Effect of temozolomide on livin and caspase-3 in U251 glioma stem cells

Li,Genhua; Zhang,Hao; Liu,Yang; Kong,Lingsheng; Guo,Qiang; Jin,Feng

doi:10.3892/etm.2014.2144

March-2015 Volume 9 Issue 3

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March-2015 Volume 9 Issue 3

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Article Open Access

Effect of temozolomide on livin and caspase-3 in U251 glioma stem cells

Authors:
- Genhua Li
- Hao Zhang
- Yang Liu
- Lingsheng Kong
- Qiang Guo
- Feng Jin
View Affiliations / Copyright

Affiliations: Neuro-oncology Laboratory, Department of Neurosurgery, Affiliated Hospital of Jining Medical College, Jining, Shandong 272029, P.R. China

Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 744-750
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Published online on: December 18, 2014

https://doi.org/10.3892/etm.2014.2144
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Abstract

The aim of the present study was to analyze the effect of temozolomide (TMZ) on the antiapoptotic gene livin and the associated gene caspase-3. Cancer stem cells were isolated from U251 glioblastoma cells using immunomagnetic beads. The glioma cells and glioma stem cells were transfected with livin or small hairpin RNA (shRNA) against livin using lentiviral vectors. Quantitative PCR, flow cytometry and a Cell Counting kit‑8 assay were used to detect the expression of livin and caspase-3, analyze the cell cycle and investigate cell proliferation, respectively, following treatment with various concentrations of TMZ (0, 25, 50, 100, 200 and 400 µmol/l) for different periods of time (24, 48 and 72 h). The expression levels of livin and caspase‑3 in the U251 stem cells were significantly higher than those in the U251 cells (P<0.01). At the same intervention time, the expression levels of livin decreased and those of caspase-3 increased as the concentration of TMZ increased (P<0.05). The expression levels of livin and caspase-3 in the U251 cells were lower than those in the U251 stem cells with the same intervention time and concentration of TMZ (P<0.05). The cell cycle was arrested in the G2/M phase in the U251 cells following TMZ intervention; the proportion of cells in the G2/M phase increased as the concentration of TMZ increased (P<0.05). The U251 stem cells were arrested in the S phase following treatment with TMZ; the proportion of cells in the S phase increased as the concentration of TMZ increased (P<0.05). In conclusion, the expression levels of livin and caspase-3 were effectively inhibited and increased, respectively, in all cell models following treatment with TMZ. TMZ is able to arrest the cell cycle and enhance cell apoptosis. U251 stem cells are less vulnerable than U251 cells to TMZ.

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1	Hingtgen S, Ren XH, Terwilliger E, et al: Targeting multiple pathways in glioma with stem cell and viral delivered S-TRAIL and temozolomide. Mol Cancer Ther. 7:3575–3585. 2008. View Article : Google Scholar : PubMed/NCBI
2	Norden AD and Wen PY: Glioma therapy in adults. Neurologist. 12:279–292. 2006. View Article : Google Scholar : PubMed/NCBI
3	Liu GT, Yuan XP, Zeng ZH, et al: Analysis of gene expression and chemoresistance of CD133⁺ cancer stem cells in glioblastoma. Mol Cancer. 5:67–79. 2006. View Article : Google Scholar
4	Dean M, Fojo T and Bates S: Tumour stem cells and drug resistance. Nat Rev Cancer. 5:275–284. 2005. View Article : Google Scholar : PubMed/NCBI
5	Gilbert CA and Ross AH: Cancer stem cell: cell culture, markers and targets for new therapies. J Cell Biochem. 108:1031–1038. 2009. View Article : Google Scholar : PubMed/NCBI
6	Beier D, Röhrl S, Pillai DR, et al: Temozolomide preferentially depletes cancer stem cells in glioblastoma. Cancer Res. 68:5706–5715. 2008. View Article : Google Scholar : PubMed/NCBI
7	Jin F, Zhao L, Zhao HY, et al: Comparison between cells and cancer stem-like cells isolated from glioblastoma and astrocytoma on expression of anti-apoptotic and multidrug resistance-associated protein genes. Neuroscience. 154:541–550. 2008. View Article : Google Scholar : PubMed/NCBI
8	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
9	LaCasse EC, Baird S, Korneluk RG and MacKenzie AE: The inhibitors of apoptosis (IAPs) and their emerging role in cancer. Oncogene. 17:3247–3259. 1998. View Article : Google Scholar
10	Lopes RB, Gangeswaran R, McNeish IA, et al: Expression of the IAP protein family is dysregulated in pancreatic cancer cells and is important for resistance to chemotherapy. Int J Cancer. 120:2344–2352. 2007. View Article : Google Scholar : PubMed/NCBI
11	Yan B: Research progress on livin protein: an inhibitor of apoptosis. Mol Cell Biochem. 357:39–45. 2011. View Article : Google Scholar : PubMed/NCBI
12	Nachmias B, Ashab Y, Bucholtz V, et al: Caspase-mediated cleavage converts livin from an antiapoptotic to a proapoptotic factor: implications for drug-resistant melanoma. Cancer Res. 63:6340–6349. 2003.PubMed/NCBI
13	Jin F, Zhao L, Zhao HY, et al: Paradoxical expression of anti-apoptotic and MRP genes on cancer stem-like cell isolated for TJ905 glioblastoma multiforme cell line. Cancer Invest. 26:338–343. 2008. View Article : Google Scholar : PubMed/NCBI
14	Jin F, Zhao L, Guo YJ, et al: Influence of Etoposide on antiapoptotic and multidrug resistance-associated protein genes in CD133 positive U251 glioblastoma stem-like cells. Brain Res. 1336:103–111. 2010. View Article : Google Scholar : PubMed/NCBI
15	Ma YB and Chang HY: Caspase work model during pathogen infection. Virol Sin. 26:366–375. 2011. View Article : Google Scholar : PubMed/NCBI
16	He X, Nie H, Hong Y, et al: SIRT2 activity is required for the survival of C6 glioma cells. Biochem Biophys Res Commun. 417:468–472. 2012. View Article : Google Scholar
17	Ryu CH, Yoon WS, Park KY, et al: Valproic acid downregulates the expression of MGMT and sensitizes temozolomide-resistant glioma cells. J Biomed Biotechnol. 1–9. 2012. View Article : Google Scholar
18	Fisher T, Galanti G, Lavie G, et al: Mechanisms operative in the antitumor activity of temozolomide in glioblastoma multiforme. Cancer J. 13:335–344. 2007. View Article : Google Scholar : PubMed/NCBI
19	Beier D, Schulz JB and Beier CP: Chemoresistance of glioblastoma cancer stem cells - much more complex than expected. Mol Cancer. 10:128–139. 2011. View Article : Google Scholar :
20	Overmeyer JH, Young AM, Bhanot H and Maltese WA: A chalcone-related small molecule that induces methuosis, a novel form of non-apoptotic cell death in glioblastoma cells. Mol Cancer. 10:69–86. 2011. View Article : Google Scholar

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Effect of temozolomide on livin and caspase-3 in U251 glioma stem cells

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