核因子E2相关因子2转录调控粒状头样2基因影响上皮性卵巢癌细胞活力

核因子E2相关因子2转录调控粒状头样2基因影响上皮性卵巢癌细胞活力

梅燕张琳杨红管平

解剖学报 ›› 2019, Vol. 50 ›› Issue (4) : 477-482.

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解剖学报 ›› 2019, Vol. 50 ›› Issue (4) : 477-482. DOI: 10.16098/j.issn.0529-1356.2019.04.012

肿瘤生物学

核因子E2相关因子2转录调控粒状头样2基因影响上皮性卵巢癌细胞活力

梅燕^1* 张琳² 杨红³ 管平³

作者信息 +

Regulatory effect of grainyhead-like 2 gene on proliferation of epithelial ovarian cancer cells by nuclear factor erythroid-2-related factor 2 transcription

MEI Yan ^1* ZHANG Lin²YANG Hong³ GUAN Ping³

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文章历史 +

摘要

目的探讨核因子E2相关因子2（Nrf2）及其下游基因粒状头样2（GRHL2）对上皮性卵巢癌是否影响以及增殖细胞在上皮性卵巢癌细胞中Nrf2对GRHL2基因的调控作用。方法采用染色质免疫共沉淀方法检测Nrf2是否与6种候选基因结合。采用Nrf2过表达结合荧光素酶报告基因实验，检测Nrf2对GRHL2基因的转录激活作用。采用Real-time PCR实验，分析卵巢癌患者输卵管上皮组织及不同卵巢癌细胞中Nrf2和GRHL2在mRNA水平的表达。采用Western blotting技术，分析Nrf2过表达和敲降对GRHL2蛋白表达量的影响。采用细胞计数试剂盒-8（CCK-8）实验检测Nrf2过表达和敲降对卵巢癌细胞活力的影响。结果 Nrf2可以与GRHL2启动子结合，对GRHL2基因有转录激活的作用。Nrf2过表达和敲降分别升高和降低GRHL2蛋白质表达量；并且，在卵巢癌患者输卵管上皮组织及不同类型卵巢癌细胞系中Nrf2和GRHL2 mRNA水平均异常升高；Nrf2过表达和敲降分别升高和降低卵巢癌细胞活力，而GRHL2敲降可显著降低Nrf2过表达对卵巢癌细胞活力的增强作用。结论转录因子Nrf2可通过转录激活GRHL2基因，发挥对卵巢癌细胞活力的增强作用。

Abstract

Objective To study the effect of nuclear factor erythroid-2-related factor 2(Nrf2) and the downstream gene grainyhead-like 2(GRHL2) on epithelial ovarian cancer cell lines and the interaction of Nrf2 and GRHL2. Methods We conducted ChIP-PCR assay to test the binding of Nrf2 with six candidate genes including GRHL2. Furthermore, we proved whether Nrf2 could bind with the GRHL2 promoter and transcriptionally activate the GRHL2 gene or not. Moreover, if the overexpression and knockdown of Nrf2 could increase and decrease the GRHL2 protein respectively. Results We discovered that fallopian tube epithelial cells taken from epithelial ovarian cancer patients and ovarian cancer cell lines highly expressed the Nrf2 and GRHL2 at mRNA level. The overexpression and knockdown of Nrf2 could increase and decrease the cells activity, respectively. However, the knockdown of GRHL2 could inhibit the influence of Nrf2 overexpression. Conclusion Nrf2 promotes the activity of epithelial ovarian cancer cells via modulating the GRHL2 gene transcriptionally.

关键词

核因子E2相关因子2 / 粒状头样2基因 / 卵巢癌 / 染色质免疫共沉淀 / 荧光素酶报告基因实验

Key words

Nuclear factor erythroid-2-related factor 2 / Grainyhead-like 2 gene / Ovarian cancer / Chromatin immunoprecipitation assay / Luciferase reporter gene assay

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导出引用

梅燕张琳杨红管平. 核因子E2相关因子2转录调控粒状头样2基因影响上皮性卵巢癌细胞活力[J]. 解剖学报. 2019, 50(4): 477-482 https://doi.org/10.16098/j.issn.0529-1356.2019.04.012

MEI Yan ZHANG Lin YANG Hong GUAN Ping. Regulatory effect of grainyhead-like 2 gene on proliferation of epithelial ovarian cancer cells by nuclear factor erythroid-2-related factor 2 transcription[J]. Acta Anatomica Sinica. 2019, 50(4): 477-482 https://doi.org/10.16098/j.issn.0529-1356.2019.04.012

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