Association between SET expression and glioblastoma cell apoptosis and proliferation

He,Kunyan; Shi,Lihong; Jiang,Tingting; Li,Qiang; Chen,Yao; Meng,Chuan

doi:10.3892/ol.2016.4951

Association between SET expression and glioblastoma cell apoptosis and proliferation

Authors:
- Kunyan He
- Lihong Shi
- Tingting Jiang
- Qiang Li
- Yao Chen
- Chuan Meng
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Affiliations: Department of Anatomy, Basic Medical and Forensic Medical Institute, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Department of Neurosurgery, The Third People's Hospital of Chengdu, Chengdu, Sichuan 610041, P.R. China
Published online on: August 3, 2016 https://doi.org/10.3892/ol.2016.4951
Pages: 2435-2444
Copyright: © He et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Glioblastoma multiforme (GBM) was one of the first cancer types systematically studied at a genomic and transcriptomic level due to its high incidence and aggressivity; however, the detailed mechanism remains unclear, even though it is known that numerous cytokines are involved in the occurrence and development of GBM. The present study aimed to determine whether the SET gene has a role in human glioblastoma carcinogenesis. A total of 32 samples, including 18 cases of glioma, 2 cases of meningioma and 12 normal brain tissue samples, were detected using the streptavidin‑peroxidase method through immunohistochemistry. To reduce SET gene expression in U251 and U87MG cell lines, the RNA interference technique was used and transfection with small interfering (si)RNA of the SET gene was performed. Cell apoptosis was detected by flow cytometry, cell migration was examined by Transwell migration assay and cell proliferation was determined by Cell Counting Kit‑8. SET, Bcl‑2, Bax and caspase‑3 mRNA and protein expression levels were detected by reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. Positive protein expression of SET was observed in the cell nucleus, with the expression level of SET significantly higher in glioma tissues compared with normal brain tissue (P=0.001). Elevated expression of SET was significantly associated with gender (P=0.002), tumors classified as World Health Organization grade II (P=0.031), III (P=0.003) or IV (P=0.001), and moderately (P=0.031) or poorly differentiated (P=0.001) tumors. Compared with the negative and non‑treatment (blank) control cells, SET gene expression was significantly inhibited (P=0.006 and P<0.001), cell apoptosis was significantly increased (P=0.001 and P<0.001), cell proliferation was significantly inhibited (P=0.002 and P=0.015), and cell migration was significantly decreased (P=0.001 and P=0.001) in siRNA‑transfected U87MG‑SET and U251‑SET cells, respectively. In addition, mRNA and protein expression levels of Bcl‑2 were significantly inhibited in U87MG‑SET and U251‑SET cells, while mRNA and protein expression levels of Bax and caspase‑3 were significantly increased, compared with the two control groups. Thus, the current data suggests that SET may regulate the proliferation and apoptosis of glioblastoma cells by upregulating Bcl‑2, and downregulating Bax and caspase‑3.

