顺铂通过增加HeLa人宫颈癌细胞中Nrf2入核促进细胞自噬和凋亡

季静; 张晔; 陆晓红; 徐锋; 詹惠英

doi:10.13898/j.cnki.issn.1000-2200.2023.03.005

顺铂通过增加HeLa人宫颈癌细胞中Nrf2入核促进细胞自噬和凋亡

Cisplatin increases nuclear translocation of Nrf2 and promotes autophagy and apoptosis in HeLa human cervical cancer cells

摘要

摘要:
目的探讨顺铂对HeLa人宫颈癌细胞中Nrf2蛋白表达及核转位的影响，并观察细胞自噬和凋亡的变化。
方法使用不同浓度的顺铂(0、2.5、5、10 μmol/L)干预HeLa人宫颈癌细胞，MTT法检测细胞活性，Transwell实验观察细胞迁移能力，mCherry-GFP-LC3B融合蛋白腺病毒转染HeLa细胞观察自噬水平的变化，流式细胞术检测细胞凋亡水平，免疫蛋白印迹检测Nrf2在细胞质和细胞核内的表达情况，免疫荧光法观察Nrf2的核转位情况。
结果与对照组相比，2.5、5、10 μmol/L的顺铂均可抑制HeLa细胞的活性及细胞迁移能力(P < 0.05)，且随着药物浓度增高抑制作用增强(P < 0.05)。与对照组相比，2.5、5、10 μmol/L的顺铂均可促进HeLa细胞中LC3B的表达、细胞凋亡及Nrf2的核转位(P < 0.05), 降低细胞质中Nrf2的蛋白表达水平(P < 0.05)，并升高细胞核中Nrf2的蛋白表达水平(P < 0.05)。
结论顺铂通过增加HeLa人宫颈癌细胞中Nrf2入核促进细胞自噬和细胞凋亡，并抑制细胞的活性和迁移能力。

Abstract:
ObjectiveTo investigate the effect of cisplatin on the protein expression and nuclear translocation of Nrf2, and on the changes of autophagy and apoptosis in HeLa human cervical cancer cells.
MethodsAfter the HeLa human cervical cancer cells were treated with different concentrations of cisplatin(0, 2.5, 5 and 10 μmol/L), MTT assay was used to detect the cell activity, Transwell assay was used to observe the cell migration ability, mCherry-GFP-LC3B fusion protein adenovirus transfected HeLa cells were applied to observe the autophagy changes, flow cytometry was employed to detect the apoptosis, Western blotting was used to detect the expression of Nrf2 in the cytoplasm and nucleus, and immunofluorescence staining was used to observe the nuclear translocation of Nrf2.
ResultsCompared with the control group, 2.5, 5 and 10 μmol/L cisplatin could inhibit the activity and migration ability of HeLa cells(P < 0.05), and the inhibitory effect increased with the increase of drug concentration(P < 0.05).Compared with the control group, 2.5, 5 and 10 μmol/L cisplatin could promote the expression of LC3B, apoptosis and nuclear translocation of Nrf2 in HeLa cells(P < 0.05), reduce the protein expression level of Nrf2 in the cytoplasm(P < 0.05), and increase the protein expression level of Nrf2 in the nucleus(P < 0.05).
ConclusionsCisplatin promotes autophagy and apoptosis by increasing nuclear translocation of Nrf2 in HeLa human cervical cancer cells, and inhibits cell viability and migration ability.

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