肿瘤细胞和耐药肿瘤细胞中FoxO3/Keap1/Nrf2通路的表达差异

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (8): 859-865.

肿瘤细胞和耐药肿瘤细胞中FoxO3/Keap1/Nrf2通路的表达差异

叶海主¹，陈亚娟²，赵文英¹，刘晓平²

¹皖南医学院第一附属医院弋矶山医院，肿瘤内科，芜湖 241001，安徽；²皖南医学院药学院，芜湖 241001，安徽

收稿日期:2017-05-17 修回日期:2017-06-11 出版日期:2017-08-26 发布日期:2017-08-18
通讯作者: 刘晓平，男，博士，教授，硕士生导师，研究方向：肿瘤分子药理学。 Tel：13855334156 E-mail：liuxiaoping@wnmc.edu.cn
作者简介:叶海主，男，硕士研究生，研究方向：肿瘤内科。 Tel：17855327732 E-mail：yehaizhu0619@163.com
基金资助:
国家自然科学基金（81272485）

Different expression of FoxO3/Keap1/Nrf2 pathway in tumor cells and drug resistant tumor cells

YE Haizhu ¹, CHEN Yajuan², ZHAO Wenying ¹, LIU Xiaoping ²

¹ Department of Medical Oncology, Wannan Medical College Affiliated Yijishan Hospital, Wuhu 241001, AnHui, China; ²School of Pharmacy, Wannan Medical College, Wuhu 241001, AnHui, China

Received:2017-05-17 Revised:2017-06-11 Online:2017-08-26 Published:2017-08-18

摘要/Abstract

摘要：

目的: 检测FoxO3/Keap1/Nrf2信号通路在肿瘤细胞和耐药肿瘤细胞中的表达差异，探讨FoxO3/Keap1/Nrf2信号在肿瘤细胞耐药中的作用。方法: 构建肺癌A549耐药（A549/DDP）和结肠癌HCT-8耐药（HCT-8/5-FU）两种细胞株，耐药细胞对药物的敏感性用MTT法检测；分别用qRT-PCR、Western blot检测FoxO3/Keap1/Nrf2信号通路成员在非耐药和耐药肿瘤细胞中mRNA和蛋白的表达情况并进行比较。结果: A549/DDP的耐药性明显高于A549细胞（P<0.05)，耐药倍数为5.25倍；HCT-8/5-FU耐药性显著高于HCT-8细胞（P<0.05），耐药倍数为31.67倍；两种耐药细胞A549/DDP和HCT-8/5-FU中的FoxO3、Keap1和Akt基因的mRNA表达水平降低而Nrf2和Nqo1基因的mRNA表达水平升高；A549/DDP和HCT-8/5-FU细胞中的Keap1和FoxO3蛋白表达水平降低而p-Akt、Nrf2、Nqo1的蛋白表达水平升高。结论: FoxO3/Keap1/Nrf2信号通路与肿瘤耐药性有一定相关性，为深入研究该通路并探索降低肿瘤耐药性的治疗策略提供参考。

关键词: FoxO3/Keap1/Nrf2信号通路, A549, HCT-8, 耐药

Abstract:

AIM: To detect the different expression of FoxO3/Keap1/Nrf2 pathway in normal and drug resistant tumor cells so as to investigate the effect of FoxO3/Keap1/Nrf2 signal on drug resistance of tumor cell. METHODS: Cell lines of A549 (resistant to cisplatin) and HCT-8 (resistant to 5-FU) were constructed and verified by the MTT assay. The mRNA and protein expressions of FoxO3/Keap1/Nrf2 pathway members in drug-resistant tumor cells and non- drug-resistant tumor cells were evaluated by QRT-PCR and Western blot. RESULTS:The drug resistance of A549/DDP was significantly stronger than A549 cell (P<0.05), the resistance index was 5.25 times; the drug resistance of HCT-8/5-FU was significantly stronger than HCT-8 cell (P<0.05), the resistance index was 31.67 times; the mRNA expressions of FoxO3, Keap1 and Akt in A549/DDP and HCT-8/5-FU cells were decreased compared with normal control cells, while mRNA expression levels of Nqo1 and Nrf2 were increased; the protein expression of FoxO3 and Keap1 in A549 DDP and HCT-8/5-FU cells were decreased compared with normal control cells, while protein expression levels of p-Akt, Nqo1 and Nrf2 were increased. CONCLUSION: FoxO3/Keap1/Nrf2 signaling pathway is related with tumor resistance, which was referential for further study of this pathway and for exploring therapeutic strategies to reduce drug resistance of cancer.

Key words: FoxO3/Keap1/Nrf2 signaling pathway, A549, HCT-8, drug resistance

中图分类号:

R965.2

叶海主，陈亚娟，赵文英，刘晓平. 肿瘤细胞和耐药肿瘤细胞中FoxO3/Keap1/Nrf2通路的表达差异[J]. 中国临床药理学与治疗学, 2017, 22(8): 859-865.

