Enhancement of radiosensitivity by inhibition of c-Jun N-terminal kinase activity in a Lewis lung carcinoma‑bearing subcutaneous tumor mouse model

Li,Chun-Hao; Lim,Sa-Hoe; Ryu,Hyang-Hwa; Moon,Kyung‑Sub; Jung,Tae-Young; Jung,Shin

doi:10.3892/or.2016.5204

Enhancement of radiosensitivity by inhibition of c-Jun N-terminal kinase activity in a Lewis lung carcinoma‑bearing subcutaneous tumor mouse model

Authors:
- Chun-Hao Li
- Sa-Hoe Lim
- Hyang-Hwa Ryu
- Kyung‑Sub Moon
- Tae-Young Jung
- Shin Jung
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Affiliations: Brain Tumor Research Laboratory, Chonnam National University Research Institute of Medical Sciences, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Jeollanam-do 519-763, Republic of Korea, Department of Neurosurgery, Chonnam National University Medical School and Chonnam National University Hwasun Hospital, Hwasun, Jeollanam-do 519-763, Republic of Korea
Published online on: October 25, 2016 https://doi.org/10.3892/or.2016.5204
Pages: 3397-3404

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Abstract

Stereotactic radiosurgery has been recognized as an effective treatment approach for metastatic brain tumors. By increasing the sensitivity of the tumor to radiation and decreasing the marginal dose, it is possible to improve therapeutic efficacy and decrease side-effects. In radiation-induced cells, c-Jun N-terminal kinase (JNK) signaling mediates the phosphorylation of H2AX, which indicates DNA damage sensitivity and modulates the effect of radiation. Lewis lung cancer (LLC) and breast cancer (4T1) cells were irradiated with a Gamma Knife in cell culture tubes. To evaluate the relationship between radiosensitivity and JNK activity, clonogenic assay was performed. DNA damage response was estimated by γH2AX focus formation assay and apoptosis‑related protein levels were assessed by western blotting. The mice were subcutaneously inoculated with LLC cells, and irradiated concomitantly with JNK inhibitor treatment. The effect of the JNK inhibitor was investigated by tumor volumetry and immunohistochemistry. γH2AX expression, which mediates repair of radiation‑induced DNA damage, was reduced in the cancer cell group pretreated with the JNK inhibitor. This finding shows that JNK inhibition may increase the radiosensitivity in radiated lung and breast cancer cells. For the in vivo study, irradiated tumor growth was significantly delayed in the JNK inhibitor-treated mouse group. Blockade of JNK signaling decreased γH2AX expression and increased apoptosis in the radiation-induced cancer cells. JNK inhibitor may be useful for enhancing the radiosensitivity of lung and breast cancer cells and improving the treatment efficacy of radiosurgical approaches for metastatic brain tumors.

