Keap1在肾细胞癌中的表达及其作用

doi:10.6040/j.issn.1671-7554.0.2016.162

摘要/Abstract

摘要： 目的探讨Kelch样ECH相关蛋白-1(Keap1)在肾癌中的表达及其在肾癌发生发展中的作用。方法采用Q-PCR技术检测32例肾癌患者的肿瘤组织和肾癌旁组织中Keap1、核因子E2相关因子2(NFE2L2)的表达;免疫组织化学染色法检测上述肾癌患者肿瘤组织和肾癌旁组织中Keap1、NFE2L2的表达;CCK-8法检测和克隆形成法检测过表达Keap1后对肾癌细胞系ACHN增殖活性以及克隆形成能力的影响。结果实时荧光定量PCR(Q-PCR)结果显示,75.00%患者Keap1 mRNA的表达癌旁组织高于肿瘤组织(P=0.000),62.50%患者NFE2L2 mRNA的表达肿瘤组织高于癌旁组织(P=0.046)。免疫组化结果显示32例肾癌患者中,Keap1蛋白在癌旁组织(87.50%)的表达高于肿瘤组织(25.00%)(P=0.000),NFE2L2蛋白在肿瘤组织(81.25%)的表达高于癌旁组织(46.88%)(P=0.004)。高表达Keap1后,NFE2L2的表达降低,肾癌细胞系ACHN的增殖能力和克隆形成能力降低。结论 Keap1/NFE2L2通路在肾癌的发生和发展中可能发挥重要作用,Keap1的表达对肾癌可能起到抑制作用。

关键词: 肾细胞癌, 细胞增殖, 核因子E2相关因子2, Kelch样ECH相关蛋白-1

Abstract: Objective To investigate the expression of Keap1 and evaluate its role in renal carcinoma. Methods The total RNA of tumor tissues and adjacent tissues were isolated from 32 cases of renal carcinoma treated during May 2014 to April 2015 by TRIzol. The expressions of Keap1 and NFE2L2 were detected by Q-PCR technology and immunohistochemical staining. The changes of proliferation and colony forming ability of ACHN after overexpressed Keap1 were detected by CCK-8 cell proliferation and clone formation assay. Results Q-PCR results showed that the expression of Keap1 mRNA in adjacent tissues in 75% patients was higher than that in tumor tissues(P=0.000), the expression of NFE2L2 mRNA in tumor tissues in 62% patients was higher than that in adjacent tissues(P=0.046). The immunohistochemical results showed that the expression of Keap1 protein in adjacent tissues(87.50%)was higher than that in tumor tissues(25.00%)(P=0.000), and the expression of NFE2L2 protein in tumor tissues(81.25%)was higher than that in adjacent tissues(46.88%)(P=0.004). Overexpression of Keap1 decreased proliferation and colony forming ability of renal cell carcinoma cell line ACHN. Conclusion The abnormal expression of Keap1 in tumor tissues indicates that the Keap1-NFE2L2 pathway may play an important role in renal carcinoma. Keap1 has a probable inhibiting effect on the progression of renal carcinoma.

Key words: Renal carcinoma, Kelch-like ECH-associated protein1, Proliferation, Nuclear factor E2-related factor 2

中图分类号:

R574

史培堃,曾贝妮,吴伟芳,胡晓燕,马天加,周亚滨. Keap1在肾细胞癌中的表达及其作用[J]. 山东大学学报（医学版）, 2017, 55(3): 94-99.

SHI Peikun, ZENG Beini, WU Weifang, HU Xiaoyan, MA Tianjia, ZHOU Yabin. Expression and role of Keap1 in renal cell carcinoma[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2017, 55(3): 94-99.

