Inhibition of GSK3 and MEK induced cancer stem cell generation via the Wnt and MEK signaling pathways

Liao,Shengtao; Gan,Li; Qin,Wanxiang; Liu,Chang; Mei,Zhechuan

doi:10.3892/or.2018.6600

Inhibition of GSK3 and MEK induced cancer stem cell generation via the Wnt and MEK signaling pathways

Authors:
- Shengtao Liao
- Li Gan
- Wanxiang Qin
- Chang Liu
- Zhechuan Mei
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Affiliations: Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, P.R. China, Teaching and Research Section of Forensic Medicine, College of Basic Medicine, Chongqing Medical University, Chongqing 400016, P.R. China, Department of Pain Management, Southwest Hospital, The First Affiliated Hospital of The Third Military Medical University, Chongqing 400038, P.R. China, Institute of Medicine and Hygienic Equipment for High Altitude Region, College of High Altitude Military Medicine, Third Military Medical University, Chongqing 400038, P.R. China
Published online on: July 25, 2018 https://doi.org/10.3892/or.2018.6600
Pages: 2005-2013
Copyright: © Liao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cancer stem cells (CSCs) are considered to be tumor‑initiating cells, responsible for tumor invasive growth and dissemination to distant organ sites. Typically, radiation treatment and chemotherapy should target CSCs. However, current research investigating CSCs is impeded by the difficulty of isolating pure CSCs and maintaining them in vitro. In the present study, the synergistic inhibition of glycogen synthase kinase 3 and mitogen‑activated protein kinase kinase using small molecules, CHIR99021 and PD184352, efficiently generated CSCs from immortalized human mammary epithelial cells (HMLEs) and resulted in the acquisition of mesenchymal traits and the expression of epithelial‑mesenchymal transition markers. The cell proliferation, invasion and migration of HMLE cells were significantly promoted by CHIR99021 and PD184352 (P<0.05). Furthermore, the cell cycle was shifted from the G0/G1 phase to the G2/M phase, and the apoptotic rate was suppressed in HMLE cells following treatment with CHIR99021 and PD184352. Compared with control group, the stimulated cells exhibited an increased ability to form mammospheres and regenerate a tumor. In addition to these properties, the induced cells also exhibited notable chemotherapy resistance. In vivo, the treatment of cells with CHIR99021 and PD184352 promoted the growth of HMLE‑engrafted tumor types. These results provide a practical strategy for the generation of CSCs using small molecules in vitro, which provides a cell resource that may be used for drug screening. Additionally, the present results additionally highlighted the synergistic functions of Wnt and mitogen‑activated protein kinase kinase signaling pathways in tumorigenesis.

