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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 investigate the expression of Nrf2 protein in cervical squamous cell carcinoma of Uygur and its effect on the biological behavior of cervical cancer cells such as proliferation, migration, invasion and apoptosis. Methods The expression of Nrf2 and Keap1 protein in cervical squamous cell carcinoma, cervical intraepithelial neoplasia (CIN) and normal cervical tissues were detected by immunohistochemistry. At the cellular level, the gene expression of NFE2L2 in SiHa cells was down-regulated by gene silencing, and the corresponding changes of NFE2L2 mRNA and protein levels were detected by qRTPCR and Western blotting. Transwell chamber assay and flow cytometry were used to detect the expression of NFE2L2-SiHa Cell migration, invasion, apoptosis and cell cycle changes. Results Immunohistochemistry showed that the positive expression rate of Nrf2 protein in Uighur cervical squamous cell carcinoma was significantly higher than that in CIN and normal cervical tissues (66.3% vs 27.3%, P <0.001) siRNA downregulated the expression of NFE2L2 gene in SiHa cells, and the expression of NFE2L2 mRNA and protein decreased in SiHa cells (103.00 ± 37.58 / 125.33 ± 34.67 and 82.25 ± 23.56 / 54.75 ± 9.07 ) Compared with the blank control group and the negative control group (P <0.01). The cell cycle arrest in G0 / G1 phase was significantly increased (P <0.01). Conclusion The activation of Keap1-Nrf2 pathway exists in Uighur cervical squamous cell carcinoma. NFE2L2 gene silencing can significantly inhibit the proliferation, migration and invasion of cervical cancer SiHa cells and promote apoptosis.