Potential role of ZEB1 as a DNA repair regulator in colorectal cancer cells revealed by cancer-associated promoter profiling

Wang,Miao; He,Su-Fei; Liu,Lei-Lei; Sun,Xiao-Xia; Yang,Fan; Ge,Qian; Wong,Wei-Kang; Meng,Jing-Yan

doi:10.3892/or.2017.5888

Potential role of ZEB1 as a DNA repair regulator in colorectal cancer cells revealed by cancer-associated promoter profiling

Authors:
- Miao Wang
- Su-Fei He
- Lei-Lei Liu
- Xiao-Xia Sun
- Fan Yang
- Qian Ge
- Wei-Kang Wong
- Jing-Yan Meng
View Affiliations

Affiliations: College of Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, P.R. China, Collaborative Innovation Center of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, P.R. China
Published online on: August 8, 2017 https://doi.org/10.3892/or.2017.5888
Pages: 1941-1948
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Besides being a key contributor to epithelial‑to‑mesenchymal transition (EMT) activation and stemness maintenance, zinc finger E-box binding homeobox 1 (ZEB1) is also a crucial inducer of chemoresistance and radioresistance. Unlike the clear mechanism that mediates its effect on EMT and dedifferentiation, the mechanism of how ZEB1 promotes chemo- and radio-resistance remains to be elucidated. It has been previously reported that ZEB1 promotes DNA double-strand break clearance by enhancing the deubiquitylating activity of ubiquitin-specific peptidase (USP)7 on checkpoint kinase 1, which is an important step during DNA repair. It was hypothesized that as a transcriptional suppressor, ZEB1 may be involved in an unbalanced DNA damage response (DDR) by affecting other key components. Therefore, in the present study, the target gene occupancy of ZEB1 was mapped in colorectal cancer cells using the ChIP-on-chip method, revealing positive intervals enriched along the three DDR-associated genes: USP17, chromodomain helicase DNA-binding protein 1-like and double homeobox 4. The E-boxes identified in the binding regions and the enhanced mRNA expression of the three genes following the knockdown of ZEB1 supported the identification of these three genes as downstream target genes of ZEB1. Furthermore, ZEB1 knockdown initiated a chemosensitization effect, induced G1/S arrest and increased apoptosis, which functionally validated the three ZEB1 downstream targets. In summary, the present study identified three DDR-associated genes as ZEB1 downstream targets, and demonstrated that their suppression by ZEB1 contributes to ZEB1-mediated chemoresistance.

