NRF2/PI3K通路介导肝癌细胞SMMC-7721出现索拉非尼耐药的机制研究

肝脏 ›› 2020, Vol. 25 ›› Issue (9): 930-932.

NRF2/PI3K通路介导肝癌细胞SMMC-7721出现索拉非尼耐药的机制研究

段俊伟, 倪文

431600 湖北汉川市人民医院肿瘤科(段俊伟),急诊科(倪文)

收稿日期:2020-04-08 出版日期:2020-09-30 发布日期:2020-10-22
通讯作者: 倪文
基金资助:
湖北省自然科学基金(2016CBD0218)

A mechanism study on Sorafenib resistance mediated by NRF2/PI3K pathway with a human hepatoma cell line

DUAN Jun-wei¹, NI Wen²

1. Department of Oncology, Hanchuan People's Hospital, Hanchuan, Hubei 431600, China;
2. Emergency Department of Hanchuan People's Hospital, Hanchuan, Hubei Province, 431600

Received:2020-04-08 Online:2020-09-30 Published:2020-10-22
Contact: NI Wen

摘要/Abstract

摘要： 目的探讨NRF2PI3K通路介导肝癌细胞SMMC-7721出现索拉非尼耐药的机制研究。方法建立肝癌细胞系SMMC-7721耐药细胞株SMMC-7721-SR,构建NRF2-siRNA转染肝癌细胞,并给与10 μmol/L索拉非尼处理48 h。采用 CCK-8 法检测细胞活力,采用流式细胞仪检测细胞凋亡, Western blot 检测 NRF2、Keap1、ARE、p-PI3K和p-Akt蛋白表达水平。Real-Time PCR检测NRF2、Keap1、ARE、p-PI3K和p-Akt基因表达水平。结果索拉非尼明显抑制肝癌细胞SMMC-7721的存活率,促进肝癌细胞SMMC-7721 凋亡发生(P<0.05)。索拉非尼对SMMC-7721细胞促凋亡作用明显强于SMMC-7721-SR细胞 (P<0.05)。SMMC-7721-SR细胞中NRF2 蛋白水平及mRNA表达明显高于SMMC-7721细胞,Keap1、ARE、PI3K和Akt蛋白水平及mRNA表达明显增加(P<0.05)。索拉非尼明显上调SMMC-7721细胞中NRF2 蛋白水平及mRNA表达(P<0.05)。抑制NRF2 mRNA表达并给予索拉非尼处理后,SMMC-7721-SR细胞存活率明显降低,凋亡率显著升高,Keap1、ARE、p-PI3K和p-Akt 蛋白水平及mRNA表达明显降低(P<0.05)。结论索拉非尼激活 NRF2后诱导肝癌细胞SMMC-7721出现耐药;抑制 NRF2 可逆转肝癌细胞SMMC-7721对索拉非尼的耐药情况。

关键词: NRF2/PI3K, 肝癌, SMMC-7721, 索拉非尼, 耐药

Abstract: Objective To investigate the involvement of NRF2/PI3K signaling pathway in sorafenib resistance with a liver cancer cell line SMMC-7721. Methods A SMMC-7721 drug-resistant cell line SMMC-7721-SR was first established by screening under ascendant concentrations of sorafinib. The cells were then transfected with NRF2 siRNA and treated with 10 μmol/L sorafenib for 48 h. Cells viability was detected by CCK-8 method. Apoptosis was detected by flow cytometry. The protein levels of NRF2, Keap1, ARE, p-PI3K and p-Akt were detected by Western blot. The mRNA levels of NRF2, Keap1, ARE, PI3K and Akt gene expression were detected by Real-Time quantitative PCR. Results Sorafenib significantly reduced the survival rate and promoted the apoptosis of SMMC-7721 cells (P<0.05). The promoting effect of Sorafenib on the cells apoptosis was more significant in SMMC-7721 cells when compared to the SMMC-77211-SR cells (P<0.05). The expression of NRF2 mRNA and protein in SMMC-7721-SR cells was significantly higher than that in SMMC-7721 cells, and the levels of Keap1, ARE, p-PI3K and p-Akt protein were significantly higher in SMMC-7721-SR cells (P<0.05). Sorafenib significantly upregulated NRF2 protein and mRNA expression in SMMC-7721 cells (P<0.05). After inhibiting the expression of NRF2 mRNA by specific siRNA, the survival rate of SMMC-7721-SR cells was significantly decreased, and the apoptosis rate was significantly increased, accompanied by decreases in the levels of Keap1, ARE, p-PI3K and p-Akt protein and mRNA expression in the cells after sorafenib treatment (P<0.05). Conclusion Sorafenib induces drug resistance in SMMC-7721 cells via activating NRF2. Inhibition of NRF2 may reverse the sorafenib resistance in SMMC-7721 cells.

Key words: NRF2/PI3K, Liver cancer, SMMC-7721, Sorafenib, Drug resistance

段俊伟, 倪文. NRF2/PI3K通路介导肝癌细胞SMMC-7721出现索拉非尼耐药的机制研究[J]. 肝脏, 2020, 25(9): 930-932.

DUAN Jun-wei, NI Wen. A mechanism study on Sorafenib resistance mediated by NRF2/PI3K pathway with a human hepatoma cell line[J]. Chinese Hepatolgy, 2020, 25(9): 930-932.

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