Activating transcription factor 3 is overexpressed in human glioma and its knockdown in glioblastoma cells causes growth inhibition both in vitro and in vivo

Ma,Siqi; Pang,Changhe; Song,Laijun; Guo,Fuyou; Sun,Hongwei

doi:10.3892/ijmm.2015.2173

Activating transcription factor 3 is overexpressed in human glioma and its knockdown in glioblastoma cells causes growth inhibition both in vitro and in vivo

Authors:
- Siqi Ma
- Changhe Pang
- Laijun Song
- Fuyou Guo
- Hongwei Sun
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Affiliations: Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China
Published online on: April 7, 2015 https://doi.org/10.3892/ijmm.2015.2173
Pages: 1561-1573
Copyright: © Ma et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Glioblastomas are highly malignant gliomas that are extremely invasive with high rates of recurrence and mortality. It has been reported that activating transcription factor 3 (ATF3) is expressed in elevated levels in multiple malignant tumors. The purpose of this study was to investigate the function of ATF3 in the development of glioma and its clinical significance. Immunohistochemical staining, western blot analysis and RT-qPCR revealed that the mRNA and protein levels of ATF3 and matrix metalloproteinase 2 (MMP2) were higher in the glioma than in the normal human brain tissues, and that their levels were proportional to the pathological grades. By contrast, the mRNA and protein levels of mammary serine protease inhibitor (maspin; SERPINB5) were significantly lower in the glioma than in the normal brain tissue, and maspin expression was inversely proportional to the glioma pathological grade. The transfection of U373MG glioblastoma cells with ATF3-siRNA induced a number of changes in cell behavior; the cell proliferative activity was decreased and flow cytometry revealed an increased proportion of cells arrested in the G0/G1 phase of the cell cycle. In addition, TUNEL staining indicated an increased proportion of cells undergoing apoptosis and Transwell assays revealed impaired cell mobility. The sizes of the tumors grown as xenografts in nude mice were also significantly reduced by treatment of host mice with ATF3-siRNA. Taken together, these results suggest that ATF3 promotes the progression of human gliomas.