参考文献

[1] Brookman-May SD, May M, Shariat SF, et al. Time to recurrence is a significant predictor of cancer-specific survival after recurrence in patients with recurrent renal cell carcinoma-results from a comprehensive multi-centre database(CORONA/SATURN-Project)[J]. BJU Int, 2013, 112(7): 909-916.
[2] Huang Y, Li W, Su ZY, et al. The complexity of the Nrf2 pathway: beyond the antioxidant response[J]. J Nutr Biochem, 2015, 26(12): 1401-1413.
[3] OConnell MA, Hayes JD. The Keap1/Nrf2 pathway in health and disease: from the bench to the clinic[J]. Biochem Soc Trans, 2015, 43(4): 687-689.
[4] Leinonen HM, Kansanen E, Polonen P, et al. Role of the Keap1-Nrf2 pathway in cancer[J]. Adv Cancer Res, 2014, 122: 281-320. doi: 10.1016/B978-0-12-420117-0.00008-6.
[5] Leinonen HM, Kansanen E, Polonen P, et al. Dysregulation of the Keap1-Nrf2 pathway in cancer[J]. Biochem Soc Trans, 2015, 43(4): 645-649.
[6] Kobayashi A, Kang MI, Okawa H, et al. Oxidative stress sensor Keap1 functions as an adaptor for Cul3-based E3 ligase to regulate proteasomal degradation of Nrf2[J]. Mol Cell Biol, 2004, 24(16): 7130-7139.
[7] Guo Y, Yu S, Zhang C, et al. Epigenetic regulation of Keap1-Nrf2 signaling[J]. Free RadicBiol Med, 2015, 88(Pt B): 337-349.
[8] Rushmore TH, Morton MR, Pickett CB. The antioxidant responsive element. Activation by oxidative stress and identification of the DNA consensus sequence required for functional activity[J]. J Biol Chem, 1991, 266(18): 11632-11639.
[9] Tian H, Zhang B, Di J, et al. Keap1: One stone kills three birds Nrf2, IKKβ and Bcl-2/Bcl-xL[J]. Cancer Letters, 2012, 325(1): 26-34.
[10] Zhao CR, Gao ZH, Qu XJ. Nrf2-ARE signaling pathway and natural products for cancer chemoprevention[J]. Cancer Epidemiol, 2010, 34(5): 523-533.
[11] Padmanabhan B, Tong KI, Ohta T, et al. Structural basis for defects of Keap1 activity provoked by its point mutations in lung cancer[J]. Mol Cell, 2006, 21(5): 689-700.
[12] Wang R, An J, Ji F, et al. Hypermethylation of the Keap1 gene in human lung cancer cell lines and lung cancer tissues[J]. Biochem Biophys Res Commun, 2008, 373(1): 151-154.
[13] 曹宝山, 朱翔, 陈森, 等. Keap1在非小细胞肺癌中的表达及与化疗疗效相关性的研究[J]. 中国肺癌杂志, 2012, 15(10): 591-596. CAO Baoshan, ZHU Xiang, CHEN Sen, et al. Keap1 expression for predicting the chemoresistance and prognosis of advanced non-small cell lung cancer[J]. Chinese Journal of Lung Cancer, 2012, 15(10): 591-596.
[14] 张凯茹, 闫惠琴, 王燕, 等. Nrf2及其靶基因在人肺腺癌A549顺铂耐药细胞株中的表达和意义[J]. 中国肺癌杂志, 2009, 12(11): 1150-1154. ZHANG Kairu, YAN Huiqin, WANG Yan, et al. Expression and significance of Nrf2 and its target genes in pulmonary adenocarcinoma A549 cells resistant to cisplatin[J]. Chinese Journal of Lung Cancer, 2009, 12(11): 1150-1154.
[15] Ohta T, Iijima K, Miyamoto M, et al. Loss of Keap1 function activates Nrf2 and provides advantages for lung cancer cell growth[J]. Cancer Res, 2008, 68(5): 1303-1309.
[16] Shim GS, Manandhar S, Shin DH, et al. Acquisition of doxorubicin resistance in ovarian carcinoma cells accompanies activation of the NRF2 pathway[J]. Free RadicBiol Med, 2009, 47(11): 1619-1631.
[17] Huang CF, Zhang L, Ma SR, et al. Clinical significance of Keap1 and Nrf2 in oral squamous cell carcinoma[J]. PLoS One, 2013, 8(12): e83479. doi: 10.1371/journal.pone.0083479. eCollection 2013.

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