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1	Reya T, Morrison SJ, Clarke MF and Weissman IL: Stem cells, cancer, and cancer stem cells. Nature. 414:105–111. 2001. View Article : Google Scholar : PubMed/NCBI
2	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: Accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
3	Dalerba P, Cho RW and Clarke MF: Cancer stem cells: Models and concepts. Annu Rev Med. 58:267–284. 2007. View Article : Google Scholar : PubMed/NCBI
4	Rowehl RA, Crawford H, Dufour A, Ju J and Botchkina GI: Genomic analysis of prostate cancer stem cells isolated from a highly metastatic cell line. Cancer Genomics Proteomics. 5:301–310. 2008.PubMed/NCBI
5	Abraham BK, Justenhoven C, Pesch B, Harth V, Weirich G, Baisch C, Rabstein S, Ko YD, Brüning T, Fischer HP, et al: Investigation of genetic variants of genes of the hemochromatosis pathway and their role in breast cancer. Cancer Epidemiol Biomarkers Prev. 14:1102–1107. 2005. View Article : Google Scholar : PubMed/NCBI
6	Hill RP and Perris R: ‘Destemming’ cancer stem cells. J Natl Cancer Inst. 99:1435–1440. 2007. View Article : Google Scholar : PubMed/NCBI
7	Zhu QS, Rosenblatt K, Huang KL, Lahat G, Brobey R, Bolshakov S, Nguyen T, Ding Z, Belousov R, Bill K, et al: Vimentin is a novel AKT1 target mediating motility and invasion. Oncogene. 30:457–470. 2011. View Article : Google Scholar : PubMed/NCBI
8	An WF, Germain AR, Bishop JA, Nag PP, Metkar S, Ketterman J, Walk M, Weiwer M, Liu X, Patnaik D, et al: Discovery of potent and highly selective inhibitors of GSK3b. 2010.
9	Reya T and Clevers H: Wnt signalling in stem cells and cancer. Nature. 434:843–850. 2005. View Article : Google Scholar : PubMed/NCBI
10	Pellicano F, Simara P, Sinclair A, Helgason GV, Copland M, Grant S and Holyoake TL: The MEK inhibitor PD184352 enhances BMS-214662-induced apoptosis in CD34⁺ CML stem/progenitor cells. Leukemia. 25:1159–1167. 2011. View Article : Google Scholar : PubMed/NCBI
11	Li L, Bennett SA and Wang L: Role of E-cadherin and other cell adhesion molecules in survival and differentiation of human pluripotent stem cells. Cell Adh Migr. 6:59–70. 2012. View Article : Google Scholar : PubMed/NCBI
12	Satelli A and Li S: Vimentin in cancer and its potential as a molecular target for cancer therapy. Cell Mol Life Sci. 68:3033–3046. 2011. View Article : Google Scholar : PubMed/NCBI
13	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
14	Kawano K, Efferson CL, Peoples GE, Carter D, Tsuda N, Murray JL and Ioannides CG: Sensitivity of undifferentiated, high-TCR density CD8⁺ cells to methylene groups appended to tumor antigen determines their differentiation or death. Cancer Res. 65:2930–2937. 2005. View Article : Google Scholar : PubMed/NCBI
15	Kruger NJ: The Bradford method for protein quantitationMethods in Molecular Biology: Basic Protein and Peptide Potocols. Chapter 32. Walker JM: Humana Press; Totowa, NJ: pp. 9–15. 1994
16	Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA and Lander ES: Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell. 138:645–659. 2009. View Article : Google Scholar : PubMed/NCBI
17	Vinogradov S and Wei X: Cancer stem cells and drug resistance: The potential of nanomedicine. Nanomedicine (Lond). 7:597–615. 2012. View Article : Google Scholar : PubMed/NCBI
18	Facompre N, Nakagawa H, Herlyn M and Basu D: Stem-like cells and therapy resistance in squamous cell carcinomas. Adv Pharmacol. 65:235–265. 2012. View Article : Google Scholar : PubMed/NCBI
19	Huang SD, Yuan Y, Tang H, Liu XH, Fu CG, Cheng HZ, Bi JW, Yu YW, Gong DJ, Zhang W, et al: Tumor cells positive and negative for the common cancer stem cell markers are capable of initiating tumor growth and generating both progenies. PLoS One. 8:e545792013. View Article : Google Scholar : PubMed/NCBI
20	Jaggupilli A and Elkord E: Significance of CD44 and CD24 as cancer stem cell markers: An enduring ambiguity. Clin Dev Immunol. 2012:7080362012. View Article : Google Scholar : PubMed/NCBI
