Knockdown of SALL4 expression using RNA interference induces cell cycle arrest, enhances early apoptosis, inhibits invasion and increases chemosensitivity to temozolomide in U251 glioma cells

Zhang,Lei; Yan,Yong; Jiang,Ying; Qian,Jun; Jiang,Lei; Hu,Guohan; Lu,Yicheng; Luo,Chun

doi:10.3892/ol.2017.6722

Knockdown of SALL4 expression using RNA interference induces cell cycle arrest, enhances early apoptosis, inhibits invasion and increases chemosensitivity to temozolomide in U251 glioma cells

Authors:
- Lei Zhang
- Yong Yan
- Ying Jiang
- Jun Qian
- Lei Jiang
- Guohan Hu
- Yicheng Lu
- Chun Luo
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Affiliations: Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China
Published online on: August 4, 2017 https://doi.org/10.3892/ol.2017.6722
Pages: 4263-4269

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Abstract

Spalt-like transcription factor 4 (SALL4) is essential for the maintenance of the self‑renewal and pluripotent properties in embryonic stem cells. Although the detailed mechanism remains unclear, dysregulation of SALL4 has been detected in various malignancies. Previously, the authors' of the present study reported that the expression level of SALL4 was associated with the poor prognosis of glioblastoma multiforme (GBM). The present study aimed to investigate the function of SALL4 in U251 human glioblastoma cells, including apoptosis and invasion inhibition. It was revealed that knockdown of SALL4 expression through RNA interference induced cell cycle arrest, enhanced early apoptosis and significantly inhibited invasion. Furthermore, downregulation of SALL4 was associated with a significantly lower expression level of the core transcription factors, including POU class 5 homeobox 1, SRY‑box 2 and Nanog homeobox. In addition, inhibition of SALL4 significantly reduced the concentration of chemotherapeutic agent temozolomide required to inhibit cell growth by 50%, which decreased from 113.66±23.07 and 114.93±20.91 µg/ml to 68.34±3.52 and 67.44±4.71 µg/ml in two independent short interfering RNA transfected groups. These results indicate that SALL4 serves an important role in the GBM pathophysiology and targeting SALL4 may be a potential approach to the treatment of GBM.

