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Breast Cancer Research
Published in: Breast Cancer Research 1/2014

Open Access 01-02-2014 | Research article

Runx2 activates PI3K/Akt signaling via mTORC2 regulation in invasive breast cancer cells

Authors: Manish Tandon, Zujian Chen, Jitesh Pratap

Published in: Breast Cancer Research | Issue 1/2014

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Abstract

Introduction

The Runt-related transcription factor Runx2 is critical for skeletal development but is also aberrantly expressed in breast cancers, and promotes cell growth and invasion. A de-regulated serine/threonine kinase Akt signaling pathway is implicated in mammary carcinogenesis and cell survival; however, the mechanisms underlying Runx2 role in survival of invasive breast cancer cells are still unclear.

Methods

The phenotypic analysis of Runx2 function in cell survival was performed by gene silencing and flow cytometric analysis in highly invasive MDA-MB-231 and SUM-159-PT mammary epithelial cell lines. The expression analysis of Runx2 and pAkt (serine 473) proteins in metastatic breast cancer specimens was performed by immunohistochemistry. The mRNA and protein levels of kinases and phosphatases functional in Akt signaling were determined by real-time PCR and Western blotting, while DNA-protein interaction was studied by chromatin immunoprecipitation assays.

Results

The high Runx2 levels in invasive mammary epithelial cell lines promoted cell survival in Akt phosphorylation (pAkt-serine 473) dependent manner. The analysis of kinases and phosphatases associated with pAkt regulation revealed that Runx2 promotes pAkt levels via mammalian target of rapamycin complex-2 (mTORC2). The recruitment of Runx2 on mTOR promoter coupled with Runx2-dependent expression of mTORC2 component Rictor defined Runx2 function in pAkt-mediated survival of invasive breast cancer cells.

