Deficiency of 53BP1 inhibits the radiosensitivity of colorectal cancer

Xiao,Yong; Zheng,Xiumei; Huang,Ai; Liu,Tao; Zhang,Tao; Ma,Hong

doi:10.3892/ijo.2016.3629

Deficiency of 53BP1 inhibits the radiosensitivity of colorectal cancer

Authors:
- Yong Xiao
- Xiumei Zheng
- Ai Huang
- Tao Liu
- Tao Zhang
- Hong Ma
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Affiliations: Cancer Center, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China
Published online on: July 25, 2016 https://doi.org/10.3892/ijo.2016.3629
Pages: 1600-1608

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Abstract

The present study aimed to observe the influence of p53 binding protein 1 (53BP1) silencing on the radiosensitivity of colorectal cancer (CRC) cells and investigated the potential underlying mechanisms. The differences in radiosensitivity among four CRC cells were detected by the clone formation assay, while the expression of their 53BP1 was detected by the western blot analysis. HCT116 cells with relatively high expression of 53BP1 were selected to silence the expression of 53BP1 by shRNA intervention. The influence on proliferation, apoptosis, and cell cycle distribution was detected by immunofluorescent staining of Ki-67 and flow cytometry. The expression of relevant proteins in the apoptotic pathway ATM-CHK2-p53 was further analyzed by western blot analysis. The expression of 53BP1 was found to be closely related to the radiosensitivity of the CRC cells. Decreased expression of 53BP1 led to the tolerance of HCT116 cells to radiation. The detection of tumor proliferation, apoptosis, and cell cycle showed that decreased expression of 53BP1 resulted in an increased S-phase percentage of HCT116 cells, an increased proliferating rate, and a decreased apoptotic rate after radiation. The analysis of the molecular pathway showed that the reduced expression of 53BP1 decreased the protein expression of ATM, CHK2, and the phosphorylated products associated with the p53 apoptotic pathway. In conclusion, decreased expression of 53BP1 leads to radiotolerance of CRC cells, and the underlying mechanism is probably related to the decreased expression of relevant proteins in the ATM-CHK2-p53 pathway, which affects cell cycle, apoptosis and proliferation.

