CDK5RAP1 deficiency induces cell cycle arrest and apoptosis in human breast cancer cell line by the ROS/JNK signaling pathway

Wang,Hongbin; Wei,Li; Li,Changli; Zhou,Jing; Li,Zhigao

doi:10.3892/or.2015.3736

CDK5RAP1 deficiency induces cell cycle arrest and apoptosis in human breast cancer cell line by the ROS/JNK signaling pathway

Authors:
- Hongbin Wang
- Li Wei
- Changli Li
- Jing Zhou
- Zhigao Li
View Affiliations

Affiliations: Department of Breast Surgery, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang 150081, P.R. China
Published online on: January 20, 2015 https://doi.org/10.3892/or.2015.3736
Pages: 1089-1096

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

Cyclin-dependent kinase 5 regulatory subunit associated protein 1 (CDK5RAP1) is an enzyme which post-synthetically converts the RNA modification N6-iso-pentenyladenosine (i6A) into 2-methylthio-N6-isopentenyladenosine (ms2i6A). However, the interaction between CDK5RAP1 deficiency and cell apoptosis has not been studied. Breast cancer has long been a leading cause of mortality in the world. Therefore, in the present study, CDK5RAP1 deficiency in a human breast cancer cell line was investigated. CDK5RAP1 small interfering RNA (siRNA) and negative control siRNA were transfected into MCF-7 cells, and the cells were further incubated for 48 h. CDK5RAP1 deficiency suppressed tumor growth in MCF-7 cells and arrested the cells at G2/M phase. CDK5RAP1 deficiency also induced cell apoptosis and reactive oxygen species (ROS) generation. Furthermore, western blot analysis showed that the expression of phospho-c-Jun N-terminal kinase (p-JNK), p53, caspase-9 and caspase-3 were upregulated in CDK5RAP1-deficient MCF-7 cells. Pretreatment with N-acetyl-cysteine (NAC), the inhibitor of ROS, or with SP600125, the inhibitor of JNK, prevented the apoptosis and the high expression of p-JNK, p53, caspase-9 and caspase-3 in CDK5RAP1-deficient MCF-7 cells. Taken together, these data indicated that CDK5RAP1 deficiency induced cell cycle arrest and apoptosis in human breast cancer MCF-7 cells by the ROS/JNK signaling pathway. Our findings indicated a novel therapeutic strategy for cancer.

