All-trans retinoic acid influences viability, migration and adhesion of U251 glioblastoma cells

Jelena Marjanović Vićentić, Marija Schwirtlich, Nataša Kovačević-Grujičić, Milena Stevanović, Danijela Drakulić


Glioblastoma (GBM) is one of the most aggressive and deadly forms of cancer. Literature data reveals that all-trans retinoic acid (ATRA) has anticancer effects on different types of tumor cells. However, data about the effects of ATRA on glioblastoma cells are contradictory. In this study, we examined whether ATRA treatment affects features of human glioblastoma U251 cells. To that end, the cells were treated with different concentrations of ATRA. Results obtained by MTT and the crystal violet assays imply that ATRA affected the viability of U251 glioblastoma cells in a dose- and time-dependent manner. Fluorescence staining of microtubule cytoskeleton protein α-tubulin revealed that ATRA induced changes in cell morphology. Using semi-quantitative RT-PCR we found that the expression of SOX3 and GFAP genes, as markers of neural differentiation, was not changed upon ATRA treatment. Thus, the observed changes in cell morphology after ATRA treatment are not associated with neural differentiation of U251 glioblastoma cells. The scratch-wound healing assay revealed that ATRA changed the mode of U251 cell migration from collective to single cell motility. The cell-matrix adhesion assay demonstrated that the pharmacologically relevant concentration of ATRA lowered the cell-matrix adhesion capability of U251 cells. In conclusion, our results imply that further studies are needed before ATRA could be considered for the treatment of glioblastoma.

Received: March 27, 2017; Revised: May 23, 2017; Accepted: June 2, 2017; Published online: June 12, 2017

How to cite this article: Marjanović Vićentić J, Schwirtlich M, Kovačević-Grujičić N, Stevanović M, Drakulić D. All-trans retinoic acid influences viability, migration and adhesion of U251 glioblastoma cells. Arch Biol Sci. 2017;69(4):699-706.


glioblastoma; ATRA; differentiation; viability; cell migration

Full Text:



Adamski V, Schmitt AD, Fluh C, Synowitz M, Hattermann K, Held-Feindt J. Isolation and characterization of fast migrating human glioma cells in the progression of malignant gliomas. Oncol Res. 2017;25(3):341-53.

Rao SS, Lannutti JJ, Viapiano MS, Sarkar A, Winter JO. Toward 3D biomimetic models to understand the behavior of glioblastoma multiforme cells. Tissue Eng Part B Rev. 2014;20:314-27.

Sottoriva A, Spiteri I, Piccirillo SG, Touloumis A, Collins VP, Marioni JC, Curtis C, Watts C, Tavare S. Intratumor heterogeneity in human glioblastoma reflects cancer evolutionary dynamics. Proc Natl Acad Sci U S A. 2013;110:4009-14.

Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, Curschmann J, Janzer RC, Ludwin SK, Gorlia T, Allgeier A, Lacombe D, Cairncross JG, Eisenhauer E, Mirimanoff RO, European Organisation for R, Treatment of Cancer Brain T, Radiotherapy G, National Cancer Institute of Canada Clinical Trials G. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987-96.

Castellino RC, Durden DL. Mechanisms of disease: the PI3K-Akt-PTEN signaling node--an intercept point for the control of angiogenesis in brain tumors. Nat Clin Pract Neurol. 2007;3:682-93.

Fathima Hurmath K, Ramaswamy P, Nandakumar DN. IL-1beta microenvironment promotes proliferation, migration, and invasion of human glioma cells. Cell Biol Int. 2014;38:1415-22.

Haque A, Das A, Hajiaghamohseni LM, Younger A, Banik NL, Ray SK. Induction of apoptosis and immune response by all-trans retinoic acid plus interferon-gamma in human malignant glioblastoma T98G and U87MG cells. Cancer Immunol Immunother. 2007;56:615-25.

Kagechika H, Shudo K. Synthetic retinoids: recent developments concerning structure and clinical utility. J Med Chem. 2005;48:5875-83.

Liang C, Yang L, Guo S. All-trans retinoic acid inhibits migration, invasion and proliferation, and promotes apoptosis in glioma cells in vitro. Oncol Lett. 2015;9:2833-8.