View Figures

View References

1	Singh A and Settleman J: EMT, cancer stem cells and drug resistance: An emerging axis of evil in the war on cancer. Oncogene. 29:4741–4751. 2010. View Article : Google Scholar : PubMed/NCBI
2	Mitra A, Mishra L and Li S: EMT, CTCs and CSCs in tumor relapse and drug-resistance. Oncotarget. 6:10697–10711. 2015. View Article : Google Scholar : PubMed/NCBI
3	Pattabiraman DR and Weinberg RA: Tackling the cancer stem cells - what challenges do they pose? Nat Rev Drug Discov. 13:497–512. 2014. View Article : Google Scholar : PubMed/NCBI
4	Koren E and Fuchs Y: The bad seed: Cancer stem cells in tumor development and resistance. Drug Resist Updat. 28:1–12. 2016. View Article : Google Scholar : PubMed/NCBI
5	Fischer KR, Durrans A, Lee S, Sheng J, Li F, Wong ST, Choi H, El Rayes T, Ryu S, Troeger J, et al: Epithelial-to-mesenchymal transition is not required for lung metastasis but contributes to chemoresistance. Nature. 527:472–476. 2015. View Article : Google Scholar : PubMed/NCBI
6	Sánchez-Tilló E, Lázaro A, Torrent R, Cuatrecasas M, Vaquero EC, Castells A, Engel P and Postigo A: ZEB1 represses E-cadherin and induces an EMT by recruiting the SWI/SNF chromatin-remodeling protein BRG1. Oncogene. 29:3490–3500. 2010. View Article : Google Scholar : PubMed/NCBI
7	Wellner U, Schubert J, Burk UC, Schmalhofer O, Zhu F, Sonntag A, Waldvogel B, Vannier C, Darling D, zur Hausen A, et al: The EMT-activator ZEB1 promotes tumorigenicity by repressing stemness-inhibiting microRNAs. Nat Cell Biol. 11:1487–1495. 2009. View Article : Google Scholar : PubMed/NCBI
8	Chaffer CL, Marjanovic ND, Lee T, Bell G, Kleer CG, Reinhardt F, D'Alessio AC, Young RA and Weinberg RA: Poised chromatin at the ZEB1 promoter enables breast cancer cell plasticity and enhances tumorigenicity. Cell. 154:61–74. 2013. View Article : Google Scholar : PubMed/NCBI
9	Zhang P, Sun Y and Ma L: ZEB1: At the crossroads of epithelial-mesenchymal transition, metastasis and therapy resistance. Cell Cycle. 14:481–487. 2015. View Article : Google Scholar : PubMed/NCBI
10	Arumugam T, Ramachandran V, Fournier KF, Wang H, Marquis L, Abbruzzese JL, Gallick GE, Logsdon CD, McConkey DJ and Choi W: Epithelial to mesenchymal transition contributes to drug resistance in pancreatic cancer. Cancer Res. 69:5820–5828. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zhang P, Wei Y, Wang L, Debeb BG, Yuan Y, Zhang J, Yuan J, Wang M, Chen D, Sun Y, et al: ATM-mediated stabilization of ZEB1 promotes DNA damage response and radioresistance through CHK1. Nat Cell Biol. 16:864–875. 2014. View Article : Google Scholar : PubMed/NCBI
12	Cortez MA, Valdecanas D, Zhang X, Zhan Y, Bhardwaj V, Calin GA, Komaki R, Giri DK, Quini CC, Wolfe T, et al: Therapeutic delivery of miR-200c enhances radiosensitivity in lung cancer. Mol Ther. 22:1494–1503. 2014. View Article : Google Scholar : PubMed/NCBI
13	Zhang P, Wang L, Rodriguez-Aguayo C, Yuan Y, Debeb BG, Chen D, Sun Y, You MJ, Liu Y, Dean DC, et al: miR-205 acts as a tumour radiosensitizer by targeting ZEB1 and Ubc13. Nat Commun. 5:56712014. View Article : Google Scholar : PubMed/NCBI
14	Srivastava M and Raghavan SC: DNA double-strand break repair inhibitors as cancer therapeutics. Chem Biol. 22:17–29. 2015. View Article : Google Scholar : PubMed/NCBI
15	Tian H, Gao Z, Li H, Zhang B, Wang G, Zhang Q, Pei D and Zheng J: DNA damage response - a double-edged sword in cancer prevention and cancer therapy. Cancer Lett. 358:8–16. 2015. View Article : Google Scholar : PubMed/NCBI
16	Skvortsov S, Debbage P, Lukas P and Skvortsova I: Crosstalk between DNA repair and cancer stem cell (CSC) associated intracellular pathways. Semin Cancer Biol. 31:36–42. 2015. View Article : Google Scholar : PubMed/NCBI
