核因子E2相关因子1过表达抑制宫颈癌细胞增殖及迁移

Overexpression of nuclear factor erythroid 2-related factor 1 inhibitsthe proliferation and migration of cervical cancer cells

目的:初步探究核因子E2相关因子1(nuclear factor erythroid 2-related factor 1,Nrf1)在宫颈癌组织中的表达及其过表达对宫颈癌细胞增殖以及迁移的影响。方法:通过TCGA数据分析Nrf1在人宫颈癌及正常宫颈组织中的表达水平差异;检索OncoLnc网站,比较Nrf1高表达与低表达宫颈癌患者的生存率;采用qRT-PCR检测11对宫颈癌及癌旁组织中Nrf1 mRNA的表达水平。通过慢病毒转染方式构建Nrf1过表达宫颈癌HeLa、SiHa细胞株,以及对应的空载体细胞株;采用qRT-PCR及蛋白免疫印迹实验分别检测细胞株内Nrf1的mRNA和蛋白表达水平;采用EdU细胞增殖实验、细胞克隆形成实验、Transwell细胞迁移实验以及细胞划痕愈合实验检测宫颈癌细胞的增殖和迁移。结果:TCGA数据分析显示,宫颈癌组织中Nrf1 mRNA表达水平明显低于正常宫颈组织(P<0.01);OncoLnc网站分析显示,Nrf1高表达组患者生存率明显高于Nrf1低表达组(P<0.05);qRT-PCR结果显示,Nrf1 mRNA在宫颈癌组织中的表达水平显著低于癌旁组织(P<0.05)。qRT-PCR及蛋白免疫印迹实验结果显示,在宫颈癌HeLa、SiHa细胞株中,Nrf1过表达组Nrf1 mRNA及蛋白水平较空载体对照组明显升高(P<0.05);与空载体对照组相比,Nrf1过表达组EdU标记的细胞占比、克隆细胞数及细胞迁移数均明显降低(P<0.01或P<0.05)。结论:Nrf1在宫颈癌组织中呈低表达;外源性过表达Nrf1可显著抑制宫颈癌细胞的增殖及迁移能力。

Objective:To explore the expression of nuclear factor erythroid 2-related factor 1(Nrf1)in cervical cancer,and the effect of overexpression Nrf1 on proliferation and migration of cervical cancer cells.Methods:TCGA database was applied to analyze the expression levels of Nrf1 in human cervical cancer and normal cervical tissues.OncoLnc website data was used to compare the survival rates of patients with Nrf1 high-and low-expression.The mRNA expression levels of Nrf1 in 11 pairs of cervical cancer and adjacent tissues were detected by qRT-PCR.Lentivirus transfection method was used to construct HeLa and SiHa cell lines with Nrf1 overexpression and corresponding empty vector cell lines.The mRNA and protein expression levels of Nrf1 were detected by qRT-PCR and Western blotting,respectively.EdU cell proliferation assay,cell clone formation assay,Transwell assay and cell wound healing assay were used to evaluate the proliferation and migration of cervical cancer cells.Results:TCGA data analysis showed that the mRNA expression level of Nrf1 in cervical cancer tissues was significantly lower than that in normal cervical tissues(P<0.01).Analysis of OncoLnc website showed that the survival rate of patients in Nrf1 high-expression group was significantly higher than that in Nrf1 low-expression group(P<0.05).The results of qRT-PCR showed that the expression level of Nrf1 mRNA in cervical cancer tissues was significantly lower than that in adjacent tissues(P<0.05).The results of qRT-PCR and Western blotting showed that in cervical cancer HeLa and SiHa cell lines,the mRNA and protein levels of Nrf1 in Nrf1 overexpression group were significantly higher than those in empty vector control group(P<0.05);compared with the empty vector control group,the EdU labelled cell proportion,the number of cloned cells and migration in Nrf1 overexpression group were significantly decreased(P<0.01 or P<0.05).Conclusion:Nrf1 is lowly expressed in cervical cancer tissues.Exogenous overexpression of Nrf1 could significantly inhibit the proliferation and migration of cervical cancer cells.