Checkpoint kinase‑1 inhibition and etoposide exhibit a strong synergistic anticancer effect on chronic myeloid leukemia cell line K562 by impairing homologous recombination DNA damage repair

Fan,Zhuoyi; Luo,Huacheng; Zhou,Jie; Wang,Fangce; Zhang,Wenjun; Wang,Jian; Li,Shuo; Lai,Qian; Xu,Yueshuang; Wang,Guangming; Liang,Aibin; Xu,Jun

doi:10.3892/or.2020.7757

Checkpoint kinase‑1 inhibition and etoposide exhibit a strong synergistic anticancer effect on chronic myeloid leukemia cell line K562 by impairing homologous recombination DNA damage repair

Authors:
- Zhuoyi Fan
- Huacheng Luo
- Jie Zhou
- Fangce Wang
- Wenjun Zhang
- Jian Wang
- Shuo Li
- Qian Lai
- Yueshuang Xu
- Guangming Wang
- Aibin Liang
- Jun Xu
View Affiliations

Affiliations: Department of Hematology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, P.R. China, Division of Pediatric Hematology/Oncology, Department of Pediatrics, Pennsylvania State University College of Medicine, Hershey, PA 17033, USA, East Hospital, Tongji University School of Medicine, Shanghai 200120, P.R. China, Jake Gittlen Laboratories for Cancer Research, Pennsylvania State University College of Medicine, Hershey, PA 17033, USA, Department of Hematology, The First Affiliated Hospital of Xiamen University and Institute of Hematology, School of Medicine, Xiamen University, Xiamen, Fujian 361003, P.R. China, Department of Pathology, School of Medicine, Southeast University, Nanjing, Jiangsu 210009, P.R. China
Published online on: September 7, 2020 https://doi.org/10.3892/or.2020.7757
Pages: 2152-2164
Copyright: © Fan 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

Leukemia, a malignant hematological disease, has poor therapeutic outcomes due to chemotherapeutic resistance. Increasing evidence has confirmed that the elevated capacity for DNA damage repair in cancer cells is a major mechanism of acquired chemotherapeutic resistance. Thus, combining chemotherapy with inhibitors of DNA damage repair pathways is potentially an ideal strategy for treating leukemia. Checkpoint kinase 1 (CHK1) is an important component of the DNA damage response (DDR) and is involved in the G2/M DNA damage checkpoint. In the present study, we demonstrated that shRNA‑mediated CHK1 silencing suppressed cell proliferation and enhanced the cytotoxic effects of etoposide (VP16) in the chronic myeloid leukemia (CML) cell line K562 through the results of CCK‑8, and comet assay. The results demonstrated that shRNA‑induced CHK1 silencing can override G2/M arrest and impair homologous recombination (HR) repair by reducing breast cancer susceptibility gene 1 (BRCA1) expression. Cells had no time, and thus limited ability, to repair the damage and were thus more sensitive to chemotherapy after CHK1 downregulation. Second, we tested the therapeutic effect of VP16 combined with CCT245737, an orally bioavailable CHK1 inhibitor, and observed strong synergistic anticancer effects in K562 cells. Moreover, we discovered that CCT245737 significantly prevented the G2/M arrest caused by acute exposure to VP16. Interestingly, CCT245737 inhibited both BRCA1 and Rad51, the most important component of the HR repair pathway. In conclusion, these results revealed that CHK1 is potentially an ideal therapeutic target for the treatment of CML and that CCT245737 should be considered a candidate drug.