17	Fokas E, Prevo R, Hammond EM, Brunner TB, McKenna WG and Muschel RJ: Targeting ATR in DNA damage response and cancer therapeutics. Cancer Treat Rev. 40:109–117. 2014. View Article : Google Scholar : PubMed/NCBI
18	Benada J and Macurek L: Targeting the checkpoint to kill cancer cells. Biomolecules. 5:1912–1937. 2015. View Article : Google Scholar : PubMed/NCBI
19	Weigelt K, Moehle C, Stempfl T, Weber B and Langmann T: An integrated workflow for analysis of ChIP-chip data. Biotechniques. 45:131–140. 2008. View Article : Google Scholar : PubMed/NCBI
20	Ji X, Li W, Song J, Wei L and Liu XS: CEAS: Cis-regulatory element annotation system. Nucleic Acids Res. 34:(Web Server issue). W551–W554. 2006. View Article : Google Scholar : PubMed/NCBI
21	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−delta delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
22	Sekido R, Murai K, Funahashi J, Kamachi Y, Fujisawa-Sehara A, Nabeshima Y and Kondoh H: The delta-crystallin enhancer-binding protein delta EF1 is a repressor of E2-box-mediated gene activation. Mol Cell Biol. 14:5692–5700. 1994. View Article : Google Scholar : PubMed/NCBI
23	Remacle JE, Kraft H, Lerchner W, Wuytens G, Collart C, Verschueren K, Smith JC and Huylebroeck D: New mode of DNA binding of multi-zinc finger transcription factors: deltaEF1 family members bind with two hands to two target sites. EMBO J. 18:5073–5084. 1999. View Article : Google Scholar : PubMed/NCBI
24	Delgado-Díaz MR, Martín Y, Berg A, Freire R and Smits VA: Dub3 controls DNA damage signalling by direct deubiquitination of H2AX. Mol Oncol. 8:884–893. 2014. View Article : Google Scholar : PubMed/NCBI
25	Ahel D, Horejsí Z, Wiechens N, Polo SE, Garcia-Wilson E, Ahel I, Flynn H, Skehel M, West SC, Jackson SP, et al: Poly(ADP-ribose)-dependent regulation of DNA repair by the chromatin remodeling enzyme ALC1. Science. 325:1240–1243. 2009. View Article : Google Scholar : PubMed/NCBI
26	Furgason JM and Bahassi M: Targeting DNA repair mechanisms in cancer. Pharmacol Ther. 137:298–308. 2013. View Article : Google Scholar : PubMed/NCBI
27	Burrows JF, McGrattan MJ, Rascle A, Humbert M, Baek KH and Johnston JA: DUB-3, a cytokine-inducible deubiquitinating enzyme that blocks proliferation. J Biol Chem. 279:13993–14000. 2004. View Article : Google Scholar : PubMed/NCBI
28	Xu H, Wang Z, Jin S, Hao H, Zheng L, Zhou B, Zhang W, Lv H and Yuan Y: Dux4 induces cell cycle arrest at G1 phase through upregulation of p21 expression. Biochem Biophys Res Commun. 446:235–40. 2014. View Article : Google Scholar : PubMed/NCBI
29	Karimian A, Ahmadi Y and Yousefi B: Multiple functions of p21 in cell cycle, apoptosis and transcriptional regulation after DNA damage. DNA Repair (Amst). 42:63–71. 2016. View Article : Google Scholar : PubMed/NCBI
30	Ciccia A and Elledge SJ: The DNA damage response: Making it safe to play with knives. Mol Cell. 40:179–204. 2010. View Article : Google Scholar : PubMed/NCBI
31	Weber AM and Ryan AJ: ATM and ATR as therapeutic targets in cancer. Pharmacol Ther. 149:124–138. 2015. View Article : Google Scholar : PubMed/NCBI
32	Aparicio T, Baer R and Gautier J: DNA double-strand break repair pathway choice and cancer. DNA Repair (Amst). 19:169–175. 2014. View Article : Google Scholar : PubMed/NCBI
33	Cheng W, Su Y and Xu F: CHD1L: A novel oncogene. Mol Cancer. 12:1702013. View Article : Google Scholar : PubMed/NCBI
34	Jung YS, Qian Y and Chen X: Examination of the expanding pathways for the regulation of p21 expression and activity. Cell Signal. 22:1003–1012. 2010. View Article : Google Scholar : PubMed/NCBI
35	Rohleder F, Huang J, Xue Y, Kuper J, Round A, Seidman M, Wang W and Kisker C: FANCM interacts with PCNA to promote replication traverse of DNA interstrand crosslinks. Nucleic Acids Res. 44:3219–3232. 2016. View Article : Google Scholar : PubMed/NCBI