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

doi:10.16098/j.issn.0529-1356.2019.04.012

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

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

1.武汉科技大学城市学院医学部实验室，武汉 430083; 2.武汉科技大学附属天佑医院妇产科，武汉 430064; 3.湖北省妇幼保健院产科，湖北武汉 430070

收稿日期:2018-06-27 修回日期:2018-07-12 出版日期:2019-08-06 发布日期:2019-08-06
通讯作者: 梅燕 E-mail:my42562@163.com

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

1.Laboratory of Medicine Department, Wuhan University of Science and Technology City College, Wuhan 430083, China; 2.Department of Obstetrics and Gynecology, Tianyou Hospital Affiliated to Wuhan University of Science and Technology, Wuhan 430064, China; 3.Obstetrics Department of Obstetrics, Hubei Maternal and Child Health Hospital, Wuhan 430070, China

Received:2018-06-27 Revised:2018-07-12 Online:2019-08-06 Published:2019-08-06
Contact: MEI Yan E-mail:my42562@163.com

摘要/Abstract

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.

Key words: Nuclear factor erythroid-2-related factor 2, Grainyhead-like 2 gene, Ovarian cancer, Chromatin immunoprecipitation assay, Luciferase reporter gene assay

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.

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.

参考文献

［1］ Gao M, Shih Ie M, Wang TL. The role of forkhead box Q1 transcription factor in ovarian epithelial carcinomas［J］. Int J Mol Sci,2012,13(11):13881-13893.

［2］ Oommen D, Yiannakis D, Jha AN. BRCA1 deficiency increases the sensitivity of ovarian cancer cells to auranofin［J］. Mutat Res, 2016， 784-785(11): 8-15.

［3］ Luo BH, Xiong F, Wang JP, et al. Epidermal growth factor-like domain-containing protein 7 (EGFL7) enhances EGF receptor-AKT signaling, epithelial-mesenchymal transition, and metastasis of gastric cancer cells［J］. PLoS One, 2014， 9(6): e99922.

［4］ Ibanez dCI, Battagli C, Esteller M, et al. Tumor cell-specific BRCA1 and RASSF1A hypermethylation in serum, plasma, and peritoneal fluid from ovarian cancer patients［J］. Cancer Res, 2004， 64(18): 6476-6481.

［5］ Ahmed N, Thompson EW, Quinn MA, Epithelial-mesenchymal interconversions in normal ovarian surface epithelium and ovarian carcinomas: an exception to the norm［J］. J Cell Physiol, 2007，213(3): 581-588.

［6］ Ganan Gomez Ⅰ, Wei Y, Yang H, et al. Oncogenic functions of the transcription factor Nrf2［J］. Free Radic Biol Med, 2013， 65(12): 750-764.

［7］ Tsai JJ, Dudakov JA, Takahashi K, et al. Nrf2 regulates haematopoietic stem cell function［J］. Nat Cell Biol, 2013, 15(3): 309-316.

［8］ Kumar SM， Dey A. Regulation of glutathione in health and disease with special emphasis on chronic alcoholism and hyperglycaemia mediated liver injury: a brief perspective［J］. Springer Sci Rev, 2014，2(1-2): 1-13.

［9］ Kang X, Chen W, Kim RH, et al. Regulation of the hTERT promoter activity by MSH2, the hnRNPs K and D, and GRHL2 in human oral squamous cell carcinoma cells［J］. Oncogene, 2009， 28(4): 565-574.

［10］ Xiang X, Deng Z, Zhuang X, et al. Grhl2 determines the epithelial phenotype of breast cancers and promotes tumor progression［J］. PLoS One, 2013， 8(4): 50781.

［11］ Kajiwara K, Saito A, Ogata SI, et al. Synthetic peptides corresponding to ligand-binding region of death receptors, DR5, Fas, and TNFR, specifically inhibit cell death mediated by the death ligands, respectively［J］. Biochim Biophys Acta, 2004, 1699(1-2):131-137.

［12］ Chen W, Yi JK, Shin KH, et al. Abstract 1598: grainyhead-like 2 (GRHL2) regulates the expression of forkhead box transcription factor M1 (FoxM1) in human oral cancer cells and determines tumorigenicity［J］. Cancer Res, 2014, 74(19 Suppl):1598-1598.

［13］ Xiang J, Fu X, Ran W, et al. Grhl2 reduces invasion and migration through inhibition of TGFβ-induced EMT in gastric cancer［J］. Oncogenesis, 2017, 6(1):e284.

［14］ Kensler TW, Wakabayashi N, Biswal S. Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway.［J］. Annu Rev Pharmacol Toxicol, 2007, 47(1):89-116.

［15］ Arif JM, Sawant SS, Sayed KAE, et al. Antiproliferative potential of sarcophine and its semisynthetic sulfur-containing derivatives against human mammary carcinoma cell lines［J］. J Natl Med, 2006, 61(2):154-158.

［16］ Chen Z, Shentu TP, Wen L, et al.Regulation of SIRT1 by oxidative stress-responsive miRNAs and a systematic approach to identify its role in the endothelium［J］. Antioxid Redox Signal, 2013, 19(13):1522-1535.

［17］ Mooney SM, Talebian Ⅴ, Jolly MK et al. The GRHL2/ZEB feedback loop-a key axis in the regulation of EMT in breast cancer［J］. J Cell Biochem, 2017, 118(9)：112-125.

［18］ Faddaoui A, Sheta R, Bachvarova M, et al.Suppression of the grainyhead transcription factor 2 gene (GRHL2) inhibits the proliferation, migration, invasion and mediates cell cycle arrest of ovarian cancer cells［J］. Cell Cycle, 2017, 16(7): 693-706.

［19］ Kensler TW, Wakabayashi N, Biswal S. Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway［J］. Annu Rev Pharmacol Toxicol, 2007, 47(1):89-116.

［20］ Jaiswal AK. Nrf2 signaling in coordinated activation of antioxidant gene expression ［J］. Free Radic Biol Med, 2004, 36(10):1199-1207.

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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

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

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