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1	Wu CX, Lin GS, Lin ZX, Zhang JD, Liu SY and Zhou CF: Peritumoral edema shown by MRI predicts poor clinical outcome in glioblastoma. World J Surg Oncol. 13:972015. View Article : Google Scholar : PubMed/NCBI
2	Thakkar JP, Dolecek TA, Horbinski C, Ostrom QT, Lightner DD, BarnholtzSloan JS and Villano JL: Epidemiologic and molecular prognostic review of glioblastoma. Cancer Epidemiol Biomarkers Prev. 23:1985–1996. 2014. View Article : Google Scholar : PubMed/NCBI
3	Zhang L, Wang H, Ding K and Xu J: FTY720 induces autophagy-related apoptosis and necroptosis in human glioblastoma cells. Toxicol Lett. 236:43–59. 2015. View Article : Google Scholar : PubMed/NCBI
4	Koo S, Martin G and Toussaint LG: MicroRNA-145 promotes the phenotype of human glioblastoma cells selected for invasion. Anticancer Res. 35:3209–3215. 2015.PubMed/NCBI
5	Chakrabarti M and Ray SK: Direct transfection of miR-137 mimics is more effective than DNA demethylation of miR-137 promoter to augment anti-tumor mechanisms of delphinidin in human glioblastoma U87MG and LN18 cells. Gene. 573:141–152. 2015. View Article : Google Scholar : PubMed/NCBI
6	Wang G, Wang J, Zhao H, Wang J and Tony To SS: The role of Myc and let-7a in glioblastoma, glucose metabolism and response to therapy. Arch Biochem Biophys. 580:84–92. 2015. View Article : Google Scholar : PubMed/NCBI
7	Cimino PJ, Bredemeyer A, Abel HJ and Duncavage EJ: A wide spectrum of EGFR mutations in glioblastoma is detected by a single clinical oncology targeted next-generation sequencing panel. Exp Mol Pathol. 98:568–573. 2015. View Article : Google Scholar : PubMed/NCBI
8	Yan J, Kong LY, Hu J, Gabrusiewicz K, Dibra D, Xia X, Heimberger AB and Li S: FGL2 as a multimodality regulator of Tumor-mediated immune suppression and therapeutic target in gliomas. J Natl Cancer Inst. 107:pii.djv1372015. View Article : Google Scholar
9	Karetsou Z, Emmanouilidou A, Sanidas I, Liokatis S, Nikolakaki E, Politou AS and Papamarcaki T: Identification of distinct SET/TAF-Ibeta domains required for core histone binding and quantitative characterisation of the interaction. BMC Biochem. 10:102009. View Article : Google Scholar : PubMed/NCBI
10	Irie A, Harada K, Araki N and Nishimura Y: Phosphorylation of SET protein at Ser171 by protein kinase D2 diminishes its inhibitory effect on protein phosphatase 2A. PLoS One. 7:e512422012. View Article : Google Scholar : PubMed/NCBI
11	Li M, Makkinje A and Damuni Z: The myeloid leukemia-associated protein SET Is a potent inhibitor of protein phosphatase 2A. J Biol Chem. 271:11059–11062. 1996. View Article : Google Scholar : PubMed/NCBI
12	Almeida LO, Garcia CB, MatosSilva FA, Curti C and Leopoldino AM: Accumulated SET protein up-regulates and interacts with hnRNPK, increasing its binding to nucleic acids, the Bcl-xS repression, and cellular proliferation. Biochem Biophys Res Commun. 445:196–202. 2014. View Article : Google Scholar : PubMed/NCBI
13	Xu Z, Yang W, Shi N, Gao Y, Teng M and Niu L: Cloning, purification, crystallization and preliminary X-ray crystallographic analysis of SET/TAF-Iß δN from Homo sapiens. Acta Crystallogr Sect F Struct Biol Cryst Commun. 66:926–928. 2010. View Article : Google Scholar : PubMed/NCBI
14	Lam BD, Anthony EC and Hordijk PL: Cytoplasmic targeting of the proto-oncogene SET promotes cell spreading and migration. FEBS Lett. 587:111–119. 2013. View Article : Google Scholar : PubMed/NCBI
15	Cristóbal I, Garcia-Orti L, Cirauqui C, Cortes-Lavaud X, García-Sánchez MA, Calasanz MJ and Odero MD: Overexpression of SET is a recurrent event associated with poor outcome and contributes to protein phosphatase 2A inhibition in acute myeloid leukemia. Haematologica. 97:543–550. 2012. View Article : Google Scholar : PubMed/NCBI
16	Mukhopadhyay A, Tabanor K, Chaguturu R and Aldrich JV: Targeting inhibitor 2 of protein phosphatase 2A as a therapeutic strategy for prostate cancer treatment. Cancer Biol Ther. 14:962–972. 2013. View Article : Google Scholar : PubMed/NCBI
17	Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW and Kleihues P: The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol. 114:97–109. 2007. View Article : Google Scholar : PubMed/NCBI
18	Gonzales MF: Grading of gliomas. J Clin Neurosci. 4:16–18. 1997. View Article : Google Scholar : PubMed/NCBI
19	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
20	Kontos CK, Fendri A, Khabir A, MokdadGargouri R and Scorilas A: Quantitative expression analysis and prognostic significance of the BCL2-associated X gene in nasopharyngeal carcinoma: A retrospective cohort study. BMC Cancer. 13:2932013. View Article : Google Scholar : PubMed/NCBI
21	Manoochehri M, Karbasi A, Bandehpour M and Kazemi B: Down-regulation of BAX gene during carcinogenesis and acquisition of resistance to 5-FU in colorectal cancer. Pathol Oncol Res. 20:301–307. 2014. View Article : Google Scholar : PubMed/NCBI
22	Croci DO, Cogno IS, Vittar NB, Salvatierra E, Trajtenberg F, Podhajcer OL, Osinaga E, Rabinovich GA and Rivarola VA: Silencing survivin gene expression promotes apoptosis of human breast cancer cells through a caspase-independent pathway. J Cell Biochem. 105:381–390. 2008. View Article : Google Scholar : PubMed/NCBI
23	Choi JE, Kang SH, Lee SJ and Bae YK: Prognostic significance of Bcl-2 expression in non-basal triple-negative breast cancer patients treated with anthracycline-based chemotherapy. Tumour Biol. 35:12255–12263. 2014. View Article : Google Scholar : PubMed/NCBI
24	Gómez-Crisóstomo NP, López-Marure R, Zapata E, Zazueta C and Martinez-Abundis E: Bax induces cytochrome c release by multiple mechanisms in mitochondria from MCF7 cells. J Bioenerg Biomembr. 45:441–448. 2013. View Article : Google Scholar : PubMed/NCBI
25	Kavanagh E, Rodhe J, Burguillos MA, Venero JL and Joseph B: Regulation of caspase-3 processing by cIAP2 controls the switch between pro-inflammatory activation and cell death in microglia. Cell Death Dis. 5:e15652014. View Article : Google Scholar : PubMed/NCBI
26	Liu X, He Y, Li F, Huang Q, Kato TA, Hall RP and Li CY: caspase-3 promotes genetic instability and carcinogenesis. Mol Cell. 58:284–296. 2015. View Article : Google Scholar : PubMed/NCBI
27	Amar SK, Goyal S, Mujtaba SF, Dwivedi A, Kushwaha HN, Verma A, Chopra D, Chaturvedi RK and Ray RS: Role of type I & type II reactions in DNA damage and activation of caspase 3 via mitochondrial pathway induced by photosensitized benzophenone. Toxicol Lett. 235:84–95. 2015. View Article : Google Scholar : PubMed/NCBI
28	Wen X, Zhu J, Dong L and Chen Y: The role of c2orf68 and PI3K/Akt/mTOR pathway in human colorectal cancer. Med Oncol. 31:922014. View Article : Google Scholar : PubMed/NCBI