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1	Davis RJ: Signal transduction by the JNK group of MAP kinases. Cell. 103:239–252. 2000. View Article : Google Scholar : PubMed/NCBI
2	Bode AM and Dong Z: The functional contrariety of JNK. Mol Carcinog. 46:591–598. 2007. View Article : Google Scholar : PubMed/NCBI
3	Weston CR and Davis RJ: The JNK signal transduction pathway. Curr Opin Cell Biol. 19:142–149. 2007. View Article : Google Scholar : PubMed/NCBI
4	Lu C, Zhu F, Cho YY, Tang F, Zykova T, Ma WY, Bode AM and Dong Z: Cell apoptosis: Requirement of H2AX in DNA ladder formation, but not for the activation of caspase-3. Mol Cell. 23:121–132. 2006. View Article : Google Scholar : PubMed/NCBI
5	Yue WY, Clark JJ, Telisak M and Hansen MR: Inhibition of c-Jun N-terminal kinase activity enhances vestibular schwannoma cell sensitivity to gamma irradiation. Neurosurgery. 73:506–516. 2013. View Article : Google Scholar : PubMed/NCBI
6	Bennett BL, Sasaki DT, Murray BW, O'Leary EC, Sakata ST, Xu W, Leisten JC, Motiwala A, Pierce S, Satoh Y, et al: SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase. Proc Natl Acad Sci USA. 98:13681–13686. 2001. View Article : Google Scholar : PubMed/NCBI
7	Guan QH, Pei DS, Zhang QG, Hao ZB, Xu TL and Zhang GY: The neuroprotective action of SP600125, a new inhibitor of JNK, on transient brain ischemia/reperfusion-induced neuronal death in rat hippocampal CA1 via nuclear and non-nuclear pathways. Brain Res. 1035:51–59. 2005. View Article : Google Scholar : PubMed/NCBI
8	Li S, Li C, Ryu HH, Lim SH, Jang WY and Jung S: Bacitracin inhibits the migration of U87-MG glioma cells via interferences of the integrin outside-in signaling pathway. J Korean Neurosurg Soc. 59:106–116. 2016. View Article : Google Scholar : PubMed/NCBI
9	Jin SG, Ryu HH, Li SY, Li CH, Lim SH, Jang WY and Jung S: Nogo-A inhibits the migration and invasion of human malignant glioma U87MG cells. Oncol Rep. 35:3395–3402. 2016.PubMed/NCBI
10	Wen M, Jung S, Moon KS, Jiang SN, Li SY and Min JJ: Targeting orthotopic glioma in mice with genetically engineered Salmonella typhimurium. J Korean Neurosurg Soc. 55:131–135. 2014. View Article : Google Scholar : PubMed/NCBI
11	Wang W, Shi L, Xie Y, Ma C, Li W, Su X, Huang S, Chen R, Zhu Z, Mao Z, et al: SP600125, a new JNK inhibitor, protects dopaminergic neurons in the MPTP model of Parkinson's disease. Neurosci Res. 48:195–202. 2004. View Article : Google Scholar : PubMed/NCBI
12	Yue WY, Clark JJ, Fernando A, Domann F and Hansen MR: Contribution of persistent C-Jun N-terminal kinase activity to the survival of human vestibular schwannoma cells by suppression of accumulation of mitochondrial superoxides. Neuro Oncol. 13:961–973. 2011. View Article : Google Scholar : PubMed/NCBI
13	Lin NU, Bellon JR and Winer EP: CNS metastases in breast cancer. J Clin Oncol. 22:3608–3617. 2004. View Article : Google Scholar : PubMed/NCBI
14	Shaw MG and Ball DL: Treatment of brain metastases in lung cancer: Strategies to avoid/reduce late complications of whole brain radiation therapy. Curr Treat Options Oncol. 14:553–567. 2013. View Article : Google Scholar : PubMed/NCBI
15	Jackson SP and Bartek J: The DNA-damage response in human biology and disease. Nature. 461:1071–1078. 2009. View Article : Google Scholar : PubMed/NCBI
16	Fernandez-Capetillo O, Chen HT, Celeste A, Ward I, Romanienko PJ, Morales JC, Naka K, Xia Z, Camerini-Otero RD, Motoyama N, et al: DNA damage-induced G₂-M checkpoint activation by histone H2AX and 53BP1. Nat Cell Biol. 4:993–997. 2002. View Article : Google Scholar : PubMed/NCBI
17	Paull TT, Rogakou EP, Yamazaki V, Kirchgessner CU, Gellert M and Bonner WM: A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage. Curr Biol. 10:886–895. 2000. View Article : Google Scholar : PubMed/NCBI
18	Burma S, Chen BP, Murphy M, Kurimasa A and Chen DJ: ATM phosphorylates histone H2AX in response to DNA double-strand breaks. J Biol Chem. 276:42462–42467. 2001. View Article : Google Scholar : PubMed/NCBI
