MELK as a potential target to control cell proliferation in triple‑negative breast cancer MDA‑MB‑231 cells

Li,Gang; Yang,Mei; Zuo,Li; Wang,Mei‑Xing

doi:10.3892/ol.2018.8543

MELK as a potential target to control cell proliferation in triple‑negative breast cancer MDA‑MB‑231 cells

Authors:
- Gang Li
- Mei Yang
- Li Zuo
- Mei‑Xing Wang
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Affiliations: Department of Medical Oncology, Branch of Minhang, Fudan University Shanghai Cancer Center, Shanghai 200240, P.R. China
Published online on: April 20, 2018 https://doi.org/10.3892/ol.2018.8543
Pages: 9934-9940

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Abstract

Maternal embryonic leucine zipper kinase (MELK) is an important regulator in tumorigenesis of human breast cancer, and if silenced leads to programmed cell death in specific breast cancer cell lines, including MDA‑MB‑231 cells. In the present study, RNA interference, proliferation assay and semi‑quantification of cell cycle relative proteins were performed to determine the effects of MELK in human breast cancer cells. Data demonstrated that the highest level of MELK protein in the MDA‑MB‑231 cell line among eight breast cancer cell lines. The sensitivity of MELK small interfering‑RNA varied in different breast cancer cell lines, but MELK silencing resulted in marked suppression of proliferation of triple‑negative breast cancer (TNBC) and non‑TNBC cells. Specific silencing of MELK caused G2 arrest in TNBC MDA‑MB‑231 and HCC1143 cells, and G1 arrest in non‑TNBC T47D and MCF7 cells. Notably, the knockdown of MELK did not induce apoptosis in HCC1143 cells, indicated by the lack of caspase‑3 expression. In addition, in response to MELK silencing, cyclin B and cyclin D1 were downregulated in four breast cancer cell lines. Furthermore, the silencing of MELK resulted in the upregulation of p21, p27 and phosphorylated (p)‑c‑Jun N‑terminal kinase (JNK) in HCC1143 TNBC cells, and downregulation of p21 and p‑JNK in T47D non‑TNBC cells. Additionally, MELK protein was markedly suppressed in non‑TNBC cells in response to estrogen deprivation. The findings from the present study suggested that MELK may be a potential target in MDA‑MB‑231 cells, although genetic knockdown of MELK resulted in inhibitory effects on proliferation of TNBC and non‑TNBC cells. MELK exert its effect on different breast cancer cells via arrest of different cell cycle phases and therefore mediated by different mediators, which may be involved in the crosstalk with MELK signaling and with the estrogen receptor signaling pathway.