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1	Shin SJ, Yoon HI, Kim NK, Lee KY, Min BS, Ahn JB, Keum KC and Koom WS: Upfront systemic chemotherapy and preoperative short-course radiotherapy with delayed surgery for locally advanced rectal cancer with distant metastases. Radiat Oncol. 6:992011. View Article : Google Scholar : PubMed/NCBI
2	Zhang T, Liang ZW, Han J, Bi JP, Yang ZY and Ma H: Double-arc volumetric modulated therapy improves dose distribution compared to static gantry IMRT and 3D conformal radiotherapy for adjuvant therapy of gastric cancer. Radiat Oncol. 10:1142015. View Article : Google Scholar : PubMed/NCBI
3	Wang M, Kern AM, Hülskötter M, Greninger P, Singh A, Pan Y, Chowdhury D, Krause M, Baumann M, Benes CH, et al: EGFR-mediated chromatin condensation protects KRAS-mutant cancer cells against ionizing radiation. Cancer Res. 74:2825–2834. 2014. View Article : Google Scholar : PubMed/NCBI
4	Di Micco R, Fumagalli M, Cicalese A, Piccinin S, Gasparini P, Luise C, Schurra C, Garre’ M, Nuciforo PG, Bensimon A, et al: Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature. 444:638–642. 2006. View Article : Google Scholar : PubMed/NCBI
5	Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva N, Vassiliou LV, Kolettas E, Niforou K, Zoumpourlis VC, et al: Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature. 444:633–637. 2006. View Article : Google Scholar : PubMed/NCBI
6	Carr SM, Munro S, Zalmas LP, Fedorov O, Johansson C, Krojer T, Sagum CA, Bedford MT, Oppermann U and La Thangue NB: Lysine methylation-dependent binding of 53BP1 to the pRb tumor suppressor. Proc Natl Acad Sci USA. 111:11341–11346. 2014. View Article : Google Scholar : PubMed/NCBI
7	Bouwman P, Aly A, Escandell JM, Pieterse M, Bartkova J, van der Gulden H, Hiddingh S, Thanasoula M, Kulkarni A, Yang Q, et al: 53BP1 loss rescues BRCA1 deficiency and is associated with triple-negative and BRCA-mutated breast cancers. Nat Struct Mol Biol. 17:688–695. 2010. View Article : Google Scholar : PubMed/NCBI
8	Nuciforo PG, Luise C, Capra M, Pelosi G and d’Adda di Fagagna F: Complex engagement of DNA damage response pathways in human cancer and in lung tumor progression. Carcinogenesis. 28:2082–2088. 2007. View Article : Google Scholar : PubMed/NCBI
9	Neboori HJ, Haffty BG, Wu H, Yang Q, Aly A, Goyal S, Schiff D, Moran MS, Golhar R, Chen C, et al: Low p53 binding protein 1 (53BP1) expression is associated with increased local recurrence in breast cancer patients treated with breast-conserving surgery and radiotherapy. Int J Radiat Oncol Biol Phys. 83:e677–e683. 2012. View Article : Google Scholar : PubMed/NCBI
10	Squatrito M, Vanoli F, Schultz N, Jasin M and Holland EC: 53BP1 is a haploinsufficient tumor suppressor and protects cells from radiation response in glioma. Cancer Res. 72:5250–5260. 2012. View Article : Google Scholar : PubMed/NCBI
11	Ma H, Bi J, Liu T, Ke Y, Zhang S and Zhang T: Icotinib hydrochloride enhances the effect of radiotherapy by affecting DNA repair in colorectal cancer cells. Oncol Rep. 33:1161–1170. 2015.PubMed/NCBI
12	Shi Y, Felley-Bosco E, Marti TM, Orlowski K, Pruschy M and Stahel RA: Starvation-induced activation of ATM/Chk2/p53 signaling sensitizes cancer cells to cisplatin. BMC Cancer. 12:5712012. View Article : Google Scholar : PubMed/NCBI
13	Chung YM, Park SH, Tsai WB, Wang SY, Ikeda MA, Berek JS, Chen DJ and Hu MC: FOXO3 signalling links ATM to the p53 apoptotic pathway following DNA damage. Nat Commun. 3:10002012. View Article : Google Scholar : PubMed/NCBI
14	Bartkova J, Horejsí Z, Sehested M, Nesland JM, Rajpert-De Meyts E, Skakkebaek NE, Stucki M, Jackson S, Lukas J and Bartek J: DNA damage response mediators MDC1 and 53BP1: Constitutive activation and aberrant loss in breast and lung cancer, but not in testicular germ cell tumours. Oncogene. 26:7414–7422. 2007. View Article : Google Scholar : PubMed/NCBI
15	Morales JC, Franco S, Murphy MM, Bassing CH, Mills KD, Adams MM, Walsh NC, Manis JP, Rassidakis GZ, Alt FW, et al: 53BP1 and p53 synergize to suppress genomic instability and lymphomagenesis. Proc Natl Acad Sci USA. 103:3310–3315. 2006. View Article : Google Scholar : PubMed/NCBI
16	Ausborn NL, Wang T, Wentz SC, Washington MK, Merchant NB, Zhao Z, Shyr Y, Chakravarthy AB and Xia F: 53BP1 expression is a modifier of the prognostic value of lymph node ratio and CA 19-9 in pancreatic adenocarcinoma. BMC Cancer. 13:1552013. View Article : Google Scholar : PubMed/NCBI
17	Squatrito M, Brennan CW, Helmy K, Huse JT, Petrini JH and Holland EC: Loss of ATM/Chk2/p53 pathway components accelerates tumor development and contributes to radiation resistance in gliomas. Cancer Cell. 18:619–629. 2010. View Article : Google Scholar : PubMed/NCBI
18	Ward IM, Difilippantonio S, Minn K, Mueller MD, Molina JR, Yu X, Frisk CS, Ried T, Nussenzweig A and Chen J: 53BP1 cooperates with p53 and functions as a haploinsufficient tumor suppressor in mice. Mol Cell Biol. 25:10079–10086. 2005. View Article : Google Scholar : PubMed/NCBI
19	Gorgoulis VG, Vassiliou LV, Karakaidos P, Zacharatos P, Kotsinas A, Liloglou T, Venere M, Ditullio RA Jr, Kastrinakis NG, Levy B, et al: Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature. 434:907–913. 2005. View Article : Google Scholar : PubMed/NCBI
20	Khanna A: DNA damage in cancer therapeutics: A boon or a curse? Cancer Res. 75:2133–2138. 2015. View Article : Google Scholar : PubMed/NCBI
21	Karanika S, Karantanos T, Li L, Corn PG and Thompson TC: DNA damage response and prostate cancer: Defects, regulation and therapeutic implications. Oncogene. 34:2815–2822. 2015. View Article : Google Scholar :
22	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
23	Lai TC, Chow KC, Lin TY, Chiang IP, Fang HY, Chen CY and Ho SP: Expression of 53BP1 as a cisplatin-resistant marker in patients with lung adenocarcinomas. Oncol Rep. 24:321–328. 2010.PubMed/NCBI
24	Li X, Kong X, Kong X, Wang Y, Yan S and Yang Q: 53BP1 sensitizes breast cancer cells to 5-fluorouracil. PLoS One. 8:e749282013. View Article : Google Scholar : PubMed/NCBI
25	Clarke AR, Jones N, Pryde F, Adachi Y and Sansom OJ: 53BP1 deficiency in intestinal enterocytes does not alter the immediate response to ionizing radiation, but leads to increased nuclear area consistent with polyploidy. Oncogene. 26:6349–6355. 2007. View Article : Google Scholar : PubMed/NCBI
26	Cao L, Xu X, Bunting SF, Liu J, Wang RH, Cao LL, Wu JJ, Peng TN, Chen J, Nussenzweig A, et al: A selective requirement for 53BP1 in the biological response to genomic instability induced by Brca1 deficiency. Mol Cell. 35:534–541. 2009. View Article : Google Scholar : PubMed/NCBI
27	Han X, Zhang L, Chung J, Mayca Pozo F, Tran A, Seachrist DD, Jacobberger JW, Keri RA, Gilmore H and Zhang Y: UbcH7 regulates 53BP1 stability and DSB repair. Proc Natl Acad Sci USA. 111:17456–17461. 2014. View Article : Google Scholar : PubMed/NCBI
28	Bi J, Huang A, Liu T, Zhang T and Ma H: Expression of DNA damage checkpoint 53BP1 is correlated with prognosis, cell proliferation and apoptosis in colorectal cancer. Int J Clin Exp Pathol. 8:6070–6082. 2015.PubMed/NCBI
29	Sun CC, Chiu HT, Lin YF, Lee KY and Pang JH: Y-27632, a ROCK inhibitor, promoted limbal epithelial cell proliferation and corneal wound healing. PLoS One. 10:e01445712015. View Article : Google Scholar : PubMed/NCBI
30	Li T, Shi HY, Hua YX, Gao C, Xia Q, Yang G and Li B: Effects of allicin on the proliferation and cell cycle of chondrocytes. Int J Clin Exp Pathol. 8:12525–12532. 2015.