Xia SL, Wu ML, Li H, Wang JH, Chen NN, Chen XY, Kong QY, Sun Z, Liu J. CRABP-II- and FABP5-independent responsiveness of human glioblastoma cells to all-trans retinoic acid. Oncotarget. 2015;6:5889-902.

Fenaux P. The role of all-trans-retinoic acid in the treatment of acute promyelocytic leukemia. Acta Haematol. 1993;89(Suppl 1):22-7.

Simeone AM, Tari AM. How retinoids regulate breast cancer cell proliferation and apoptosis. Cell Mol Life Sci. 2004;61:1475-84.

Pasquali D, Chieffi P, Deery WJ, Nicoletti G, Bellastella A, Sinisi AA. Differential effects of all-trans-retinoic acid (RA) on Erk1/2 phosphorylation and cAMP accumulation in normal and malignant human prostate epithelial cells: Erk1/2 inhibition restores RA-induced decrease of cell growth in malignant prostate cells. Eur J Endocrinol. 2005;152:663-9.

Zheng Y, Kramer PM, Lubet RA, Steele VE, Kelloff GJ, Pereira MA. Effect of retinoids on AOM-induced colon cancer in rats: modulation of cell proliferation, apoptosis and aberrant crypt foci. Carcinogenesis. 1999;20:255-60.

Herreros-Villanueva M, Er TK, Bujanda L. Retinoic Acid Reduces Stem Cell-Like Features in Pancreatic Cancer Cells. Pancreas. 2015;44:918-24.

Schug TT, Berry DC, Shaw NS, Travis SN, Noy N. Opposing effects of retinoic acid on cell growth result from alternate activation of two different nuclear receptors. Cell. 2007;129:723-33.

Bouterfa H, Picht T, Kess D, Herbold C, Noll E, Black PM, Roosen K, Tonn JC. Retinoids inhibit human glioma cell proliferation and migration in primary cell cultures but not in established cell lines. Neurosurgery. 2000;46:419-30.

Schug TT, Berry DC, Toshkov IA, Cheng L, Nikitin AY, Noy N. Overcoming retinoic acid-resistance of mammary carcinomas by diverting retinoic acid from PPARbeta/delta to RAR. Proc Natl Acad Sci U S A. 2008;105:7546-51.

Tang K, Cao L, Fan SQ, Wu MH, Huang H, Zhou YH, Zhou M, Tang YL, Wang R, Zeng F, Liao P, Li XL, Li GY. [Effect of all-trans-retinoic acid on C6 glioma cell proliferation and differentiation]. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2008;33:892-7.

Lee HJ, Park IH, Kim HJ, Kim SU. Human neural stem cells overexpressing glial cell line-derived neurotrophic factor in experimental cerebral hemorrhage. Gene Ther. 2009;16:1066-76.

Drakulic D, Krstic A, Stevanovic M. Establishment and initial characterization of SOX2-overexpressing NT2/D1 cell clones. Genet Mol Res. 2012;11:1385-400.

Yan M, Liu Q. Differentiation therapy: a promising strategy for cancer treatment. Chin J Cancer. 2016;35:3.

Lu J, Zhang F, Zhao D, Hong L, Min J, Zhang L, Li F, Yan Y, Li H, Ma Y, Li Q. ATRA-inhibited proliferation in glioma cells is associated with subcellular redistribution of beta-catenin via up-regulation of Axin. J Neurooncol. 2008;87:271-7.

Torsvik A, Stieber D, Enger PO, Golebiewska A, Molven A, Svendsen A, Westermark B, Niclou SP, Olsen TK, Chekenya Enger M, Bjerkvig R. U-251 revisited: genetic drift and phenotypic consequences of long-term cultures of glioblastoma cells. Cancer Med. 2014;3:812-24.

Pijuan-Thompson V, Grammer JR, Stewart J, Silverstein RL, Pearce SF, Tuszynski GP, Murphy-Ullrich JE, Gladson CL. Retinoic acid alters the mechanism of attachment of malignant astrocytoma and neuroblastoma cells to thrombospondin-1. Exp Cell Res. 1999;249:86-101.

Papi A, Bartolini G, Ammar K, Guerra F, Ferreri AM, Rocchi P, Orlandi M. Inhibitory effects of retinoic acid and IIF on growth, migration and invasiveness in the U87MG human glioblastoma cell line. Oncol Rep. 2007;18:1015-21.

