Abstract
The new claim about the origin of cancer known as Cancer Stem Cell theory states that a somatic differentiated cell can dedifferentiated or reprogrammed for regaining the cancer cell features. It has been recently shown that expression of stemness factors such as Oct4, Sox2, Nanog and Klf4, in a variety of somatic cancers can leads to development of tumorogenesis. Here, the expression of Oct4 variants were evaluated in brain tumor tissues by quantitative RT-PCR and immunohistochemical (IHC) analysis. In next phase of our study, the expression of Oct4B1 was knock-down in brain cancer cell lines and its effect on cell cycle was assessed. Finally, in order to get insights into sequence-structure-function relationships of Oct4 isofroms, their sequences were analysed using bioinformatic tools. Our data revealed that all three variants of Oct4 are expressed in different types of brain cancer. The expression level of Oct4B1, in contast to Oct4B, was much higher in high-grade brain tumors compared with low-grade ones. In line with qPCR, the expression of Oct4A and B isofroms was confirmed with IHC in different types of brain tumors. Moreover, as a result of the suppression of Oct4B1 expression, the brain cancer cells were arrested in G2/M phase of cell cycle. Bioinfromatics data indicated that the predicted Oct4B1 protein have DNA binding properties. All together, our findings suggest that Oct4B1 has a potential role in tumorigenesis of brain cancer and can be considered as a new tumor marker with potential value in diagnosis and treatment of brain cancer.
Similar content being viewed by others
References
Al-Hajj M, Clarke MF (2004) Self-renewal and solid tumor stem cells. Oncogene 23:7274–7282
Clarke MF, Fuller M (2006) Stem cells and cancer: two faces of eve. Cell 124:1111–1115
Cao L, Li C, Shen S, Yan Y, Ji W, Wang J, Qian H, Jiang X, Li Z, Wu M (2013) OCT4 increases BIRC5 and CCND1 expression and promotes cancer progression in hepatocellular carcinoma. BMC Cancer 13:82
Wen J, Park JY, Park KH, Chung HW, Bang S, Park SW, Song SY (2010) Oct4 and Nanog expression is associated with early stages of pancreatic carcinogenesis. Pancreas 39:622–626
Asadi MH, Derakhshani A, Mowla SJ (2014) Concomitant upregulation of nucleostemin and downregulation of Sox2 and Klf4 in gastric adenocarcinoma. Tumour Biol 35:7177–7185
Rothenberg ME, Clarke MF, Diehn M (2010) The Myc connection: ES cells and cancer. Cell 143:184–186
Clarissa NA, Brad AB (2015) Enrichment of the embryonic stem cell reprogramming factors Oct4, Nanog, Myc, and Sox2 in benign and malignant vascular tumors. BMC Clin Pathol 15:18
Radzisheuskaya A, Silva JC (2014) Do all roads lead to Oct4? the emerging concepts of induced pluripotency. Trends Cell Biol 24:275–284
Tsai CC, Su PF, Huang YF, Yew TL, Hung SC (2012) Oct4 and Nanog directly regulate Dnmt1 to maintain self-renewal and undifferentiated state in mesenchymal stem cells. Mol Cell 47:169–182
Bouillez A, Rajabi H, Pitroda S, Jin C, Alam M, Kharbanda A, Tagde A, Wong KK, Kufe D (2016) Inhibition of MUC1-C suppresses MYC expression and attenuates malignant growth in KRAS mutant lung adenocarcinomas. Cancer Res 76:1538–1548
Tagde A, Rajabi H, Bouillez A, Alam M, Gali R, Bailey S, Tai YT, Hideshima T, Anderson K, Avigan D, Kufe D (2016) MUC1-C drives MYC in multiple myeloma. Blood 127:2587–2597
Rajabi H, Tagde A, Alam M, Bouillez A, Pitroda S, Suzuki Y, Kufe D (2016) DNA methylation by DNMT1 and DNMT3b methyltransferases is driven by the MUC1-C oncoprotein in human carcinoma cells. Oncogene. doi:10.1038/onc.2016.180
Tagde A, Rajabi H, Stroopinsky D, Gali R, Alam M, Bouillez A, Kharbanda S, Stone R, Avigan D, Kufe D (2016) MUC1-C induces DNA methyltransferase 1 and represses tumor suppressor genes in acute myeloid leukemia. Oncotarget 7:38974–38987
Nath S, Mukherjee P (2014) MUC1: a multifaceted oncoprotein with a key role in cancer progression. Trends Mol Med 20:332–342
Takahashi H, Jin C, Rajabi H, Pitroda S, Alam M, Ahmad R, Raina D, Hasegawa M, Suzuki Y, Tagde A, Bronson RT, Weichselbaum R, Kufe D (2015) MUC1-C activates the TAK1 inflammatory pathway in colon cancer. Oncogene 34:5187–5197
Hasegawa M, Takahashi H, Rajabi H, Alam M, Suzuki Y, Yin L, Tagde A, Maeda T, Hiraki M, Sukhatme VP, Kufe D (2016) Functional interactions of the cystine/glutamate antiporter, CD44v and MUC1-C oncoprotein in triple-negative breast cancer cells. Oncotarget 7:11756–11769
