过表达免疫负调控分子肿瘤坏死因子诱导蛋白8样蛋白2对人甲状腺癌细胞FTC-133增殖、凋亡和侵袭的影响

Effect of tumor necrosis factor-α induced protein 8 like 2 over-expression on cell proliferation,apoptosis and invasion in human thyroid carcinoma FTC-133 cells

目的 检测肿瘤坏死因子诱导蛋白8样蛋白2（TIPE2）过表达对甲状腺癌细胞FTC-133增殖、凋亡以及侵袭能力的影响.方法 采用脂质体进行瞬时转染和G418筛选法构建TIPE2基因稳定转染甲状腺癌细胞FTC-133;细胞计数试剂盒（CCK-8）法检测24、48、72 h的细胞增殖,绘制细胞生长曲线,并计算细胞增殖抑制率;膜联蛋白V（Annexin V）/碘化丙锭（PI）法检测细胞凋亡,Transwell法检测细胞的侵袭能力.结果 筛选获得稳定高表达TIPE2的克隆细胞;Westernblot结果显示稳定转染TIPE2的FTC-133可明显上调TIPE2蛋白的表达水平,上调倍数达3.33倍;TIPE2过表达后细胞生长速度较对照组细胞明显减慢,24、48、72 h的细胞增殖抑制率分别为（21.60±5.66）％、（31.30±3.97）％、（40.80士7.37）％;TIPE2过表达后细胞凋亡明显增加,细胞总凋亡率为（13.06±1.89）％,而对照组细胞的总凋亡率为（5.73±0.43）％,两者差异有统计学意义（f=8.671,P ＜0.01）;Transwell实验结果显示表达TIPE2的甲状腺癌细胞穿膜细胞数为（51.3 ±16.7）个,而对照组的穿膜细胞数为（91.7±22.7）个,差异有统计学意义（t=7.916,P＜0.01）.结论 TIPE2能明显抑制甲状腺癌细胞的生长、促进其凋亡,同时抑制肿瘤细胞的侵袭能力.

Objective To investigate the effect of tumor necrosis factor-α induced protein 8 like 2 （TIPE2） over-expression on cell proliferation,apoptosis and invasion in human thyroid carcinoma FTC-133 cells.Methods Stable transfected TIPE2 in FTC-133 cells was constructed using Lipofectamine 2000 transfection and G418 selection.Cell proliferation at 24,48 and 72 h was examined by cell counting kit-8 （CCK-8）,cell growth curve and cell inhibitory rate were evaluated,cell apoptosis was examined by annexin V/propidium iodide （PI）,and the cell invasion was evaluated using matrigel coated transwell assay.Results Western blotting analysis confirmed that TIPE2-stable transfected cells expressed high protein levels of TIPE2,which was 3.33 times higher than mock control cells.TIPE2 over expression inhibited cells growth with inhibitory rate being （21.60 ±5.66） %,（31.30 ± 3.97）%、（40.80 ± 7.37） % at 24,48,72 h,respectively compared with mock control cells.TIPE2 over expression could promote cell apoptosis with apoptosis rate being （13.06 ± 1.89） %,which was significantly higher than mock control cells [（5.73 ± 0.43） %,t =8.671,P 〈 0.01].The number of cells invading through matrigel was higher in the TIPE2 over-expressed cells （51.3 ± 16.7） than that in mock-transfected cells （91.7 ±22.7,t =7.916,P 〈 0.01）.Conclusion TIPE2 inhibited thyroid tumor growth,promoted tumor apoptosis and inhibited tumor cells invasion.