Conclusions

Our results identified a novel mechanism of Runx2 regulatory crosstalk in Akt signaling that could have important consequences in targeting invasive breast cancer-associated cell survival.
Appendix
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Literature
1.
Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin. 2011, 61: 69-90. 10.3322/caac.20107.CrossRefPubMed
2.
Engelman JA: Targeting PI3K signalling in cancer: opportunities, challenges and limitations. Nat Rev Cancer. 2009, 9: 550-562. 10.1038/nrc2664.CrossRefPubMed
3.
Ihle NT, Powis G: Take your PIK: phosphatidylinositol 3-kinase inhibitors race through the clinic and toward cancer therapy. Mol Cancer Ther. 2009, 8: 1-9.CrossRefPubMedPubMedCentral
4.
Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, Sato M, Okamoto R, Kitamura Y, Yoshiki S, Kishimoto T: Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 1997, 89: 755-764. 10.1016/S0092-8674(00)80258-5.CrossRefPubMed
5.
Pratap J, Imbalzano KM, Underwood JM, Cohet N, Gokul K, Akech J, van Wijnen AJ, Stein JL, Imbalzano AN, Nickerson JA, Lian JB, Stein GS: Ectopic runx2 expression in mammary epithelial cells disrupts formation of normal acini structure: implications for breast cancer progression. Cancer Res. 2009, 69: 6807-6814. 10.1158/0008-5472.CAN-09-1471.CrossRefPubMedPubMedCentral
6.
Onodera Y, Miki Y, Suzuki T, Takagi K, Akahira J, Sakyu T, Watanabe M, Inoue S, Ishida T, Ohuchi N, Sasano H: Runx2 in human breast carcinoma: its potential roles in cancer progression. Cancer Sci. 2010, 101: 2670-2675. 10.1111/j.1349-7006.2010.01742.x.CrossRefPubMed
7.
Brubaker KD, Vessella RL, Brown LG, Corey E: Prostate cancer expression of runt-domain transcription factor Runx2, a key regulator of osteoblast differentiation and function. Prostate. 2003, 56: 13-22. 10.1002/pros.10233.CrossRefPubMed
8.
Li H, Zhou RJ, Zhang GQ, Xu JP: Clinical significance of RUNX2 expression in patients with nonsmall cell lung cancer: a 5-year follow-up study. Tumour Biol. 2013, 34: 1807-1812. 10.1007/s13277-013-0720-4.CrossRefPubMed
9.
Li W, Xu S, Lin S, Zhao W: Overexpression of runt-related transcription factor-2 is associated with advanced tumor progression and poor prognosis in epithelial ovarian cancer. J Biomed Biotechnol. 2012, 2012: 456534-PubMedPubMedCentral
10.
Martin JW, Zielenska M, Stein GS, van Wijnen AJ, Squire JA: The role of RUNX2 in osteosarcoma oncogenesis. Sarcoma. 2011, 2011: 282745-CrossRefPubMed
11.
Nathan SS, Pereira BP, Zhou YF, Gupta A, Dombrowski C, Soong R, Pho RW, Stein GS, Salto-Tellez M, Cool SM, van Wijnen AJ: Elevated expression of Runx2 as a key parameter in the etiology of osteosarcoma. Mol Biol Rep. 2009, 36: 153-158. 10.1007/s11033-008-9378-1.CrossRefPubMed
12.
Pratap J, Lian JB, Stein GS: Metastatic bone disease: role of transcription factors and future targets. Bone. 2011, 48: 30-36. 10.1016/j.bone.2010.05.035.CrossRefPubMed
13.
Westendorf JJ: Transcriptional co-repressors of Runx2. J Cell Biochem. 2006, 98: 54-64. 10.1002/jcb.20805.CrossRefPubMed
14.
Chimge NO, Baniwal SK, Little GH, Chen YB, Kahn M, Tripathy D, Borok Z, Frenkel B: Regulation of breast cancer metastasis by Runx2 and estrogen signaling: the role of SNAI2. Breast Cancer Res. 2011, 13: R127-10.1186/bcr3073.CrossRefPubMedPubMedCentral
15.
Barnes GL, Javed A, Waller SM, Kamal MH, Hebert KE, Hassan MQ, Bellahcene A, van Wijnen AJ, Young MF, Lian JB, Stein GS, Gerstenfeld LC: Osteoblast-related transcription factors Runx2 (Cbfa1/AML3) and MSX2 mediate the expression of bone sialoprotein in human metastatic breast cancer cells. Cancer Res. 2003, 63: 2631-2637.PubMed
16.
Chimge NO, Frenkel B: The RUNX family in breast cancer: relationships with estrogen signaling. Oncogene. 2013, 32: 2121-2130. 10.1038/onc.2012.328.CrossRefPubMed
17.