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1	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
2	Perou CM, Sørlie T, Eisen MB, van de Rijn M, Jeffrey SS, Rees CA, Pollack JR, Ross DT, Johnsen H, Akslen LA, et al: Molecular portraits of human breast tumours. Nature. 406:747–752. 2000. View Article : Google Scholar : PubMed/NCBI
3	Sørlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H, Hastie T, Eisen MB, van de Rijn M, Jeffrey SS, et al: Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci USA. 98:10869–10874. 2001. View Article : Google Scholar : PubMed/NCBI
4	Rakha EA, Reis-Filho JS and Ellis IO: Basal-like breast cancer: A critical review. J Clin Oncol. 26:2568–2581. 2008. View Article : Google Scholar : PubMed/NCBI
5	Jamdade VS, Sethi N, Mundhe NA, Kumar P, Lahkar M and Sinha N: Therapeutic targets of triple-negative breast cancer: A review. Br J Pharmacol. 172:4228–4237. 2015. View Article : Google Scholar : PubMed/NCBI
6	Soady KJ, Kendrick H, Gao Q, Tutt A, Zvelebil M, Ordonez LD, Quist J, Tan DW, Isacke CM, Grigoriadis A and Smalley MJ: Mouse mammary stem cells express prognostic markers for triple-negative breast cancer. Breast Cancer Res. 17:312015. View Article : Google Scholar : PubMed/NCBI
7	Chiotaki R, Polioudaki H and Theodoropoulos PA: Stem cell technology in breast cancer: Current status and potential applications. Stem Cells Cloning. 9:17–29. 2016.PubMed/NCBI
8	Jiang P and Zhang D: Maternal embryonic leucine zipper kinase (MELK): A novel regulator in cell cycle control, embryonic development, and cancer. Int J Mol Sci. 14:21551–21560. 2013. View Article : Google Scholar : PubMed/NCBI
9	Janostiak R, Rauniyar N, Lam TT, Ou J, Zhu LJ, Green MR and Wajapeyee N: MELK promotes melanoma growth by stimulating the NF-κB pathway. Cell Rep. 21:2829–2841. 2017. View Article : Google Scholar : PubMed/NCBI
10	Nakano I, Masterman-Smith M, Saigusa K, Paucar AA, Horvath S, Shoemaker L, Watanabe M, Negro A, Bajpai R, Howes A, et al: Maternal embryonic leucine zipper kinase is a key regulator of the proliferation of malignant brain tumors, including brain tumor stem cells. J Neurosci Res. 86:48–60. 2008. View Article : Google Scholar : PubMed/NCBI
11	Yoshimoto K, Ma X, Guan Y, Mizoguchi M, Nakamizo A, Amano T, Hata N, Kuga D and Sasaki T: Expression of stem cell marker and receptor kinase genes in glioblastoma tissue quantified by real-time RT-PCR. Brain Tumor Pathol. 28:291–296. 2011. View Article : Google Scholar : PubMed/NCBI
12	Nakano I, Paucar AA, Bajpai R, Dougherty JD, Zewail A, Kelly TK, Kim KJ, Ou J, Groszer M, Imura T, et al: Maternal embryonic leucine zipper kinase (MELK) regulates multipotent neural progenitor proliferation. J Cell Biol. 170:413–427. 2005. View Article : Google Scholar : PubMed/NCBI
13	Cancer Genome Atlas Network: Comprehensive molecular portraits of human breast tumours. Nature. 490:61–70. 2012. View Article : Google Scholar : PubMed/NCBI
14	Ma XJ, Dahiya S, Richardson E, Erlander M and Sgroi DC: Gene expression profiling of the tumor microenvironment during breast cancer progression. Breast Cancer Res. 11:R72009. View Article : Google Scholar : PubMed/NCBI
15	Richardson AL, Wang ZC, De Nicolo A, Lu X, Brown M, Miron A, Liao X, Iglehart JD, Livingston DM and Ganesan S: X chromosomal abnormalities in basal-like human breast cancer. Cancer Cell. 9:121–132. 2006. View Article : Google Scholar : PubMed/NCBI
16	Desmedt C, Piette F, Loi S, Wang Y, Lallemand F, Haibe-Kains B, Viale G, Delorenzi M, Zhang Y, d'Assignies MS, et al: Strong time dependence of the 76-gene prognostic signature for node-negative breast cancer patients in the TRANSBIG multicenter independent validation series. Clin Cancer Res. 13:3207–3214. 2007. View Article : Google Scholar : PubMed/NCBI
17	Hatzis C, Pusztai L, Valero V, Booser DJ, Esserman L, Lluch A, Vidaurre T, Holmes F, Souchon E, Wang H, et al: A genomic predictor of response and survival following taxane-anthracycline chemotherapy for invasive breast cancer. JAMA. 305:1873–1881. 2011. View Article : Google Scholar : PubMed/NCBI