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1	Pelzer AE, Bektic J, Haag P, Berger AP, Pycha A, Schäfer G, Rogatsch H, Horninger W, Bartsch G and Klocker H: The expression of transcription factor activating transcription factor 3 in the human prostate and its regulation by androgen in prostate cancer. J Urol. 175:1517–1522. 2006. View Article : Google Scholar : PubMed/NCBI
2	Janz M, Hummel M, Truss M, et al: Classical Hodgkin lymphoma is characterized by high constitutive expression of activating transcription factor 3 (ATF3), which promotes viability of Hodgkin/Reed-Sternberg cells. Blood. 107:2536–2539. 2006. View Article : Google Scholar
3	Iyengar P, Combs TP, Shah SJ, et al: Adipocyte-secreted factors synergistically promote mammary tumorigenesis through induction of anti-apoptotic transcriptional programs and protooncogene stabilization. Oncogene. 22:6408–6423. 2003. View Article : Google Scholar : PubMed/NCBI
4	Hai T and Curran T: Cross-family dimerization of transcription factors Fos/Jun and ATF/CREB alters DNA binding specificity. Proc Natl Acad Sci USA. 88:3720–3724. 1991. View Article : Google Scholar : PubMed/NCBI
5	Wolfgang CD, Chen BP, Martindale JL, Holbrook NJ and Hai T: gadd153/Chop10, a potential target gene of the transcriptional repressor ATF3. Mol Cell Biol. 17:6700–6707. 1997.PubMed/NCBI
6	Chen BPC, Liang G, Whelan J and Hai T: ATF3 and ATF3 delta Zip. Transcriptional repression versus activation by alternatively spliced isoforms. J Biol Chem. 269:15819–15826. 1994.PubMed/NCBI
7	Chen Z, Fan Z, McNeal JE, Nolley R, Caldwell MC, Mahadevappa M, Zhang Z, Warrington JA and Stamey TA: Hepsin and maspin are inversely expressed in laser capture microdissectioned prostate cancer. J Urol. 169:1316–1319. 2003. View Article : Google Scholar : PubMed/NCBI
8	Forsyth PA, Wong H, Laing TD, et al: Gelatinase-A (MMP-2), gelatinase-B (MMP-9) and membrane type matrix metallopro-teinase-1 (MT1-MMP) are involved in different aspects of the pathophysiology of malignant gliomas. Br J Cancer. 79:1828–1835. 1999. View Article : Google Scholar : PubMed/NCBI
9	Louis David N, Ohgaki Hiroko, Wiestler Otmar D, Cavenee Webster K, et al: Astrocytic tumours. Who Classification of Tumours of the Central Nervous System. World Health Organization; Geneva: pp. 13–45. 2007
10	Gatalica Z, Lele SM, Rampy BA and Norris BA: The expression of Fhit protein is related inversely to disease progression in patients with breast carcinoma. Cancer. 88:1378–1383. 2000. View Article : Google Scholar : PubMed/NCBI
11	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
12	Yin X, Dewille JW and Hai T: A potential dichotomous role of ATF3, an adaptive-response gene, in cancer development. Oncogene. 27:2118–2127. 2008. View Article : Google Scholar
13	Ishiguro T, Nakajima M, Naito M, Muto T and Tsuruo T: Identification of genes differentially expressed in B16 murine melanoma sublines with different metastatic potentials. Cancer Res. 56:875–879. 1996.PubMed/NCBI
14	Bandyopadhyay S, Wang Y, Zhan R, et al: The tumor metastasis suppressor gene Drg-1 down-regulates the expression of activating transcription factor 3 in prostate cancer. Cancer Res. 66:11983–11990. 2006. View Article : Google Scholar : PubMed/NCBI
15	Ishiguro T, Nagawa H, Naito M and Tsuruo T: Inhibitory effect of ATF3 antisense oligonucleotide on ectopic growth of HT29 human colon cancer cells. Jpn J Cancer Res. 91:833–836. 2000. View Article : Google Scholar : PubMed/NCBI
16	Lu D, Wolfgang CD and Hai T: Activating transcription factor 3, a stress-inducible gene, suppresses Rasstimulated tumorigenesis. J Biol Chem. 281:10473–10481. 2006. View Article : Google Scholar : PubMed/NCBI
17	Bottone FG Jr, Moon Y, Kim JS, Alston-Mills B, Ishibashi M and Eling TE: The anti-invasive activity of cyclooxygenase inhibitors is regulated by the transcription factor ATF3 (activating transcription factor 3). Mol Cancer Ther. 4:693–703. 2005. View Article : Google Scholar : PubMed/NCBI
18	Zhang W and Zhang M: Tissue microarray analysis of maspin expression and its reverse correlation with mutant p53 in various tumors. Int J Oncol. 20:1145–1150. 2002.PubMed/NCBI