Mendoza-Villanueva D, Deng W, Lopez-Camacho C, Shore P: The Runx transcriptional co-activator, CBFbeta, is essential for invasion of breast cancer cells. Mol Cancer. 2010, 9: 171-10.1186/1476-4598-9-171.CrossRefPubMedPubMedCentral
18.
Pratap J, Javed A, Languino LR, van Wijnen AJ, Stein JL, Stein GS, Lian JB: The Runx2 osteogenic transcription factor regulates matrix metalloproteinase 9 in bone metastatic cancer cells and controls cell invasion. Mol Cell Biol. 2005, 25: 8581-8591. 10.1128/MCB.25.19.8581-8591.2005.CrossRefPubMedPubMedCentral
19.
Pratap J, Lian JB, Javed A, Barnes GL, van Wijnen AJ, Stein JL, Stein GS: Regulatory roles of Runx2 in metastatic tumor and cancer cell interactions with bone. Cancer Metastasis Rev. 2006, 25: 589-600.CrossRefPubMed
20.
Pratap J, Wixted JJ, Gaur T, Zaidi SK, Dobson J, Gokul KD, Hussain S, van Wijnen AJ, Stein JL, Stein GS, Lian JB: Runx2 transcriptional activation of Indian Hedgehog and a downstream bone metastatic pathway in breast cancer cells. Cancer Res. 2008, 68: 7795-7802. 10.1158/0008-5472.CAN-08-1078.CrossRefPubMedPubMedCentral
21.
Bhaskar PT, Hay N: The two TORCs and Akt. Dev Cell. 2007, 12: 487-502. 10.1016/j.devcel.2007.03.020.CrossRefPubMed
22.
Guertin DA, Sabatini DM: Defining the role of mTOR in cancer. Cancer Cell. 2007, 12: 9-22. 10.1016/j.ccr.2007.05.008.CrossRefPubMed
23.
Zhang X, Tang N, Hadden TJ, Rishi AK: Akt, FoxO and regulation of apoptosis. Biochim Biophys Acta. 2011, 1813: 1978-1986. 10.1016/j.bbamcr.2011.03.010.CrossRefPubMed
24.
Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T, Clark L, Bayani N, Coppe JP, Tong F, Speed T, Spellman PT, DeVries S, Lapuk A, Wang NJ, Kuo WL, Stilwell JL, Pinkel D, Albertson DG, Waldman FM, McCormick F, Dickson RB, Johnson MD, Lippman M, Ethier S, Gazdar A, Gray JW: A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell. 2006, 10: 515-527. 10.1016/j.ccr.2006.10.008.CrossRefPubMedPubMedCentral
25.
Li LB, Toan SV, Zelenaia O, Watson DJ, Wolfe JH, Rothstein JD, Robinson MB: Regulation of astrocytic glutamate transporter expression by Akt: evidence for a selective transcriptional effect on the GLT-1/EAAT2 subtype. J Neurochem. 2006, 97: 759-771. 10.1111/j.1471-4159.2006.03743.x.CrossRefPubMed
26.
Pratap J, Galindo M, Zaidi SK, Vradii D, Bhat BM, Robinson JA, Choi JY, Komori T, Stein JL, Lian JB, Stein GS, van Wijnen AJ: Cell growth regulatory role of Runx2 during proliferative expansion of preosteoblasts. Cancer Res. 2003, 63: 5357-5362.PubMed
27.
Sarbassov DD, Guertin DA, Ali SM, Sabatini DM: Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science. 2005, 307: 1098-1101. 10.1126/science.1106148.CrossRefPubMed
28.
Wiznerowicz M, Trono D: Conditional suppression of cellular genes: lentivirus vector-mediated drug-inducible RNA interference. J Virol. 2003, 77: 8957-8961. 10.1128/JVI.77.16.8957-8951.2003.CrossRefPubMedPubMedCentral
29.
Tandon M, Gokul K, Ali SA, Chen Z, Lian J, Stein GS, Pratap J: Runx2 mediates epigenetic silencing of the bone morphogenetic protein-3B (BMP-3B/GDF10) in lung cancer cells. Mol Cancer. 2012, 11: 27-10.1186/1476-4598-11-27.CrossRefPubMedPubMedCentral
30.
Wingender E, Kel AE, Kel OV, Karas H, Heinemeyer T, Dietze P, Knuppel R, Romaschenko AG, Kolchanov NA: TRANSFAC, TRRD and COMPEL: towards a federated database system on transcriptional regulation. Nucleic Acids Res. 1997, 25: 265-268. 10.1093/nar/25.1.265.CrossRefPubMedPubMedCentral
31.
Shen H, Maki CG: Persistent p21 expression after Nutlin-3a removal is associated with senescence-like arrest in 4 N cells. J Biol Chem. 2010, 285: 23105-23114. 10.1074/jbc.M110.124990.CrossRefPubMedPubMedCentral
32.
Elstrom RL, Bauer DE, Buzzai M, Karnauskas R, Harris MH, Plas DR, Zhuang H, Cinalli RM, Alavi A, Rudin CM, Thompson CB: Akt stimulates aerobic glycolysis in cancer cells. Cancer Res. 2004, 64: 3892-3899. 10.1158/0008-5472.CAN-03-2904.CrossRefPubMed
33.
Huang H, Tindall DJ: Regulation of FOXO protein stability via ubiquitination and proteasome degradation. Biochim Biophys Acta. 2011, 1813: 1961-1964. 10.1016/j.bbamcr.2011.01.007.CrossRefPubMedPubMedCentral