View Figures

View References

1	Atta M, Mulliez E, Arragain S, Forouhar F, Hunt JF and Fontecave M: S-Adenosylmethionine-dependent radical-based modification of biological macromolecules. Curr Opin Struct Biol. 20:684–692. 2010. View Article : Google Scholar : PubMed/NCBI
2	Kaminska KH, Baraniak U, Boniecki M, Nowaczyk K, Czerwoniec A and Bujnicki JM: Structural bioinformatics analysis of enzymes involved in the biosynthesis pathway of the hypermodified nucleoside ms(²)io(⁶)A37 in tRNA. Proteins. 70:1–18. 2008. View Article : Google Scholar
3	Pierrel F, Douki T, Fontecave M and Atta M: MiaB protein is a bifunctional radical-S-adenosylmethionine enzyme involved in thiolation and methylation of tRNA. J Biol Chem. 279:47555–47563. 2004. View Article : Google Scholar : PubMed/NCBI
4	Bouadloun F, Srichaiyo T, Isaksson LA and Bjork GR: Influence of modification next to the anticodon in tRNA on codon context sensitivity of translational suppression and accuracy. J Bacteriol. 166:1022–1027. 1986.PubMed/NCBI
5	Jenner LB, Demeshkina N, Yusupova G and Yusupov M: Structural aspects of messenger RNA reading frame maintenance by the ribosome. Nat Struct Mol Biol. 17:555–560. 2010. View Article : Google Scholar : PubMed/NCBI
6	Gong CX and Iqbal K: Hyperphosphorylation of microtubule-associated protein tau: a promising therapeutic target for Alzheimer disease. Curr Med Chem. 15:2321–2328. 2008. View Article : Google Scholar : PubMed/NCBI
7	Esberg B, Leung HC, Tsui HC, Bjork GR and Winkler ME: Identification of the miaB gene, involved in methylthiolation of isopentenylated A37 derivatives in the tRNA of Salmonella typhimurium and Escherichia coli. J Bacteriol. 181:7256–7265. 1999.PubMed/NCBI
8	Globisch D, Pearson D, Hienzsch A, Brückl T, Wagner M, Thoma I, Thumbs P, Reiter V, Kneuttinger AC, Müller M, Sieber SA and Carell T: Systems-based analysis of modified tRNA bases. Angew Chem Int Ed Engl. 50:9739–9742. 2011. View Article : Google Scholar : PubMed/NCBI
9	Brody JG, Rudel RA, Michels KB, Moysich KB, Bernstein L, Attfield KR and Gray S: Environmental pollutants, diet, physical activity, body size, and breast cancer: where do we stand in research to identify opportunities for prevention? Cancer. 109:2627–2634. 2007. View Article : Google Scholar : PubMed/NCBI
10	Fulda S and Debatin KM: Targeting apoptosis pathways in cancer therapy. Curr Cancer Drug Targets. 4:569–576. 2004. View Article : Google Scholar : PubMed/NCBI
11	Gamet-Payrastre L, Li P, Lumeau S, Cassar G, Dupont MA, Chevolleau S, Gasc N, Tulliez J and Tercé F: Sulforaphane, a naturally occurring isothiocyanate, induces cell cycle arrest and apoptosis in HT29 human colon cancer cells. Cancer Res. 60:1426–1433. 2000.PubMed/NCBI
12	Snoek BC, de Wilt LH, Jansen G and Peters GJ: Role of E3 ubiquitin ligases in lung cancer. World J Clin Oncol. 4:58–69. 2013. View Article : Google Scholar : PubMed/NCBI
13	Myatt SS, Brosens JJ and Lam EW: Sense and sensitivity: FOXO and ROS in cancer development and treatment. Antioxid Redox Signal. 14:675–687. 2011. View Article : Google Scholar
14	Trachootham D, Alexandre J and Huang P: Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov. 8:579–591. 2009. View Article : Google Scholar : PubMed/NCBI
15	Kong D, Zheng T, Zhang M, Wang D, Du S, Li X, Fang J and Cao X: Static mechanical stress induces apoptosis in rat endplate chondrocytes through MAPK and mitochondria-dependent caspase activation signaling pathways. PLoS One. 8:e694032013. View Article : Google Scholar : PubMed/NCBI
16	Uchakina ON, Ban H and McKallip RJ: Targeting hyaluronic acid production for the treatment of leukemia: treatment with 4-methylumbelliferone leads to induction of MAPK-mediated apoptosis in K562 leukemia. Leuk Res. 37:1294–1301. 2013. View Article : Google Scholar : PubMed/NCBI
17	Leber B, Geng F, Kale J and Andrews DW: Drugs targeting Bcl-2 family members as an emerging strategy in cancer. Expert Rev Mol Med. 12:e282010. View Article : Google Scholar : PubMed/NCBI