19	Wang H, Wang M, Wang H, Böcker W and Iliakis G: Complex H2AX phosphorylation patterns by multiple kinases including ATM and DNA-PK in human cells exposed to ionizing radiation and treated with kinase inhibitors. J Cell Physiol. 202:492–502. 2005. View Article : Google Scholar : PubMed/NCBI
20	Ward IM and Chen J: Histone H2AX is phosphorylated in an ATR-dependent manner in response to replicational stress. J Biol Chem. 276:47759–47762. 2001.PubMed/NCBI
21	Hibi M, Lin A, Smeal T, Minden A and Karin M: Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev. 7:2135–2148. 1993. View Article : Google Scholar : PubMed/NCBI
22	Waetzig V and Herdegen T: Context-specific inhibition of JNKs: Overcoming the dilemma of protection and damage. Trends Pharmacol Sci. 26:455–461. 2005.PubMed/NCBI
23	Oltmanns U, Issa R, Sukkar MB, John M and Chung KF: Role of c-jun N-terminal kinase in the induced release of GM-CSF, RANTES and IL-8 from human airway smooth muscle cells. Br J Pharmacol. 139:1228–1234. 2003. View Article : Google Scholar : PubMed/NCBI
24	Barr RK, Kendrick TS and Bogoyevitch MA: Identification of the critical features of a small peptide inhibitor of JNK activity. J Biol Chem. 277:10987–10997. 2002. View Article : Google Scholar : PubMed/NCBI
25	An J, Huang YC, Xu QZ, Zhou LJ, Shang ZF, Huang B, Wang Y, Liu XD, Wu DC and Zhou PK: DNA-PKcs plays a dominant role in the regulation of H2AX phosphorylation in response to DNA damage and cell cycle progression. BMC Mol Biol. 11:182010. View Article : Google Scholar : PubMed/NCBI
26	Kim CH, Won M, Choi CH, Ahn J, Kim BK, Song KB, Kang CM and Chung KS: Increase of RhoB in gamma-radiation-induced apoptosis is regulated by c-Jun N-terminal kinase in Jurkat T cells. Biochem Biophys Res Commun. 391:1182–1186. 2010. View Article : Google Scholar : PubMed/NCBI
27	Han Y, Wang Y, Xu HT, Yang LH, Wei Q, Liu Y, Zhang Y, Zhao Y, Dai SD, Miao Y, et al: X-radiation induces non-small-cell lung cancer apoptosis by upregulation of Axin expression. Int J Radiat Oncol Biol Phys. 75:518–526. 2009. View Article : Google Scholar : PubMed/NCBI
28	Gao Z, Sarsour EH, Kalen AL, Li L, Kumar MG and Goswami PC: Late ROS accumulation and radiosensitivity in SOD1-overexpressing human glioma cells. Free Radic Biol Med. 45:1501–1509. 2008. View Article : Google Scholar : PubMed/NCBI
29	Shen HM and Liu ZG: JNK signaling pathway is a key modulator in cell death mediated by reactive oxygen and nitrogen species. Free Radic Biol Med. 40:928–939. 2006. View Article : Google Scholar : PubMed/NCBI
30	Jena NR: DNA damage by reactive species: Mechanisms, mutation and repair. J Biosci. 37:503–517. 2012. View Article : Google Scholar : PubMed/NCBI
31	Yu CC, Ko FY, Yu CS, Lin CC, Huang YP, Yang JS, Lin JP and Chung JG: Norcantharidin triggers cell death and DNA damage through S-phase arrest and ROS-modulated apoptotic pathways in TSGH 8301 human urinary bladder carcinoma cells. Int J Oncol. 41:1050–1060. 2012.PubMed/NCBI
32	Zeng H, Kong X, Peng H, Chen Y, Cai S, Luo H and Chen P: Apoptosis and Bcl-2 family proteins, taken to chronic obstructive pulmonary disease. Eur Rev Med Pharmacol Sci. 16:711–727. 2012.PubMed/NCBI
33	Tang S, Hu J, Meng Q, Dong X, Wang K, Qi Y, Chu C, Zhang X and Hou L: Daidzein induced apoptosis via down-regulation of Bcl-2/Bax and triggering of the mitochondrial pathway in BGC-823 cells. Cell Biochem Biophys. 65:197–202. 2013. View Article : Google Scholar : PubMed/NCBI
34	Miwa M and Masutani M: PolyADP-ribosylation and cancer. Cancer Sci. 98:1528–1535. 2007. View Article : Google Scholar : PubMed/NCBI
35	Wei L, Nakajima S, Hsieh CL, Kanno S, Masutani M, Levine AS, Yasui A and Lan L: Damage response of XRCC1 at sites of DNA single strand breaks is regulated by phosphorylation and ubiquitylation after degradation of poly(ADP-ribose). J Cell Sci. 126:4414–4423. 2013. View Article : Google Scholar : PubMed/NCBI
36	Germain M, Affar EB, D'Amours D, Dixit VM, Salvesen GS and Poirier GG: Cleavage of automodified poly(ADP-ribose) polymerase during apoptosis. Evidence for involvement of caspase-7. J Biol Chem. 274:28379–28384. 1999. View Article : Google Scholar : PubMed/NCBI