34.
Liu Q, White LR, Clark SA, Heffner DJ, Winston BW, Tibbles LA, Muruve DA: Akt/protein kinase B activation by adenovirus vectors contributes to NFkappaB-dependent CXCL10 expression. J Virol. 2005, 79: 14507-14515. 10.1128/JVI.79.23.14507-14515.2005.CrossRefPubMedPubMedCentral
35.
Taniguchi CM, Emanuelli B, Kahn CR: Critical nodes in signalling pathways: insights into insulin action. Nat Rev Mol Cell Biol. 2006, 7: 85-96. 10.1038/nrm1837.CrossRefPubMed
36.
Gao T, Furnari F, Newton AC: PHLPP: a phosphatase that directly dephosphorylates Akt, promotes apoptosis, and suppresses tumor growth. Mol Cell. 2005, 18: 13-24. 10.1016/j.molcel.2005.03.008.CrossRefPubMed
37.
Lu Y, Muller M, Smith D, Dutta B, Komurov K, Iadevaia S, Ruths D, Tseng JT, Yu S, Yu Q, Nakhleh L, Balazsi G, Donnelly J, Schurdak M, Morgan-Lappe S, Fesik S, Ram PT, Mills GB: Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling. Oncogene. 2011, 30: 4567-4577. 10.1038/onc.2011.164.CrossRefPubMedPubMedCentral
38.
Klein PJ, Schmidt CM, Wiesenauer CA, Choi JN, Gage EA, Yip-Schneider MT, Wiebke EA, Wang Y, Omer C, Sebolt-Leopold JS: The effects of a novel MEK inhibitor PD184161 on MEK-ERK signaling and growth in human liver cancer. Neoplasia. 2006, 8: 1-8. 10.1593/neo.05373.CrossRefPubMedPubMedCentral
39.
Drissi H, Pouliot A, Stein JL, van Wijnen AJ, Stein GS, Lian JB: Identification of novel protein/DNA interactions within the promoter of the bone-related transcription factor Runx2/Cbfa1. J Cell Biochem. 2002, 86: 403-412. 10.1002/jcb.10238.CrossRefPubMed
40.
Park OJ, Kim HJ, Woo KM, Baek JH, Ryoo HM: FGF2-activated ERK mitogen-activated protein kinase enhances Runx2 acetylation and stabilization. J Biol Chem. 2010, 285: 3568-3574. 10.1074/jbc.M109.055053.CrossRefPubMed
41.
Jeon EJ, Lee KY, Choi NS, Lee MH, Kim HN, Jin YH, Ryoo HM, Choi JY, Yoshida M, Nishino N, Oh BC, Lee KS, Lee YH, Bae SC: Bone morphogenetic protein-2 stimulates Runx2 acetylation. J Biol Chem. 2006, 281: 16502-16511. 10.1074/jbc.M512494200.CrossRefPubMed
42.
Fujita T, Azuma Y, Fukuyama R, Hattori Y, Yoshida C, Koida M, Ogita K, Komori T: Runx2 induces osteoblast and chondrocyte differentiation and enhances their migration by coupling with PI3K-Akt signaling. J Cell Biol. 2004, 166: 85-95. 10.1083/jcb.200401138.CrossRefPubMedPubMedCentral
43.
Qiao M, Shapiro P, Fosbrink M, Rus H, Kumar R, Passaniti A: Cell cycle-dependent phosphorylation of the RUNX2 transcription factor by cdc2 regulates endothelial cell proliferation. J Biol Chem. 2006, 281: 7118-7128. 10.1074/jbc.M508162200.CrossRefPubMed
44.
Rajgopal A, Young DW, Mujeeb KA, Stein JL, Lian JB, van Wijnen AJ, Stein GS: Mitotic control of RUNX2 phosphorylation by both CDK1/cyclin B kinase and PP1/PP2A phosphatase in osteoblastic cells. J Cell Biochem. 2007, 100: 1509-1517. 10.1002/jcb.21137.CrossRefPubMed
45.
Kita K, Kimura T, Nakamura N, Yoshikawa H, Nakano T: PI3K/Akt signaling as a key regulatory pathway for chondrocyte terminal differentiation. Genes Cells. 2008, 13: 839-850. 10.1111/j.1365-2443.2008.01209.x.CrossRefPubMed
46.
Pande S, Browne G, Padmanabhan S, Zaidi SK, Lian JB, van Wijnen AJ, Stein JL, Stein GS: Oncogenic cooperation between PI3K/Akt signaling and transcription factor Runx2 promotes the invasive properties of metastatic breast cancer cells. J Cell Physiol. 2013, 28: 1784-1792.CrossRef
47.
Lengner CJ, Steinman HA, Gagnon J, Smith TW, Henderson JE, Kream BE, Stein GS, Lian JB, Jones SN: Osteoblast differentiation and skeletal development are regulated by Mdm2-p53 signaling. J Cell Biol. 2006, 172: 909-921. 10.1083/jcb.200508130.CrossRefPubMedPubMedCentral
48.
Pereira BP, Zhou Y, Gupta A, Leong DT, Aung KZ, Ling L, Pho RW, Galindo M, Salto-Tellez M, Stein GS, Cool SM, van Wijnen AJ, Nathan SS: Runx2, p53, and pRB status as diagnostic parameters for deregulation of osteoblast growth and differentiation in a new pre-chemotherapeutic osteosarcoma cell line (OS1). J Cell Physiol. 2009, 221: 778-788. 10.1002/jcp.21921.CrossRefPubMedPubMedCentral