YE Haizhu, CHEN Yajuan, ZHAO Wenying, LIU Xiaoping. Different expression of FoxO3/Keap1/Nrf2 pathway in tumor cells and drug resistant tumor cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(8): 859-865.

参考文献

［1］Palmeira A, Sousa E, Vascon celos MH, et al. Three decades of P-gp inhibitors: skimming through several generations and scaffolds［J］. Curr Med Chem, 2012, 19(13): 1946-2025.
［2］Xia H, Hui KM. Mechanism of cancer drug resistance and the involvement of noncoding RNAs［J］. Curr Med Chem, 2014, 21(26): 3029-3041.
［3］Hayden A, Douglas J, Sommerlad M, et al. The Nrf2 transcription factor contributes to resistance to cisplatin in bladder cancer［J］. Urol Oncol, 2014, 32(6): 806-814.
［4］Bao LJ, Jaramillo MC, Zhang ZB, et al. Nrf2 induces cisplatin resistance through activation of autophagy in ovarian carcinoma［J］. Int J Clin Exp Pathol, 2014, 7(4):1502-1513.
［5］Wang XJ, Sun Z, Villeneuve NF, et al. Nrf2 enhances resistance of cancer cells to chemotherapeutic drugs, the dark side of Nrf2［J］. Carcinogenesis, 2008, 29(6): 1235-1243.
［6］Guan L, Zhang L, Gong Z, et al. FoxO3 inactivation promotes human cholangiocarcinoma tumorigenesis and chemoresistance through Keap1-Nrf2 signaling［J］. Hepatology, 2016, 63(6): 1914-1927.
［7］王荇，李相成. 胆管癌姑息治疗的研究进展［J］. 临床肝胆病杂志, 2016, 32(5): 1022-1025.
［8］Akhdar H, Loyer P, Rauch C, et al. Involvement of Nrf2 activation in resistance to 5-fluorouracil in human colon cancer HT-29 cells［J］. Eur J Cancer, 2009, 45(12): 2219-2227.
［9］耿苗, 唐修文. Nrf2/ARE信号通路与胃癌耐药关系的研究进展［J］. 中国细胞生物学学报，2012, 34(11): 1129-1133.
［10］Shim GS, Manandhar S, Shin DH, et al. Acquisition of doxorubicin resistance in ovarian carcinoma cells accompanies activation of the NRF2 pathway［J］. Free Radic Biol Med, 2009, 47(11): 1619-1631.
［11］Hu L, Miao W, Loignon M, et al. Putative chemopreventive molecules can increase Nrf2-regulated cell defense in some human cancer cell lines, resulting in resistance to common cytotoxic therapies［J］. Cancer Chemother Pharmacol, 2010, 66(3): 467-474.
［12］Ren D, Villeneuve NF, Jiang T, et al. Brusatol enhances the efficacy of chemotherapy by inhibiting the Nrf2-mediated defense mechanism［J］. Proc Natl Acad Sci USA, 2011, 108(4): 1433-1438.
［13］Coomans de Brachène A, Demoulin JB. FOXO transcription factors in cancer development and therapy［J］. Cell Mol Life Sci, 2016, 73(6): 1159-1172.
［14］Zhang Y, Gan B, Liu D, et al. FoxO family members in cancer［J］. Cancer Biol Ther, 2014, 12(4): 253-259.
［15］Coomans de Brachène A, Demoulin JB. FOXO transcription factors in cancer development and therapy［J］. Cell Mol Life Sci, 2016, 73(6): 1159-1172.
［16］何时, 张声. FoxO3a与肿瘤的发生和发展［J］. 国际病理科学与临床杂志, 2012, 32(6): 536-540.
［17］Ausserlechner MJ, Salvador C, Deutschmann A, et al. Therapy-resistant acute lymphoblastic leukemia (ALL) cells inactivate FOXO3 to escape apoptosis induction by TRAIL and Noxa［J］. Oncotarget, 2013, 4(7): 995-1007.
［18］Shiota M, Yokomizo A, Kashiwagi E, et al. Foxo3a expression and acetylation regulate cancer cell growth and sensitivity to cisplatin［J］. Cancer Sci, 2010, 101(5): 1177-1185.
［19］Sun Y, Zhao S, Tian H, et al. Depletion of PI3K p85α induces cell cycle arrest and apoptosis in colorectal cancer cells［J］. Oncol Rep, 2009, 22(6): 1435-1441.
［20］Wei Z, Liu Y, Wang Y, et al. Downregulation of Foxo3 and TRIM31 by miR-551b in side population promotes cell proliferation, invasion, and drug resistance of ovarian cancer［J］. Med Oncol, 2016, 33(11): 126.

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