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1	Weller M: Novel diagnostic and therapeutic approaches to malignant glioma. Swiss Med Wkly. 141:w132102011.PubMed/NCBI
2	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
3	Wen P and Kesari S: Malignant gliomas in adults. N Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI
4	Kohlhase J, Schuh R, Dowe G, Kühnlein RP, Jäckle H, Schroeder B, Schulz-Schaeffer W, Kretzschmar HA, Köhler A, Müller U, et al: Isolation, characterization, and organ-specific expression of two novel human zinc finger genes related to the Drosophila gene spalt. Genomics. 38:291–298. 1996. View Article : Google Scholar : PubMed/NCBI
5	Zhang J, Tam WL, Tong GQ, Wu Q, Chan HY, Soh BS, Lou Y, Yang J, Ma Y, Chai L, et al: Sall4 modulates embryonic stem cell pluripotency and early embryonic development by the transcriptional regulation of Pou5f1. Nat Cell Biol. 8:1114–1123. 2006. View Article : Google Scholar : PubMed/NCBI
6	Yang J, Gao C, Chai L and Ma Y: A novel SALL4/OCT4 transcriptional feedback network for pluripotency of embryonic stem cells. PLoS One. 5:e107662010. View Article : Google Scholar : PubMed/NCBI
7	Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J and Dirks PB: Identification of a cancer stem cell in human brain tumors. Cancer Res. 63:5821–5828. 2003.PubMed/NCBI
8	Galli R, Binda E, Orfanelli U, Cipelletti B, Gritti A, De Vitis S, Fiocco R, Foroni C, Dimeco F and Vescovi A: Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res. 64:7011–7021. 2004. View Article : Google Scholar : PubMed/NCBI
9	Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ and Clarke MF: Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA. 100:3983–3988. 2003. View Article : Google Scholar : PubMed/NCBI
10	Kobayashi D, Kuribayashi K, Tanaka M and Watanabe N: Overexpression of SALL4 in lung cancer and its importance in cell proliferation. Oncol Rep. 26:965–970. 2011.PubMed/NCBI
11	Kobayashi D, Kuribayshi K, Tanaka M and Watanabe N: SALL4 is essential for cancer cell proliferation and is overexpressed at early clinical stages in breast cancer. Int J Oncol. 38:933–939. 2011.PubMed/NCBI
12	Yong KJ, Gao C, Lim JS, Yan B, Yang H, Dimitrov T, Kawasaki A, Ong CW, Wong KF, Lee S, et al: Oncofetal gene SALL4 in aggressive hepatocellular carcinoma. N Engl J Med. 368:2266–2276. 2013. View Article : Google Scholar : PubMed/NCBI
13	Aguila JR, Liao W, Yang J, Avila C, Hagag N, Senzel L and Ma Y: SALL4 is a robust stimulator for the expansion of hematopoietic stem cells. Blood. 118:576–585. 2011. View Article : Google Scholar : PubMed/NCBI
14	Yang J, Aguila JR, Alipio Z, Lai R, Fink LM and Ma Y: Enhanced self-renewal of hematopoietic stem/progenitor cells mediated by the stem cell gene Sall4. J Hematol Oncol. 4:382011. View Article : Google Scholar : PubMed/NCBI
15	Zhang L, Yan Y, Jiang Y, Cui Y, Zou Y, Qian J, Luo C, Lu Y and Wu X: The expression of SALL4 in patients with gliomas: High level of SALL4 expression is correlated with poor outcome. J Neurooncol. 121:261–268. 2015. View Article : Google Scholar : PubMed/NCBI
16	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
17	Ma Y, Cui W, Yang J, Qu J, Di C, Amin HM, Lai R, Ritz J, Krause DS and Chai L: SALL4, a novel oncogene, is constitutively expressed in human acute myeloid leukemia (AML) and induces AML in transgenic mice. Blood. 108:2726–2735. 2006. View Article : Google Scholar : PubMed/NCBI
18	Cheng L, Alexander R, Zhang S, Pan CX, MacLennan GT, Lopez-Beltran A and Montironi R: The clinical and therapeutic implications of cancer stem cell biology. Expert Rev Anticancer Ther. 11:1131–1143. 2011. View Article : Google Scholar : PubMed/NCBI
19	Yu H, Lee H, Herrmann A, Buettner R and Jove R: Revisiting STAT3 signalling in cancer: New and unexpected biological functions. Nat Rev Cancer. 14:736–746. 2014. View Article : Google Scholar : PubMed/NCBI
20	Boyer LA, Lee TI, Cole MF, Johnstone SE, Levine SS, Zucker JP, Guenther MG, Kumar RM, Murray HL, Jenner RG, et al: Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 122:947–956. 2005. View Article : Google Scholar : PubMed/NCBI
21	Guo Y, Liu S, Wang P, Zhao S, Wang F, Bing L, Zhang Y, Ling EA, Gao J and Hao A: Expression of embryonic stem cell-associated genes Oct4, Sox2 and Nanog in human gliomas. Histopathology. 59:763–775. 2011. View Article : Google Scholar : PubMed/NCBI
22	Holmberg J, He X, Peredo I, Orrego A, Hesselager G, Ericsson C, Hovatta O, Oba-Shinjo SM, Marie SK, Nistér M and Muhr J: Activation of neural and pluripotent stem cell signatures correlates with increased malignancy in human glioma. PLoS One. 6:e184542011. View Article : Google Scholar : PubMed/NCBI
23	Shen DW, Pouliot LM, Hall MD and Gottesman MM: Cisplatin resistance: A cellular self-defense mechanism resulting from multiple epigenetic and genetic changes. Pharmacol Rev. 64:706–721. 2012. View Article : Google Scholar : PubMed/NCBI
24	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
25	Hiddingh L, Raktoe RS, Jeuken J, Hulleman E, Noske DP, Kaspers GJ, Vandertop WP, Wesseling P and Wurdinger T: Identification of temozolomide resistance factors in glioblastoma via integrative miRNA/mRNA regulatory network analysis. Sci Rep. 4:52602014. View Article : Google Scholar : PubMed/NCBI
26	Kaina B, Christmann M, Naumann S and Roos WP: MGMT: Key node in the battle against genotoxicity, carcinogenicity and apoptosis induced by alkylating agents. DNA Repair (Amst). 6:1079–1099. 2007. View Article : Google Scholar : PubMed/NCBI
27	Zeng SS, Yamashita T, Kondo M, Nio K, Hayashi T, Hara Y, Nomura Y, Yoshida M, Hayashi T, Oishi N, et al: The transcription factor SALL4 regulates stemness of EpCAM-positive hepatocellular carcinoma. J Hepatol. 60:127–134. 2014. View Article : Google Scholar : PubMed/NCBI
28	Kitange GJ, Mladek AC, Carlson BL, Schroeder MA, Pokorny JL, Cen L, Decker PA, Wu W, Lomberk GA, Gupta SK, et al: Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts. Clin Cancer Res. 18:4070–4079. 2012. View Article : Google Scholar : PubMed/NCBI
29	Kim HS, Kim NC, Chae KH, Kim G, Park WS, Park YK and Kim YW: Expression of multidrug resistance-associated protein 2 in human gallbladder carcinoma. Biomed Res Int. 2013:5275342013. View Article : Google Scholar : PubMed/NCBI
30	Chamberlain MC, Bota DA, Linskey ME and Schwartz PH: Neural stem/progenitors and glioma stem-like cells have differential sensitivity to chemotherapy. Neurology. 77:e135–e136. 2011. View Article : Google Scholar : PubMed/NCBI