18	Yuan Z, Feng W, Hong J, Zheng Q, Shuai J and Ge Y: p38MAPK and ERK promote nitric oxide production in cultured human retinal pigmented epithelial cells induced by high concentration glucose. Nitric Oxide. 20:9–15. 2009. View Article : Google Scholar
19	Pozarowski P and Darzynkiewicz Z: Analysis of cell cycle by flow cytometry. Methods Mol Biol. 281:301–311. 2004.PubMed/NCBI
20	Rastogi RP, Singh SP, Häder DP and Sinha RP: Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2′, 7′-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 793. Biochem Biophys Res Commun. 397:603–607. 2010. View Article : Google Scholar : PubMed/NCBI
21	Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI
22	Kyhse-Andersen J: Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 10:203–209. 1984. View Article : Google Scholar : PubMed/NCBI
23	Lan T, Wang L, Xu Q, Liu W, Jin H, Mao W and Wang X and Wang X: Growth inhibitory effect of Cucurbitacin E on breast cancer cells. Int J Clin Exp Pathol. 6:1799–1805. 2013.PubMed/NCBI
24	Drexler HG: Review of alterations of the cyclin-dependent kinase inhibitor INK4 family genes p15, p16, p18 and p19 in human leukemia-lymphoma cells. Leukemia. 12:845–859. 1998. View Article : Google Scholar : PubMed/NCBI
25	Padua MB and Hansen PJ: Changes in expression of cell-cycle-related genes in PC-3 prostate cancer cells caused by ovine uterine serpin. J Cell Biochem. 107:1182–1188. 2009. View Article : Google Scholar : PubMed/NCBI
26	Wyllie AH, Kerr JF and Currie AR: Cell death: the significance of apoptosis. Int Rev Cytol. 68:251–306. 1980. View Article : Google Scholar : PubMed/NCBI
27	Cooper WA, Kohonen-Corish MR, Zhuang L, McCaughan B, Kennedy C, Screaton G, Sutherland RL and Lee CS: Role and prognostic significance of tumor necrosis factor-related apoptosis-inducing ligand death receptor DR5 in nonsmall-cell lung cancer and precursor lesions. Cancer. 113:135–142. 2008. View Article : Google Scholar : PubMed/NCBI
28	Dewaele M, Maes H and Agostinis P: ROS-mediated mechanisms of autophagy stimulation and their relevance in cancer therapy. Autophagy. 6:838–854. 2010. View Article : Google Scholar : PubMed/NCBI
29	Madureira PA, Hill R, Miller VA, Giacomantonio C, Lee PW and Waisman DM: Annexin A2 is a novel cellular redox regulatory protein involved in tumorigenesis. Oncotarget. 2:1075–1093. 2011.PubMed/NCBI
30	Kirshner JR, He S, Balasubramanyam V, Kepros J, Yang CY, Zhang M, Du Z, Barsoum J and Bertin J: Elesclomol induces cancer cell apoptosis through oxidative stress. Mol Cancer Ther. 7:2319–2327. 2008. View Article : Google Scholar : PubMed/NCBI
31	You BR and Park WH: Zebularine-induced apoptosis in Calu-6 lung cancer cells is influenced by ROS and GSH level changes. Tumour Biol. 34:1145–1153. 2013. View Article : Google Scholar : PubMed/NCBI
32	Strickertsson JA, Desler C, Martin-Bertelsen T, Machado AM, Wadstrøm T, Winther O, Rasmussen LJ and Friis-Hansen L: Enterococcus faecalis infection causes inflammation, intra-cellular oxphos-independent ROS production, and DNA damage in human gastric cancer cells. PLoS One. 8:e631472013. View Article : Google Scholar
33	Lee SJ, Kim MS, Park JY, Woo JS and Kim YK: 15-Deoxy-delta 12,14-prostaglandin J2 induces apoptosis via JNK-mediated mitochondrial pathway in osteoblastic cells. Toxicology. 248:121–129. 2008. View Article : Google Scholar : PubMed/NCBI
34	Lane DP: Cancer. p53, guardian of the genome. Nature. 358:15–16. 1992. View Article : Google Scholar : PubMed/NCBI
35	Wang Q, Su L, Liu N, Zhang L, Xu W and Fang H: Cyclin dependent kinase 1 inhibitors: a review of recent progress. Curr Med Chem. 18:2025–2043. 2011. View Article : Google Scholar : PubMed/NCBI
36	Lee K, Hart MR, Briehl MM, Mazar AP and Tome ME: The copper chelator ATN-224 induces caspase-independent cell death in diffuse large B cell lymphoma. Int J Oncol. 45:439–447. 2014.PubMed/NCBI