式细胞仪检测细胞凋亡率。结果 FAT10蛋白在宫颈癌组织中的表达明显高于正常组织。过表达野生型FAT10对宫颈癌细胞的增殖有明显促进作用,但FAT10发生双甘酸突变之后,该作用受到明显抑制。流式细胞仪检测结果显示：与空白对照组（22.7%±4.2%）和阴性对照组（24.1%±3.8%）相比,过表达野生型FAT10组的细胞凋亡率（10.9%±2.0%）明显降低（P〈0.05）,而FAT10？GG组的细胞凋亡率（25.7%±5.1%）无明显变化（P〉0.05）。结论FAT10可通过其碳末端双甘酸结构促进肿瘤细胞增殖,减少凋亡,从而促进肿瘤的发生发展。

英文摘要：

Objective To investigate the effects of FAT10？GG, a carboxyl-terminal diglycine deficient mutant, on the proliferation and apoptosis of cervical cancer cell line HeLa. Methods Specimens of cervical carcinoma in situ and normal cervix tissue, 5 each, were collected. The expressive levels of FAT10 protein in these specimens were detected by Western blotting. Site-directed mutagenesis was applied to construct the mutant pcDNA3.0-flag-FAT10？GG plasmid. The HeLa cells were then transiently transfected with wild-type FAT10, FAT10？GG and empty vector （used as negative control）, and the wild-type HeLa cells served as blank control. The transfection efficiency of FAT10 or FAT10？GG was detected by Western blotting, and cell proliferation was determined by CCK-8 assay. Cisplatin was used to induce cell apoptosis after cells were transfected for 24h, and the cell apoptotic rates of all groups were determined by flow cytometry. Results Western blotting showed a significantly increased expression of FAT10 protein in cervical carcinoma tissues compared with that in normal cervical tissue. Over-expression of wild FAT10 in HeLa cells obviously promoted cell proliferation, but this promotion was significantly inhibited in cells transfected with its diglycine mutant. Compared with blank control group （22.7%±4.2%） and negative control group （24.1%±3.8%）, the apoptotic rate was significantly reduced in wild FAT10 group （10.9%±2.0%, P〈0.05）, while no obvious changes were found in cells transfected with its diglycine deficient mutant （25.7%±5.1%, P〉0.05）. Conclusion FAT10 can promote cell proliferation and inhibit cell apoptosis through its carboxyl-terminal diglycine motif, and it may play an essential role in carcinogenesis and development of cancer.