View Figures

View References

1	Bhamidipati PK, Kantarjian H, Cortes J, Cornelison AM and Jabbour E: Management of imatinib-resistant patients with chronic myeloid leukemia. Ther Adv Hematol. 4:103–117. 2013. View Article : Google Scholar : PubMed/NCBI
2	Kaleem B, Shahab S, Ahmed N and Shamsi TS: Chronic myeloid leukemia-prognostic value of mutations. Asian Pac J Cancer Prev. 16:7415–7423. 2015. View Article : Google Scholar : PubMed/NCBI
3	Valent P, Hadzijusufovic E, Schernthaner GH, Wolf D, Rea D and le Coutre P: Vascular safety issues in CML patients treated with BCR/ABL1 kinase inhibitors. Blood. 125:901–906. 2015. View Article : Google Scholar : PubMed/NCBI
4	Experts in Chronic Myeloid Leukemia, . The price of drugs for chronic myeloid leukemia (CML) is a reflection of the unsustainable prices of cancer drugs: From the perspective of a large group of CML experts. Blood. 121:4439–4442. 2013. View Article : Google Scholar : PubMed/NCBI
5	Dillon MT, Good JS and Harrington KJ: Selective targeting of the G₂/M cell cycle checkpoint to improve the therapeutic index of radiotherapy. Clin Oncol (R Coll Radiol). 26:257–265. 2014. View Article : Google Scholar : PubMed/NCBI
6	Del Nagro CJ, Choi J, Xiao Y, Rangell L, Mohan S, Pandita A, Zha J, Jackson PK and O'Brien T: Chk1 inhibition in p53-deficient cell lines drives rapid chromosome fragmentation followed by caspase-independent cell death. Cell Cycle. 13:303–314. 2014. View Article : Google Scholar : PubMed/NCBI
7	Cimprich KA and Cortez D: ATR: An essential regulator of genome integrity. Nat Rev Mol Cell Biol. 9:616–627. 2008. View Article : Google Scholar : PubMed/NCBI
8	Yazinski SA and Zou L: Functions, regulation, and therapeutic implications of the ATR checkpoint pathway. Annu Rev Genet. 50:155–173. 2016. View Article : Google Scholar : PubMed/NCBI
9	Jazayeri A, Falck J, Lukas C, Bartek J, Smith GC, Lukas J and Jackson SP: ATM- and cell cycledependent regulation of ATR in response to DNA double-strand breaks. Nat Cell Biol. 8:37–45. 2006. View Article : Google Scholar : PubMed/NCBI
10	Sarmento LM, Póvoa V, Nascimento R, Real G, Antunes I, Martins LR, Moita C, Alves PM, Abecasis M, Moita LF, et al: CHK1 overexpression in T-cell acute lymphoblastic leukemia is essential for proliferation and survival by preventing excessive replication stress. Oncogene. 34:2978–2990. 2015. View Article : Google Scholar : PubMed/NCBI
11	Walworth N, Davey S and Beach D: Fission yeast chk1 protein kinase links the rad checkpoint pathway to cdc2. Nature. 363:368–371. 1993. View Article : Google Scholar : PubMed/NCBI
12	Bartek J and Lukas J: Chk1 and Chk2 kinases in checkpoint control and cancer. Cancer Cell. 3:421–429. 2003. View Article : Google Scholar : PubMed/NCBI
13	Zhang Y and Hunter T: Roles of Chk1 in cell biology and cancer therapy. Int J Cancer. 134:1013–1023. 2014. View Article : Google Scholar : PubMed/NCBI
14	Patil M, Pabla N and Dong Z: Checkpoint kinase 1 in DNA damage response and cell cycle regulation. Cell Mol Life Sci. 70:4009–4021. 2013. View Article : Google Scholar : PubMed/NCBI
15	Maya-Mendoza A, Petermann E, Gillespie DA, Caldecott KW and Jackson DA: Chk1 regulates the density of active replication origins during the vertebrate S phase. EMBO J. 26:2719–2731. 2007. View Article : Google Scholar : PubMed/NCBI
16	Tang J, Erikson RL and Liu X: Checkpoint kinase 1 (Chk1) is required for mitotic progression through negative regulation of polo-like kinase 1 (Plk1). Proc Natl Acad Sci USA. 103:11964–11969. 2006. View Article : Google Scholar : PubMed/NCBI