19	Zou Z, Anisowicz A, Hendrix MJ, Thor A, Neveu M, Sheng S, Rafidi K, Seftor E and Sager R: Maspin, a serpin with tumor-suppressing activity in human mammary epithelial cells. Science. 263:526–529. 1994. View Article : Google Scholar : PubMed/NCBI
20	Wang DL, Wang YF, Shi GS and Huang H: Correlation of hTERT expression to maspin and bFGF expression and their significance in glioma. Ai Zheng. 26:601–606. 2007.In Chinese. PubMed/NCBI
21	Song H, Li Y, Lee J, Schwartz AL and Bu G: Low-density lipoprotein receptor-related protein 1 promotes cancer cell migration and invasion by inducing the expression of matrix metalloproteinases 2 and 9. Cancer Res. 69:879–886. 2009. View Article : Google Scholar : PubMed/NCBI
22	Kodera T, Nakagawa T, Kubota T, Kabuto M, Sato K and Kobayashi H: The expression and activation of matrix metallo-proteinase-2 in rat brain after implantation of C6 rat glioma cells. J Neurooncol. 46:105–114. 2000. View Article : Google Scholar
23	Musso O, Théret N, Campion JP, Turlin B, Milani S, Grappone C and Clément B: In situ detection of matrix metalloproteinase-2 (MMP2) and the metalloproteinase inhibitor TIMP2 transcripts in human primaryhepatocellular carcinoma and in liver metastasis. J Hepatol. 26:593–605. 1997. View Article : Google Scholar : PubMed/NCBI
24	Mendes O, Kim HT and Stoica G: Expression of MMP2, MMP9 and MMP3 in breast cancer brain metastasis in a rat model. Clin Exp Metastasis. 22:237–246. 2005. View Article : Google Scholar : PubMed/NCBI
25	Danilewicz M, Sikorska B and Wagrowska-Danilewicz M: Prognostic significance of the immunoexpression of matrix metalloproteinaseMMP2 and its inhibitor TIMP2 in laryngeal cancer. Med Sci Monit. 9:MT42–MT47. 2003.PubMed/NCBI
26	Herbst RS, Onn A and Sandler A: Angiogenesis and lung cancer: Prognostic and therapeutic implications. J Clin Oncol. 23:3243–3256. 2005. View Article : Google Scholar : PubMed/NCBI
27	Bodenstine TM, Seftor RE, Khalkhali-Ellis Z, Seftor EA, Pemberton PA and Hendrix MJ: Maspin: Molecular mechanisms and therapeutic implications. Cancer Metastasis Rev. 31:529–551. 2012. View Article : Google Scholar : PubMed/NCBI
28	Sheng S: The promise and challenge toward the clinical application of maspin in cancer. Front Biosci. 9:2733–2745. 2004. View Article : Google Scholar : PubMed/NCBI
29	Ngamkitidechakul C, Warejcka DJ, Burke JM, O’Brien WJ and Twining SS: Sufficiency of the reactive site loop of maspin for induction of cell-matrix adhesion and inhibition of cell invasion. Conversion of ovalbumin to a maspin-like molecule. J Biol Chem. 278:31796–31806. 2003. View Article : Google Scholar : PubMed/NCBI
30	Zou Z, Zhang W, Young D, Gleave MG, Rennie P, Connell T, Connelly R, Moul J, Srivastava S and Sesterhenn I: Maspin expression profile in human prostate cancer (CaP) and in vitro induction of Maspin expression by androgen ablation. Clin Cancer Res. 8:1172–1177. 2002.PubMed/NCBI
31	Hai T, Wolford CC and Chang YS: ATF3, a hub of the cellular adaptive-response network, in the pathogenesis of diseases: Is modulation of inflammation a unifying component? Gene Expr. 15:1–11. 2010. View Article : Google Scholar : PubMed/NCBI
32	McCawley LJ and Matrisian LM: Matrix metalloproteinases: Multifunctional contributors to tumor progression. Mol Med Today. 6:149–156. 2000. View Article : Google Scholar : PubMed/NCBI
33	Fang J, Shing Y, Wiederschain D, Yan L, Butterfield C, Jackson G, Harper J, Tamvakopoulos G and Moses MA: Matrix metal-loproteinase-2 is required for the switch to the angiogenic phenotype in a tumor model. Proc Natl Acad Sci USA. 97:3884–3889. 2000. View Article : Google Scholar
34	Rosenberger CM, Clark AE, Treuting PM, Johnson CD and Aderem A: ATF3 regulates MCMV infection in mice by modulating IFN-gamma expression in natural killer cells. Proc Natl Acad Sci USA. 105:2544–2549. 2008. View Article : Google Scholar : PubMed/NCBI
35	Stearns ME, Kim G, Garcia F and Wang M: Interleukin-10 induced activating transcription factor 3 transcriptional suppression of matrix metalloproteinase-2 gene expression in human prostate CPTX-1532 Cells. Mol Cancer Res. 2:403–416. 2004.PubMed/NCBI
36	Yan C, Wang H and Boyd DD: ATF3 represses 72-kDa type IV collagenase (MMP-2) expression by antagonizing p53-dependent trans-activation of the collagenase promoter. J Biol Chem. 277:10804–10812. 2002. View Article : Google Scholar : PubMed/NCBI