Lopez-Bertoni H, Lal B, Li A, Caplan M, Guerrero-Cázares H, Eberhart CG, Quiñones-Hinojosa A, Glas M, Scheffler B, Laterra J, Li Y (2015) DNMT-dependent suppression of microRNA regulates the induction of GBM tumor-propagating phenotype by Oct4 and Sox2. Oncogene 34:3994–4004
Takeda J, Seino S, Bell GI (1992) Human Oct3 gene family: cDNA sequences, alternative splicing, gene organization, chromosomal location, and expression at low levels in adult tissues. Nucleic Acids Res 20:4613–4620
Atlasi Y, Mowla SJ, Ziaee SA, Gokhale PJ, Andrews PW (2008) OCT4 spliced variants are differentially expressed in human pluripotent and nonpluripotent cells. Stem Cells 26:3068–3074
Asadi MH, Mowla SJ, Fathi F, Aleyasin A, Asadzadeh J, Atlasi Y (2011) OCT4B1, a novel spliced variant of OCT4, is highly expressed in gastric cancer and acts as an antiapoptotic factor. Int J Cancer 128:2645–2652
Asadzadeh J, Asadi MH, Shakhssalim N, Rafiee MR, Kalhor HR, Tavallaei M, Mowla SJ (2012) A plausible anti-apoptotic role of up-regulated OCT4B1 in bladder tumors. Urol J 9:574–580
Mirzaei MR, Najafi A, Arababadi MK, Asadi MH, Mowla SJ (2014) Altered expression of apoptotic genes in response to OCT4B1 suppression in human tumor cell lines. Tumour Biol 35:9999–10009
Mirzaei MR, Kazemi Arababadi M, Asadi MH, Mowla SJ (2016) Altered expression of high molecular weight heat shock proteins after OCT4B1 suppression in human tumor cell lines. Cell J 17:608–616
Mirzaei MR, Asadi MH, Mowla SJ, Hassanshahi G, Ahmadi Z (2016) Down-regulation of HSP40 gene family following OCT4B1 suppression in human tumor cell lines. Iran J Basic Med Sci 19:187–193
Farashahi Yazd E, Rafiee MR, Soleimani M, Tavallaei M, Salmani MK, Mowla SJ (2011) OCT4B1, a novel spliced variant of OCT4, generates a stable truncated protein with a potential role in stress response. Cancer Lett 309:170–175
Sonawane P, Cho HE, Tagde A, Verlekar D, Yu AL, Reynolds CP, Kang MH (2014) Metabolic characteristics of 13-cis-retinoic acid (isotretinoin) and anti-tumour activity of the 13-cis-retinoic acid metabolite 4-oxo-13-cis-retinoic acid in neuroblastoma. Br J Pharmacol 171:5330–5344
Tagde A, Singh H, Kang MH, Reynolds CP (2014) The glutathione synthesis inhibitor buthionine sulfoximine synergistically enhanced melphalan activity against preclinical models of multiple myeloma. Blood Cancer J 4:e229
Rice P, Longden I, Bleasby A.EMBOSS (2000) The European molecular biology open software suite. Trends Genet 16:276–277
Goujon M, McWilliam H, Li W, Valentin F, Squizzato S, Paern J, Lopez R (2010) A new bioinformatics analysis tools framework at EMBL-EBI. Nucleic Acids Res 38:695–699
McWilliam H, Li W, Uludag M, Squizzato S, Park YM, Buso N, Cowley AP, Lopez R (2013) Analysis tool web services from the EMBL-EBI. Nucleic Acids Res 41:597–600
Sievers F, Wilm A, Dineen D, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Söding J, Thompson JD, Higgins DG (2011) Fast, scalable generation of high-quality protein multiple sequence alignments using clustal omega. Mol Syst Biol 7:539
Marchler-Bauer A, Zheng C, Chitsaz F, Derbyshire MK, Geer LY, Geer RC et al (2013) CDD: conserved domains and protein three-dimensional structure. Nucleic Acids Res 41:348–352
Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T et al (2014) SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res 42:252–258
Arnold K, Bordoli L, Kopp J, Schwede T (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22:195–201
Gasteiger E, Hoogland C, Gattiker A, Duvaud S, Wilkins MR, Appel RD, Bairoch A (2005) Protein identification and analysis tools on the ExPASy server. Proteomics protocols handbook. Humana Press, New York
Esch D, Vahokoski J, Groves MR, Pogenberg V, Cojocaru V, Vom Bruch H et al (2013) A unique Oct4 interface is crucial for reprogramming to pluripotency. Nat Cell Biol 15:295–301
Acknowledgments
The study was supported by Kerman Graduate University of Advanced Technology (grant No.1/2536). All biological materials were provided by the IRAN NATIONAL TUMOR BANK which is funded by the Cancer Institute of Tehran University, for Cancer Research.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Asadi, M.H., Khalifeh, K. & Mowla, S.J. OCT4 spliced variants are highly expressed in brain cancer tissues and inhibition of OCT4B1 causes G2/M arrest in brain cancer cells. J Neurooncol 130, 455–463 (2016). https://doi.org/10.1007/s11060-016-2255-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11060-016-2255-1