摘要
目的 :通过沉默肿瘤坏死因子α诱导的蛋白8(tumor necrosis factor alpha induced protein 8,TNFAIP8,TIPE)的表达,研究其对宫颈癌He La细胞生物学特性的影响,并探讨TNFAIP8在He La细胞增殖、迁移及凋亡中可能的作用机制。方法 :构建特异性针对TNFAIP8基因的慢病毒载体,感染He La细胞后沉默TNFAIP8基因的表达,分别采用实时荧光定量PCR及蛋白质印迹法检测TNFAIP8 m RNA及蛋白在He La细胞中的表达情况。划痕实验及平板克隆形成实验检测沉默TNFAIP8基因表达对He La细胞的迁移和增殖能力的影响。MTT法及锥虫蓝染色计数法检测血清饥饿处理后He La细胞的存活能力,FCM法检测血清饥饿状态下He La细胞的凋亡情况。结果 :实时荧光定量PCR及蛋白质印迹法检测结果表明,TNFAIP8-shR NA转入HeL a细胞后,TNFAIP8 mR NA(P<0.05)及蛋白的表达水平均被明显下调。划痕实验和克隆形成实验结果显示,沉默TNFAIP8基因在HeL a细胞中的表达,明显抑制了HeL a细胞的迁移和增殖能力(P值均<0.05)。血清饥饿处理后的HeL a细胞中,TNFAIP8沉默组细胞的存活能力显著降低,而凋亡率显著增加(P值均<0.05)。结论 :TNFAIP8作为一种致癌基因能促进人宫颈癌He La细胞的迁移和增殖,并抑制He La细胞的凋亡。
Objective： To investigate the effects of tumor necrosis factor alpha induced protein 8 （TNFAIP8） gene silencing on proliferation, migration and apoptosis of cervical carcinoma HeLa cells in vitro, and to explore its possible mechanism. Methods： Lentiviral vector including TNFAIPS-shRNA was constructed then transfected into HeLa cells by virus infection. The expression levels of TNFAIP8 mRNA and protein in HeLa cells were detected by real-time fluorescent quantitative-PCR and Western blotting, respectively. The migration and proliferation abilities of HeLa cells after transfection with TNFAIP8-shRNA were measured by wound healing assay and plate colony formation, respectively. The survival and apoptosis of HeLa cells subjected to serum starvation were tested by MTT method, trypan blue staining and FCM. Results： The results of real-time fluorescent quantitative-PCR and Western blotting showed that the expression levels of TNFAIP8 mRNA and protein in HeLa cells after infection with TNFAIP8-shRNA were down-regulated. The migration and proliferation abilities of HeLa cells with TNFAIP8 gene silencing were significantly inhibited （both P 〈 0.05）. As compared with the negative control group, when subjected to serum starvation, the survival ability of HeLa cells with TNFAIP8 gene silencing was significantly decreased, and the apoptosis rate was significantly increased （both P 〈 0.05）. Conclusion： TNFAIP8, which is an oncogene, can cells, as well as inhibit the apoptosis of HeLa cel promote migration and proliferation of HeLa Is.
出处
《肿瘤》
CAS
CSCD
北大核心
2015年第11期1208-1215,共8页
Tumor
关键词
宫颈肿瘤
肿瘤坏死因子α诱导的蛋白8
RNA
小分子干扰细胞增殖
细胞凋亡
Uterine cervical neoplasms
Tumor necrosis factor alpha induced protein 8RNA, small interfering
Cell proliferation
Apoptosis