Paillaud E, Costa S, Fages C, Plassat JL, Rochette-Egly C, Monville C, Tardy M. Retinoic acid increases proliferation rate of GL-15 glioma cells, involving activation of STAT-3 transcription factor. J Neurosci Res. 2002;67:670-9.

Das A, Banik NL, Ray SK. Retinoids induced astrocytic differentiation with down regulation of telomerase activity and enhanced sensitivity to taxol for apoptosis in human glioblastoma T98G and U87MG cells. J Neurooncol. 2008;87:9-22.

Ying M, Wang S, Sang Y, Sun P, Lal B, Goodwin CR, Guerrero-Cazares H, Quinones-Hinojosa A, Laterra J, Xia S. Regulation of glioblastoma stem cells by retinoic acid: role for Notch pathway inhibition. Oncogene. 2011;30:3454-67.

Zeng Y, Yang Z, Xu JG, Yang MS, Zeng ZX, You C. Differentially expressed genes from the glioblastoma cell line SHG-44 treated with all-trans retinoic acid in vitro. J Clin Neurosci. 2009;16:285-94.

Shi Z, Lou M, Zhao Y, Zhang Q, Cui D, Wang K. Effect of all-trans retinoic acid on the differentiation of U87 glioma stem/progenitor cells. Cell Mol Neurobiol. 2013;33:943-51.

Liotta LA. Tumor invasion and metastases--role of the extracellular matrix: Rhoads Memorial Award lecture. Cancer Res. 1986;46:1-7.

Sternlicht MD, Werb Z. How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol. 2001;17:463-516.

Brosicke N, Faissner A. Role of tenascins in the ECM of gliomas. Cell Adh Migr. 2015;9:131-40.

Toda D, Ota T, Tsukuda K, Watanabe K, Fujiyama T, Murakami M, Naito M, Shimizu N. Gefitinib decreases the synthesis of matrix metalloproteinase and the adhesion to extracellular matrix proteins of colon cancer cells. Anticancer Res. 2006;26:129-34.

Todd JR, Ryall KA, Vyse S, Wong JP, Natrajan RC, Yuan Y, Tan AC, Huang PH. Systematic analysis of tumour cell-extracellular matrix adhesion identifies independent prognostic factors in breast cancer. Oncotarget. 2016;7:62939-53.

Cavallaro U, Christofori G. Cell adhesion in tumor invasion and metastasis: loss of the glue is not enough. Biochim Biophys Acta. 2001;1552:39-45.

Jiang WG, Sanders AJ, Katoh M, Ungefroren H, Gieseler F, Prince M, Thompson SK, Zollo M, Spano D, Dhawan P, Sliva D, Subbarayan PR, Sarkar M, Honoki K, Fujii H, Georgakilas AG, Amedei A, Niccolai E, Amin A, Ashraf SS, Ye L, Helferich WG, Yang X, Boosani CS, Guha G, Ciriolo MR, Aquilano K, Chen S, Azmi AS, Keith WN, Bilsland A, Bhakta D, Halicka D, Nowsheen S, Pantano F, Santini D. Tissue invasion and metastasis: Molecular, biological and clinical perspectives. Semin Cancer Biol. 2015;35(Suppl):S244-S275.

Chattopadhyay N, Ray S, Biswas N, Chatterjee A. Effect of all-trans-retinoic acid on integrin receptors of human cervical cancer (SiHa) cells. Gynecol Oncol. 1999;75:215-21.

Li Y, Francia G, Zhang JY. p62/IMP2 stimulates cell migration and reduces cell adhesion in breast cancer. Oncotarget. 2015;6(32):32656-68.

Park JE, Tan HS, Datta A, Lai RC, Zhang H, Meng W, Lim SK, Sze SK. Hypoxic tumor cell modulates its microenvironment to enhance angiogenic and metastatic potential by secretion of proteins and exosomes. Mol Cell Proteomics. 2010;9:1085-99.

Hendrix MJ, Wood WR, Seftor EA, Lotan D, Nakajima M, Misiorowski RL, Seftor RE, Stetler-Stevenson WG, Bevacqua SJ, Liotta LA, Sobel ME, Raz A, Lotan R. Retinoic acid inhibition of human melanoma cell invasion through a reconstituted basement membrane and its relation to decreases in the expression of proteolytic enzymes and motility factor receptor. Cancer Res. 1990;50:4121-30.


  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.