18	Schmidt M, Böhm D, von Törne C, Steiner E, Puhl A, Pilch H, Lehr HA, Hengstler JG, Kölbl H and Gehrmann M: The humoral immune system has a key prognostic impact in node-negative breast cancer. Cancer Res. 68:5405–5413. 2008. View Article : Google Scholar : PubMed/NCBI
19	Wang Y, Klijn JG, Zhang Y, Sieuwerts AM, Look MP, Yang F, Talantov D, Timmermans M, Meijer-van Gelder ME, Yu J, et al: Gene-expression profiles to predict distant metastasis of lymph-node-negative primary breast cancer. Lancet. 365:671–679. 2005. View Article : Google Scholar : PubMed/NCBI
20	Esserman LJ, Berry DA, Cheang MC, Yau C, Perou CM, Carey L, DeMichele A, Gray JW, Conway-Dorsey K, Lenburg ME, et al: Chemotherapy response and recurrence-free survival in neoadjuvant breast cancer depends on biomarker profiles: Results from the I-SPY 1 TRIAL (CALGB 150007/150012; ACRIN 6657). Breast Cancer Res Treat. 132:1049–1062. 2012. View Article : Google Scholar : PubMed/NCBI
21	Wang Y, Lee YM, Baitsch L, Huang A, Xiang Y, Tong H, Lako A, Von T, Choi C, Lim E, et al: MELK is an oncogenic kinase essential for mitotic progression in basal-like breast cancer cells. Elife. 3:e017632014. View Article : Google Scholar : PubMed/NCBI
22	Soldani C and Scovassi AI: Poly(ADP-ribose) polymerase-1 cleavage during apoptosis: An update. Apoptosis. 7:321–328. 2002. View Article : Google Scholar : PubMed/NCBI
23	Murphy CS, Meisner LF, Wu SQ and Jordan VC: Short- and long-term estrogen deprivation of T47D human breast cancer cells in culture. Eur J Cancer Clin Oncol. 25:1777–1788. 1989. View Article : Google Scholar : PubMed/NCBI
24	Gray D, Jubb AM, Hogue D, Dowd P, Kljavin N, Yi S, Bai W, Frantz G, Zhang Z, Koeppen H, et al: Maternal embryonic leucine zipper kinase/murine protein serine-threonine kinase 38 is a promising therapeutic target for multiple cancers. Cancer Res. 65:9751–9761. 2005. View Article : Google Scholar : PubMed/NCBI
25	Davezac N, Baldin V, Blot J, Ducommun B and Tassan JP: Human pEg3 kinase associates with and phosphorylates CDC25B phosphatase: a potential role for pEg3 in cell cycle regulation. Oncogene. 21:7630–7641. 2002. View Article : Google Scholar : PubMed/NCBI
26	Mils V, Baldin V, Goubin F, Pinta I, Papin C, Waye M, Eychene A and Ducommun B: Specific interaction between 14-3-3 isoforms and the human CDC25B phosphatase. Oncogene. 19:1257–1265. 2000. View Article : Google Scholar : PubMed/NCBI
27	Forrest A and Gabrielli B: Cdc25B activity is regulated by 14-3-3. Oncogene. 20:4393–4401. 2001. View Article : Google Scholar : PubMed/NCBI
28	Seong HA, Gil M, Kim KT, Kim SJ and Ha H: Phosphorylation of a novel zinc-finger-like protein, ZPR9, by murine protein serine/threonine kinase 38 (MPK38). Biochem J. 361:597–604. 2002. View Article : Google Scholar : PubMed/NCBI
29	Seong HA, Kim KT and Ha H: Enhancement of B-MYB transcriptional activity by ZPR9, a novel zinc finger protein. J Biol Chem. 278:9655–9662. 2003. View Article : Google Scholar : PubMed/NCBI
30	Joaquin M and Watson RJ: Cell cycle regulation by the B-Myb transcription factor. Cell Mol Life Sci. 60:2389–2401. 2003. View Article : Google Scholar : PubMed/NCBI
31	Tao D, Pan Y, Lu H, Zheng S, Lin H, Fang H and Cao F: B-myb is a gene implicated in cell cycle and proliferation of breast cancer. Int J Clin Exp Pathol. 7:5819–5827. 2014.PubMed/NCBI
32	Zhang X, Tang N, Hadden TJ and Rishi AK: Akt, FoxO and regulation of apoptosis. Biochim Biophys Acta. 1813:1978–1986. 2011. View Article : Google Scholar : PubMed/NCBI
33	Hay N: Interplay between FOXO, TOR, and Akt. Biochim Biophys Acta. 1813:1965–1970. 2011. View Article : Google Scholar : PubMed/NCBI
34	Medema RH, Kops GJ, Bos JL and Burgering BM: AFX-like Forkhead transcription factors mediate cell-cycle regulation by Ras and PKB through p27kip1. Nature. 404:782–787. 2000. View Article : Google Scholar : PubMed/NCBI
35	Gu C, Banasavadi-Siddegowda YK, Joshi K, Nakamura Y, Kurt H, Gupta S and Nakano I: Tumor-specific activation of the C-JUN/MELK pathway regulates glioma stem cell growth in a p53-dependent manner. Stem Cells. 31:870–881. 2013. View Article : Google Scholar : PubMed/NCBI
36	Kim KH, Young BD and Bender JR: Endothelial estrogen receptor isoforms and cardiovascular disease. Mol Cell Endocrinol. 389:65–70. 2014. View Article : Google Scholar : PubMed/NCBI