摘要
目的探究Nrf2蛋白在维吾尔族宫颈鳞癌组织中的表达以及对宫颈癌细胞增殖、迁移、侵袭和凋亡等生物学行为的影响。方法运用免疫组织化学方法检测宫颈鳞癌组织、宫颈上皮内瘤变(CIN)和正常宫颈组织中Nrf2与Keap1蛋白的表达。在细胞水平,运用基因沉默技术下调SiHa细胞中NFE2L2基因的表达,并通过qRTPCR和蛋白印迹法检测NFE2L2mRNA和蛋白水平相应变化;Transwell小室实验及流式细胞术等技术检测转染NFE2L2-siRNA后SiHa细胞迁移、侵袭、凋亡及细胞周期的改变。结果免疫组化结果显示维吾尔族宫颈鳞癌组织中Nrf2蛋白的阳性表达率(66.3%)明显高于CIN组织(56.5%)和正常宫颈组织(27.3%)(P<0.001);转染NFE2L2-siRNA下调宫颈癌SiHa细胞中NFE2L2基因的表达后,NFE2L2 mRNA水平和蛋白水平表达下降,同时SiHa细胞的迁移的细胞数目(103.00±37.58/125.33±34.67)和侵袭数目(82.25±23.56/54.75±9.07)较空白对照组和阴性对照组明显减少(P<0.01),细胞周期阻滞在G0/G1期,SiHa细胞凋亡率显著增加(P<0.01)。结论维吾尔族宫颈鳞癌组织中存在Keap1-Nrf2通路的激活,NFE2L2基因沉默可明显抑制宫颈癌SiHa细胞的增殖、迁移及侵袭能力,并促进细胞凋亡。
Objective To explore the expression of Nrf2 protein in cervical squamous cell carcinoma and its affection on the invasion,migration,cell cycle and apoptosis of cervical cancer cells.Method Expression of Nrf2 and Keap1protein in carcinoma tissues and CIN and normal cervical tissues were detected by immunohistochemistry.The NFE2L2 gene was down-regulated by gene silencing technology in SiHa cell,and the mRNA and protein expression of NFE2L2 was detected by qRT-PCR and western blot respectively.The cell migration,invasion,apoptosis and cell cycle of SiHa cell which was transfected with NFE2L2-siRNA were analyzed by transwell and flow cytometry.Results In Uighur patients,the results of immunohistochemistry showed that the positive rate of Nrf2（66.3%）in cervical squamous carcinoma tissue was obviously higher than in CIN（56.5%）and normal cervical tissue（27.3%）（P〈0.001）.When the expression of NFE2L2 was down-regulated in SiHa cell,the mRNA and protein expression of NFE2L2 were declined,and the migrated cells（103.00±37.58/125.33±34.67）and invasion cells（82.25±23.56/54.75±9.07）were reduced compared to the blank control and negative control.The cell cycle of SiHa were arrested at G0/G1 phase,and the apoptosis rate of SiHa cell was increased significantly（P〈0.01）.Conclusion Keap1-Nrf2 pathway was activated in cervical squamous cell carcinoma of Uighur patient.The gene silence of NFE2L2 could obviously inhibit the cell proliferation,migration and invasion ability,and promote cell apoptosis of cervical cancer cell SiHa.
出处
《新疆医科大学学报》
CAS
2016年第4期411-417,共7页
Journal of Xinjiang Medical University
基金
国家自然科学基金(81360332)