49.
Mayo LD, Donner DB: A phosphatidylinositol 3-kinase/Akt pathway promotes translocation of Mdm2 from the cytoplasm to the nucleus. Proc Natl Acad Sci U S A. 2001, 98: 11598-11603. 10.1073/pnas.181181198.CrossRefPubMedPubMedCentral
50.
Ozaki T, Wu D, Sugimoto H, Nagase H, Nakagawara A: Runt-related transcription factor 2 (RUNX2) inhibits p53-dependent apoptosis through the collaboration with HDAC6 in response to DNA damage. Cell Death Dis. 2013, 4: e610-10.1038/cddis.2013.127.CrossRefPubMedPubMedCentral
51.
Yang S, Xu H, Yu S, Cao H, Fan J, Ge C, Fransceschi RT, Dong HH, Xiao G: Foxo1 mediates insulin-like growth factor 1 (IGF1)/insulin regulation of osteocalcin expression by antagonizing Runx2 in osteoblasts. J Biol Chem. 2011, 286: 19149-19158. 10.1074/jbc.M110.197905.CrossRefPubMedPubMedCentral
52.
Zhang H, Pan Y, Zheng L, Choe C, Lindgren B, Jensen ED, Westendorf JJ, Cheng L, Huang H: FOXO1 inhibits Runx2 transcriptional activity and prostate cancer cell migration and invasion. Cancer Res. 2011, 71: 3257-3267. 10.1158/0008-5472.CAN-10-2603.CrossRefPubMedPubMedCentral
53.
Blyth K, Vaillant F, Jenkins A, McDonald L, Pringle MA, Huser C, Stein T, Neil J, Cameron ER: Runx2 in normal tissues and cancer cells: A developing story. Blood Cells Mol Dis. 2010, 45: 117-123. 10.1016/j.bcmd.2010.05.007.CrossRefPubMed
54.
Lau QC, Raja E, Salto-Tellez M, Liu Q, Ito K, Inoue M, Putti TC, Loh M, Ko TK, Huang C, Bhalla KN, Zhu T, Ito Y, Sukumar S: RUNX3 is frequently inactivated by dual mechanisms of protein mislocalization and promoter hypermethylation in breast cancer. Cancer Res. 2006, 66: 6512-6520. 10.1158/0008-5472.CAN-06-0369.CrossRefPubMed
55.
Debnath J, Walker SJ, Brugge JS: Akt activation disrupts mammary acinar architecture and enhances proliferation in an mTOR-dependent manner. J Cell Biol. 2003, 163: 315-326. 10.1083/jcb.200304159.CrossRefPubMedPubMedCentral
56.
Wang L, Brugge JS, Janes KA: Intersection of FOXO- and RUNX1-mediated gene expression programs in single breast epithelial cells during morphogenesis and tumor progression. Proc Natl Acad Sci U S A. 2011, 108: E803-E812. 10.1073/pnas.1103423108.CrossRefPubMedPubMedCentral
57.
Edwards H, Xie C, LaFiura KM, Dombkowski AA, Buck SA, Boerner JL, Taub JW, Matherly LH, Ge Y: RUNX1 regulates phosphoinositide 3-kinase/AKT pathway: role in chemotherapy sensitivity in acute megakaryocytic leukemia. Blood. 2009, 114: 2744-2752. 10.1182/blood-2008-09-179812.CrossRefPubMedPubMedCentral
58.
Lin FC, Liu YP, Lai CH, Shan YS, Cheng HC, Hsu PI, Lee CH, Lee YC, Wang HY, Wang CH, Cheng JQ, Hsiao M, Lu PJ: RUNX3-mediated transcriptional inhibition of Akt suppresses tumorigenesis of human gastric cancer cells. Oncogene. 2012, 31: 4302-4316. 10.1038/onc.2011.596.CrossRefPubMed
59.
Pierce AD, Anglin IE, Vitolo MI, Mochin MT, Underwood KF, Goldblum SE, Kommineni S, Passaniti A: Glucose-activated RUNX2 phosphorylation promotes endothelial cell proliferation and an angiogenic phenotype. J Cell Biochem. 2012, 113: 282-292. 10.1002/jcb.23354.CrossRefPubMedPubMedCentral
60.
D’Souza DR, Salib MM, Bennett J, Mochin-Peters M, Asrani K, Goldblum SE, Renoud KJ, Shapiro P, Passaniti A: Hyperglycemia regulates RUNX2 activation and cellular wound healing through the aldose reductase polyol pathway. J Biol Chem. 2009, 284: 17947-17955. 10.1074/jbc.M109.002378.CrossRefPubMedPubMedCentral
Metadata
Title
Runx2 activates PI3K/Akt signaling via mTORC2 regulation in invasive breast cancer cells
Authors
Manish Tandon
Zujian Chen
Jitesh Pratap
Publication date
01-02-2014
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 1/2014
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3611

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