Inhibition of the Rac1-WAVE2-Arp2/3 signaling pathway promotes radiosensitivity via downregulation of cofilin-1 in U251 human glioma cells

Zhou,Tao; Wang,Chen‑Han; Yan,Hua; Zhang,Rui; Zhao,Jin‑Bing; Qian,Chun‑Fa; Xiao,Hong; Liu,Hong‑Yi

doi:10.3892/mmr.2016.5088

Inhibition of the Rac1-WAVE2-Arp2/3 signaling pathway promotes radiosensitivity via downregulation of cofilin-1 in U251 human glioma cells

Authors:
- Tao Zhou
- Chen‑Han Wang
- Hua Yan
- Rui Zhang
- Jin‑Bing Zhao
- Chun‑Fa Qian
- Hong Xiao
- Hong‑Yi Liu
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Affiliations: Department of Neurosurgery, Nanjing Medical University, Affiliated Nanjing Brain Hospital, Nanjing, Jiangsu 210029, P.R. China, Neuropsychiatric Institute, Nanjing Medical University, Affiliated Nanjing Brain Hospital, Nanjing, Jiangsu 210029, P.R. China
Published online on: April 5, 2016 https://doi.org/10.3892/mmr.2016.5088
Pages: 4414-4420

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Abstract

The Ras-related C3 botulinum toxin substrate 1 (Rac1)-WASP-family verprolin-homologous protein-2 (WAVE2)-actin-related protein 2/3 (Arp2/3) signaling pathway has been identified to be involved in cell migration and invasion in various types of cancer cell. Cofilin‑1 (CFL‑1), which is regulated by the Rac1‑WAVE2‑Arp2/3 signaling pathway, may promote radioresistance in glioma. Therefore, the present study aimed to investigate the potential role of the Rac1‑WAVE2‑Arp2/3 signaling pathway in radioresistance in U251 human glioma cells and elucidate its affect on CFL‑1 expression. Western blot analysis was performed to evaluate the protein expression of CFL‑1. In the present study, Rac1 was inhibited by NSC 23766, WAVE2 was inhibited by transfection with short hairpin (sh)RNA‑WAVE2 using Lipofectamine™ 2000 and Arp2/3 was inhibited by CK‑666. Cell viability was measured using the 3‑(4,5‑dimethylthiazol‑2‑yl)-2,5‑diphenyltetrazolium bromide assay, the cell migration ability was examined by a wound‑healing assay, and the cell invasion ability was assessed using a Transwell culture chamber system. The results showed that inhibition of the Rac1‑WAVE2‑Arp2/3 signaling pathway using NSC 23766, shRNA‑WAVE2 or CK‑666 reduced the cell viability, migration and invasion abilities in U251 human glioma cells, concordant with a reduced expression of CFL‑1. Furthermore, the expression of CFL‑1 was significantly increased in radioresistant U251 glioma cells when compared with normal U251 human glioma cells. These findings indicate that inhibition of the Rac1‑WAVE2‑Arp2/3 signaling pathway may promote radiosensitivity, which may partially result from the downregulation of CFL‑1 in U251 human glioma cells.