17	Sorensen CS, Hansen LT, Dziegielewski J, Syljuåsen RG, Lundin C, Bartek J and Helleday T: The cell-cycle checkpoint kinase Chk1 is required for mammalian homologous recombination repair. Nat Cell Biol. 7:195–201. 2005. View Article : Google Scholar : PubMed/NCBI
18	Wang X, Kennedy RD, Ray K, Stuckert P, Ellenberger T and D'Andrea AD: Chk1-mediated phosphorylation of FANCE is required for the Fanconi anemia/BRCA pathway. Mol Cell Biol. 27:3098–3108. 2007. View Article : Google Scholar : PubMed/NCBI
19	Walton MI, Eve PD, Hayes A, Valenti MR, De Haven Brandon AK, Box G, Hallsworth A, Smith EL, Boxall KJ, Lainchbury M, et al: CCT244747 is a novel potent and selective CHK1 inhibitor with oral efficacy alone and in combination with genotoxic anticancer drugs. Clin Cancer Res. 18:5650–5661. 2012. View Article : Google Scholar : PubMed/NCBI
20	Calvo E, Braiteh F, Von Hoff D, McWilliams R, Becerra C, Galsky MD, Jameson G, Lin J, McKane S, Wickremsinhe ER, et al: Phase I study of CHK1 inhibitor LY2603618 in combination with gemcitabine in patients with solid tumors. Oncology. 91:251–260. 2016. View Article : Google Scholar : PubMed/NCBI
21	Laquente B, Lopez-Martin J, Richards D, Illerhaus G, Chang DZ, Kim G, Stella P, Richel D, Szcylik C, Cascinu S, et al: A phase II study to evaluate LY2603618 in combination with gemcitabine in pancreatic cancer patients. BMC Cancer. 17:1372017. View Article : Google Scholar : PubMed/NCBI
22	Yin Y, Shen Q, Zhang P, Tao R, Chang W, Li R, Xie G, Liu W, Zhang L, Kapoor P, et al: Chk1 inhibition potentiates the therapeutic efficacy of PARP inhibitor BMN673 in gastric cancer. Am J Cancer Res. 7:473–483. 2017.PubMed/NCBI
23	Italiano A, Infante JR, Shapiro GI, Moore KN, LoRusso PM, Hamilton E, Cousin S, Toulmonde M, Postel-Vinay S, Tolaney S, et al: Phase I study of the checkpoint kinase 1 inhibitor GDC-0575 in combination with gemcitabine in patients with refractory solid tumors. Ann Oncol. 29:1304–1311. 2018. View Article : Google Scholar : PubMed/NCBI
24	Restelli V, Lupi M, Vagni M, Chila R, Bertoni F, Damia G and Carrassa L: Combining ibrutinib with Chk1 inhibitors synergistically targets mantle cell lymphoma cell lines. Target Oncol. 13:235–245. 2018. View Article : Google Scholar : PubMed/NCBI
25	Booth L, Roberts J, Poklepovic A and Dent P: The CHK1 inhibitor SRA737 synergizes with PARP1 inhibitors to kill carcinoma cells. Cancer Biol Ther. 19:786–796. 2018. View Article : Google Scholar : PubMed/NCBI
26	Walton MI, Eve PD, Hayes A, Henley AT, Valenti MR, De Haven Brandon AK, Box G, Boxall KJ, Tall M, Swales K, et al: The clinical development candidate CCT245737 is an orally active CHK1 inhibitor with preclinical activity in RAS mutant NSCLC and Emicro-MYC driven B-cell lymphoma. Oncotarget. 7:2329–2342. 2016. View Article : Google Scholar : PubMed/NCBI
27	Osborne JD, Matthews TP, McHardy T, Proisy N, Cheung KM, Lainchbury M, Brown N, Walton MI, Eve PD, Boxall KJ, et al: Multiparameter lead optimization to give an oral checkpoint kinase 1 (CHK1) inhibitor clinical candidate: (R)-5-((4-((Morpholin-2-ylmethyl)amino)-5-(trifluoromethyl)pyridin-2-yl)amino)pyr azine-2-carbonitrile (CCT245737). J Med Chem. 59:5221–5237. 2016. View Article : Google Scholar : PubMed/NCBI
28	Yang M, Tian X, Fan Z, Yu W, Li Z, Zhou J, Zhang W and Liang A: Targeting RAD51 enhances chemosensitivity of adult T-cell leukemia-lymphoma cells by reducing DNA double-strand break repair. Oncol Rep. 42:2426–2434. 2019.PubMed/NCBI
29	Li L, Ye S, Yang M, Yu W, Fan Z, Zhang H, Hu J, Liang A and Zhang W: SIRT1 downregulation enhances chemosensitivity and survival of adult T-cell leukemia-lymphoma cells by reducing DNA double-strand repair. Oncol Rep. 34:2935–2942. 2015. View Article : Google Scholar : PubMed/NCBI