21	Li Y, Wang L, Pappan L, Galliher-Beckley A and Shi J: IL-1^β promotes stemness and invasiveness of colon cancer cells through Zeb1 activation. Mol Cancer. 11:872012. View Article : Google Scholar : PubMed/NCBI
22	Wang ZL, Fan ZQ, Jiang HD and Qu JM: Selective Cox-2 inhibitor celecoxib induces epithelial-mesenchymal transition in human lung cancer cells via activating MEK-ERK signaling. Carcinogenesis. 34:638–646. 2013. View Article : Google Scholar : PubMed/NCBI
23	Xie G, Yao Q, Liu Y, Du S, Liu A, Guo Z, Sun A, Ruan J, Chen L, Ye C and Yuan Y: IL-6-induced epithelial-mesenchymal transition promotes the generation of breast cancer stem-like cells analogous to mammosphere cultures. Int J Oncol. 40:1171–1179. 2012.PubMed/NCBI
24	Hur EM and Zhou FQ: GSK3 signalling in neural development. Nat Rev Neurosci. 11:539–551. 2010. View Article : Google Scholar : PubMed/NCBI
25	Fisar Z and Hroudová J: Intracellular signalling pathways and mood disorders. Folia Biol (Praha). 56:135–148. 2010.PubMed/NCBI
26	Zou Y, Liu FY, Wu J, Wan L, Fang SF, Zhang ZY, Luo Y, Chen MH, Huang MZ, He M and Huang OP: Mutational analysis of the RAS/RAF/MEK/ERK signaling pathway in 260 Han Chinese patients with cervical carcinoma. Oncol Lett. 14:2427–2431. 2017. View Article : Google Scholar : PubMed/NCBI
27	Xiong Y, Huang F, Li X, Chen Z, Feng D, Jiang H, Chen W and Zhang X: CCL21/CCR7 interaction promotes cellular migration and invasion via modulation of the MEK/ERK1/2 signaling pathway and correlates with lymphatic metastatic spread and poor prognosis in urinary bladder cancer. Int J Oncol. 51:75–90. 2017. View Article : Google Scholar : PubMed/NCBI
28	Santarpia L, Lippman SM and El-Naggar AK: Targeting the MAPK-RAS-RAF signaling pathway in cancer therapy. Expert Opin Ther Targets. 16:103–119. 2012. View Article : Google Scholar : PubMed/NCBI
29	Singh AM, Reynolds D, Cliff T, Ohtsuka S, Mattheyses AL, Sun Y, Menendez L, Kulik M and Dalton S: Signaling network crosstalk in human pluripotent cells: A Smad2/3-regulated switch that controls the balance between self-renewal and differentiation. Cell Stem Cell. 10:312–326. 2012. View Article : Google Scholar : PubMed/NCBI
30	Gong K, Zhou F, Huang H, Gong Y and Zhang L: Suppression of GSK3^β by ERK mediates lipopolysaccharide induced cell migration in macrophage through ^β-catenin signaling. Protein Cell. 3:762–768. 2012. View Article : Google Scholar : PubMed/NCBI
31	Sears R, Nuckolls F, Haura E, Taya Y, Tamai K and Nevins JR: Multiple Ras-dependent phosphorylation pathways regulate Myc protein stability. Genes Dev. 14:2501–2514. 2000. View Article : Google Scholar : PubMed/NCBI
32	Dave B, Mittal V, Tan NM and Chang JC: Epithelial-mesenchymal transition, cancer stem cells and treatment resistance. Breast Cancer Res. 14:2022012. View Article : Google Scholar : PubMed/NCBI
33	Wang Y and Zhou BP: Epithelial-mesenchymal transition in breast cancer progression and metastasis. Chin J Cancer. 30:603–611. 2011. View Article : Google Scholar : PubMed/NCBI
34	Xiang J, Fu X, Ran W and Wang Z: Grhl2 reduces invasion and migration through inhibition of TGFβ-induced EMT in gastric cancer. Oncogenesis. 6:e2842017. View Article : Google Scholar : PubMed/NCBI
35	Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, et al: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 133:704–715. 2008. View Article : Google Scholar : PubMed/NCBI
36	Phillips TM, McBride WH and Pajonk F: The response of CD24(−/low)/CD44⁺ breast cancer-initiating cells to radiation. J Natl Cancer Inst. 98:1777–1785. 2006. View Article : Google Scholar : PubMed/NCBI
37	Andarawewa KL, Erickson AC, Chou WS, Costes SV, Gascard P, Mott JD, Bissell MJ and Barcellos-Hoff MH: Ionizing radiation predisposes nonmalignant human mammary epithelial cells to undergo transforming growth factor beta induced epithelial to mesenchymal transition. Cancer Res. 67:8662–8670. 2007. View Article : Google Scholar : PubMed/NCBI
38	Sineva GS and Pospelov VA: Inhibition of GSK3beta enhances both adhesive and signalling activities of beta-catenin in mouse embryonic stem cells. Biol Cell. 102:549–564. 2012. View Article : Google Scholar