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1	Ford E, Catt S, Chalmers A and Fallowfield L: Systematic review of supportive care needs in patients with primary malignant brain tumors. Neuro Oncol. 14:392–404. 2012. View Article : Google Scholar : PubMed/NCBI
2	Pace A, Metro G and Fabi A: Supportive care in neurooncology. Curr Opin Oncol. 22:621–626. 2010. View Article : Google Scholar : PubMed/NCBI
3	Catt S, Chalmers A and Fallowfield L: Psychosocial and supportive-care needs in high-grade glioma. Lancet Oncol. 9:884–891. 2008. View Article : Google Scholar : PubMed/NCBI
4	Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, et al: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 352:987–996. 2005. View Article : Google Scholar : PubMed/NCBI
5	Shapiro JR, Yung WK and Shapiro WR: Isolation, karyotype and clonal growth of heterogeneous subpopulations of human malignant gliomas. Cancer Res. 41:2349–2359. 1981.PubMed/NCBI
6	Bonavia R, Inda MM, Cavenee WK and Furnari FB: Heterogeneity maintenance in glioblastoma: A social network. Cancer Res. 71:4055–4060. 2011. View Article : Google Scholar : PubMed/NCBI
7	Sottoriva A, Spiteri I, Piccirillo SG, Touloumis A, Collins VP, Marioni JC, Curtis C, Watts C and Tavaré S: Intratumor heterogeneity in human glioblastoma reflects cancer evolutionary dynamics. Proc Natl Acad Sci USA. 110:4009–4014. 2013. View Article : Google Scholar : PubMed/NCBI
8	Walker MD, Strike TA and Sheline GE: An analysis of dose-effect relationship in the radiotherapy of malignant gliomas. Int J Radiat Oncol Biol Phys. 5:1725–1731. 1979. View Article : Google Scholar : PubMed/NCBI
9	Huang L, Li B, Tang S, Guo H, Li W, Huang X, Yan W and Zou F: Mitochondrial KATP channels control glioma radioresistance by regulating ROS-Induced ERK activation. Mol Neurobiol. 52:626–637. 2015. View Article : Google Scholar
10	Michaelson D, Abidi W, Guardavaccaro D, Zhou M, Ahearn I, Pagano M and Philips MR: Rac1 accumulates in the nucleus during the G2 phase of the cell cycle and promotes cell division. J Cell Biol. 181:485–496. 2008. View Article : Google Scholar : PubMed/NCBI
11	Prudnikova TY, Rawat SJ and Chernoff J: Molecular pathways: Targeting the kinase effectors of RHO-family GTPases. Clin Cancer Res. 21:24–29. 2015. View Article : Google Scholar :
12	Skvortsov S, Dudás J, Eichberger P, Witsch-Baumgartner M, Loeffler-Ragg J, Pritz C, Schartinger VH, Maier H, Hall J, Debbage P, et al: Rac1 as a potential therapeutic target for chemo-radioresistant head and neck squamous cell carcinomas (HNSCC). Br J Cancer. 110:2677–2687. 2014. View Article : Google Scholar : PubMed/NCBI
13	Machesky LM and Insall RH: Signaling to actin dynamics. J Cell Biol. 146:267–272. 1999. View Article : Google Scholar : PubMed/NCBI
14	Pollard TD and Borisy GG: Cellular motility driven by assembly and disassembly of actin filaments. Cell. 112:453–465. 2003. View Article : Google Scholar : PubMed/NCBI
15	Lane J, Martin T, Weeks HP and Jiang WG: Structure and role of WASP and WAVE in Rho GTPase signalling in cancer. Cancer Genomics Proteomics. 11:155–165. 2014.PubMed/NCBI
16	Ma X, Espana-Serrano L, Kim WJ, Thayele PH, Nie Z and Daaka Y: βArrestin1 regulates the guanine nucleotide exchange factor RasGRF2 expression and the small GTPase Rac-mediated formation of membrane protrusion and cell motility. J Biol Chem. 289:13638–13650. 2014. View Article : Google Scholar : PubMed/NCBI
17	Koestler SA, Steffen A, Nemethova M, Winterhoff M, Luo N, Holleboom JM, Krupp J, Jacob S, Vinzenz M, Schur F, et al: Arp2/3 complex is essential for actin network treadmilling as well as for targeting of capping protein and cofilin. Mol Biol Cell. 24:2861–2875. 2013. View Article : Google Scholar : PubMed/NCBI
18	Ko HS, Kim JS, Cho SM, Lee HJ, Ahn KS, Kim SH and Lee EO: Urokinase-type plasminogen activator expression and Rac1/WAVE-2/Arp2/3 pathway are blocked by pterostilbene to suppress cell migration and invasion in MDA-MB-231 cells. Bioorg Med Chem Lett. 24:1176–1179. 2014. View Article : Google Scholar : PubMed/NCBI
19	Bamburg JR: Proteins of the ADF/cofilin family: Essential regulators of actin dynamics. Annu Rev Cell Dev Biol. 15:185–230. 1999. View Article : Google Scholar : PubMed/NCBI