30	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
31	Chou TC: Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 58:621–681. 2006. View Article : Google Scholar : PubMed/NCBI
32	Tirrò E, Massimino M, Romano C, Pennisi MS, Stella S, Vitale SR, Fidilio A, Manzella L, Parrinello NL, Stagno F, et al: Chk1 inhibition restores inotuzumab ozogamicin citotoxicity in CD22-positive cells expressing mutant p53. Front Oncol. 9:572019. View Article : Google Scholar : PubMed/NCBI
33	Parsels LA, Qian Y, Tanska DM, Gross M, Zhao L, Hassan MC, Arumugarajah S, Parsels JD, Hylander-Gans L, Simeone DM, et al: Assessment of chk1 phosphorylation as a pharmacodynamic biomarker of chk1 inhibition. Clin Cancer Res. 17:3706–3715. 2011. View Article : Google Scholar : PubMed/NCBI
34	Morimoto S, Tsuda M, Bunch H, Sasanuma H, Austin C and Takeda S: Type II DNA topoisomerases cause spontaneous double-strand breaks in genomic DNA. Genes (Basel). 10:8682019. View Article : Google Scholar
35	Schonn I, Hennesen J and Dartsch DC: Cellular responses to etoposide: Cell death despite cell cycle arrest and repair of DNA damage. Apoptosis. 15:162–172. 2010. View Article : Google Scholar : PubMed/NCBI
36	Shibata A and Jeggo PA: DNA double-strand break repair in a cellular context. Clin Oncol (R Coll Radiol). 26:243–249. 2014. View Article : Google Scholar : PubMed/NCBI
37	Chang HHY, Pannunzio NR, Adachi N and Lieber MR: Non-homologous DNA end joining and alternative pathways to double-strand break repair. Nat Rev Mol Cell Biol. 18:495–506. 2017. View Article : Google Scholar : PubMed/NCBI
38	Savage KI, Gorski JJ, Barros EM, Irwin GW, Manti L, Powell AJ, Pellagatti A, Lukashchuk N, McCance DJ, McCluggage WG, et al: Identification of a BRCA1-mRNA splicing complex required for efficient DNA repair and maintenance of genomic stability. Mol Cell. 54:445–459. 2014. View Article : Google Scholar : PubMed/NCBI
39	Roy R, Chun J and Powell SN: BRCA1 and BRCA2: Different roles in a common pathway of genome protection. Nat Rev Cancer. 12:68–78. 2011. View Article : Google Scholar : PubMed/NCBI
40	Moynahan ME, Chiu JW, Koller BH and Jasin M: Brca1 controls homology-directed DNA repair. Mol Cell. 4:511–518. 1999. View Article : Google Scholar : PubMed/NCBI
41	Caestecker KW and Van de Walle GR: The role of BRCA1 in DNA double-strand repair: Past and present. Exp Cell Res. 319:575–587. 2013. View Article : Google Scholar : PubMed/NCBI
42	Wu LC, Wang ZW, Tsan JT, Spillman MA, Phung A, Xu XL, Yang MC, Hwang LY, Bowcock AM and Baer R: Identification of a RING protein that can interact in vivo with the BRCA1 gene product. Nat Genet. 14:430–440. 1996. View Article : Google Scholar : PubMed/NCBI
43	Scully R, Chen J, Plug A, Xiao Y, Weaver D, Feunteun J, Ashley T and Livingston DM: Association of BRCA1 with Rad51 in mitotic and meiotic cells. Cell. 88:265–275. 1997. View Article : Google Scholar : PubMed/NCBI
44	Zhao W, Steinfeld JB, Liang F, Chen X, Maranon DG, Jian Ma C, Kwon Y, Rao T, Wang W, Sheng C, et al: BRCA1-BARD1 promotes RAD51-mediated homologous DNA pairing. Nature. 550:360–365. 2017. View Article : Google Scholar : PubMed/NCBI
45	Mani C, Jonnalagadda S, Lingareddy J, Awasthi S, Gmeiner WH and Palle K: Prexasertib treatment induces homologous recombination deficiency and synergizes with olaparib in triple-negative breast cancer cells. Breast Cancer Res. 21:1042019. View Article : Google Scholar : PubMed/NCBI
46	Lei H, Jin J, Liu M, Li X, Luo H, Yang L, Xu H and Wu Y: Chk1 inhibitors overcome imatinib resistance in chronic myeloid leukemia cells. Leukemia Res. 64:17–23. 2018. View Article : Google Scholar