20	Carlier MF, Ressad F and Pantaloni D: Control of actin dynamics in cell motility. Role of ADF/cofilin. J Biol Chem. 274:33827–33830. 1999. View Article : Google Scholar : PubMed/NCBI
21	Maciver SK and Hussey PJ: The ADF/cofilin family: Actin-remodeling proteins. Genome Biol. 3:reviews30072002. View Article : Google Scholar : PubMed/NCBI
22	Moriyama K and Yahara I: The actin-severing activity of cofilin is exerted by the interplay of three distinct sites on cofilin and essential for cell viability. Biochem J. 365:147–155. 2002. View Article : Google Scholar : PubMed/NCBI
23	Gurniak CB, Perlas E and Witke W: The actin depolymerizing factor n-cofilin is essential for neural tube morphogenesis and neural crest cell migration. Dev Biol. 278:231–241. 2005. View Article : Google Scholar : PubMed/NCBI
24	Yan H, Yang K, Xiao H, Zou YJ, Zhang WB and Liu HY: Over-expression of cofilin-1 and phosphoglycerate kinase 1 in astrocytomas involved in pathogenesis of radioresistance. Cns Neurosci Ther. 18:729–736. 2012. View Article : Google Scholar : PubMed/NCBI
25	Du HQ, Chen L, Wang Y, Wang LJ, Yan H, Liu HY and Xiao H: Increasing radiosensitivity with the downregulation of cofilin-1 in U251 human glioma cells. Mol Med Rep. 11:3354–3360. 2015.
26	Dang I, Gorelik R, Sousa-Blin C, Derivery E, Guérin C, Linkner J, Nemethova M, Dumortier JG, Giger FA, Chipysheva TA, et al: Inhibitory signalling to the Arp2/3 complex steers cell migration. Nature. 503:281–284. 2013.PubMed/NCBI
27	Ho JN, Kang GY, Lee SS, Kim J, Bae IH, Hwang SG and Um HD: Bcl-XL and STAT3 mediate malignant actions of gamma-irradiation in lung cancer cells. Cancer Sci. 101:1417–1423. 2010. View Article : Google Scholar : PubMed/NCBI
28	Cheng JC, Chou CH, Kuo ML and Hsieh CY: Radiation-enhanced hepatocellular carcinoma cell invasion with MMP-9 expression through PI3K/Akt/NF-kappaB signal transduction pathway. Oncogene. 25:7009–7018. 2006. View Article : Google Scholar : PubMed/NCBI
29	Park CM, Park MJ, Kwak HJ, Lee HC, Kim MS, Lee SH, Park IC, Rhee CH and Hong SI: Ionizing radiation enhances matrix metalloproteinase-2 secretion and invasion of glioma cells through Src/epidermal growth factor receptor-mediated p38/Akt and phosphatidylinositol 3-kinase/Akt signaling pathways. Cancer Res. 66:8511–8519. 2006. View Article : Google Scholar : PubMed/NCBI
30	Wang W, Eddy R and Condeelis J: The cofilin pathway in breast cancer invasion and metastasis. Nat Rev Cancer. 7:429–440. 2007. View Article : Google Scholar : PubMed/NCBI
31	Le Clainche C, Schlaepfer D, Ferrari A, Klingauf M, Grohmanova K, Veligodskiy A, Didry D, Le D, Egile C, Carlier MF and Kroschewski R: IQGAP1 stimulates actin assembly through the N-WASP-Arp2/3 pathway. J Biol Chem. 282:426–435. 2007. View Article : Google Scholar
32	DesMarais V, Macaluso F, Condeelis J and Bailly M: Synergistic interaction between the Arp2/3 complex and cofilin drives stimulated lamellipod extension. J Cell Sci. 117:3499–3510. 2004. View Article : Google Scholar : PubMed/NCBI
33	Arnold CR, Abdelmoez A, Thurner G, Debbage P, Lukas P, Skvortsov S and Skvortsova II: Rac1 as a multifunctional therapeutic target to prevent and combat cancer metastasis. Oncoscience. 1:513–521. 2014. View Article : Google Scholar
34	Gupta GP and Massagué J: Cancer metastasis: Building a framework. Cell. 127:679–695. 2006. View Article : Google Scholar : PubMed/NCBI
35	Wang C, Lu S, Jiang J, Jia X, Dong X and Bu P: Hsa-microRNA-101 suppresses migration and invasion by targeting Rac1 in thyroid cancer cells. Oncol Lett. 8:1815–1821. 2014.PubMed/NCBI
36	Lorenz M, Yamaguchi H, Wang Y, Singer RH and Condeelis J: Imaging sites of N-wasp activity in lamellipodia and invadopodia of carcinoma cells. Curr Biol. 14:697–703. 2004. View Article : Google Scholar : PubMed/NCBI
37	Bernstein BW and Bamburg JR: ADF/cofilin: A functional node in cell biology. Trends Cell Biol. 20:187–195. 2010. View Article : Google Scholar : PubMed/NCBI
38	Sreekumar R, Sayan BS, Mirnezami AH and Sayan AE: MicroRNA control of invasion and metastasis pathways. Front Genet. 2:582011. View Article : Google Scholar