The RUNX Family in Breast Cancer: Relationships with Estrogen Signaling

REVIEW The RUNX family in breast cancer: relationships with estrogen signaling

N-O Chimge1 and B Frenkel2

The three RUNX family members are lineage specific master regulators, which also have important, context-dependent roles in carcinogenesis as either tumor suppressors or oncogenes. Here we review evidence for such roles in breast cancer (BCa). RUNX1, the predominant RUNX family member in breast epithelial cells, has a tumor suppressor role reflected by many somatic mutations found in primary tumor biopsies. The classical tumor suppressor gene RUNX3 does not consist of such a mutation hot spot, but it too seems to inhibit BCa; it is often inactivated in human BCa tumors and its haploinsufficiency in mice leads to spontaneous BCa development. The tumor suppressor activities of RUNX1 and RUNX3 are mediated in part by antagonism of estrogen signaling, a feature recently attributed to RUNX2 as well. Paradoxically, however RUNX2, a master osteoblast regulator, has been implicated in various aspects of metastasis in general and bone metastasis in particular. Reciprocating the anti-estrogenic tumor suppressor activity of RUNX proteins, inhibition of RUNX2 by estrogens may help explain their context-dependent anti-metastatic roles. Such roles are reserved to non-osseous metastasis, because ERa is associated with increased, not decreased skeletal dissemination of BCa cells. Finally, based on diverse expression patterns in BCa subtypes, the successful use of future RUNX-based therapies will most likely require careful patient selection.

Oncogene (2013) 32, 2121–2130; doi:10.1038/onc.2012.328; published online 8 October 2012 Keywords: RUNX1; RUNX2; RUNX3; tumor suppressor; oncogene; ERa

The mammalian RUNX family of transcription factors comprises diverse origins have been extensively reviewed,21–24 and Table 1 three members, RUNX1, RUNX2 and RUNX3, which have pivotal summarizes reported abnormalities implicating each of the RUNX roles in both normal development and neoplasia.1 Despite their proteins as a tumor suppressor in various cancer types. structural similarities, RUNX proteins have divergent physiological roles; RUNX1 is required for deﬁnitive hematopoiesis,2 RUNX2 has fundamental roles in the differentiation of osteoblasts and RUNX1 AS A BCa SUPPRESSOR chondrocytes,3–5 and RUNX3 is involved in gastrointestinal and 6–8 In addition to their most notable roles in hematopoietic cells, bone neuronal development. Inherited mutations in RUNX1 are the cells and neurons, the three RUNX genes are also expressed in the cause of familial platelet disorder with associated myeloid normal mammary gland, and their mRNA levels ﬂuctuate during malignancy,9 and RUNX2 mutations cause the dominant 25 10 cycles of pregnancy, lactation and involution. RUNX1 is the autosomal skeletal disease cleidocranial dysplasia. predominant RUNX family member expressed in human breast In cancer, RUNX act as either tumor suppressors or oncogenes 13,26,27 1,11 epithelial cells (Figure 1a) and the protein is readily detected depending on cellular context. RUNX1 is a frequent target in both the basal and the luminal cell layers.26 The most of translocations and other mutations in hematopoietic maligna- compelling evidence for its role in inhibiting BCa is the somatic ncies, and RUNX3, located on chromosome 1p36, resides mutations recently observed in RUNX1 (and not RUNX2 or RUNX3) in a deletion hotspot found in diverse cancers of epithelial, 28 12 in tumors from the Washington University BRC77 study and in hematopoietic and neuronal origins. Tumor suppressor activity the cohort of BCa patients studied in The Cancer Genome Atlas in breast cancer (BCa) has been attributed to all three RUNX pilot project29 (Figure 1b). Furthermore, RUNX1 was one of 17 proteins,13–15 whereas RUNX2 is increasingly recognized as pro- 30 11,16–20 genes whose expression pattern predicted BCa metastasis; its metastatic in breast (and prostate) cancer progression. Here expression is less abundant in BCa compared to normal breast we review the dual functions of RUNX proteins in BCa and their epithelial cells, and it progressively decreases with increasing relationships with estrogen signaling. BCa tumor aggression.31 Finally, experimental Ras-mediated transformation of basal-like MCF10A cells was associated with loss of RUNX1;31 and RUNX1 silencing in another study with RUNX PROTEINS ARE TUMOR SUPPRESSORS MCF10A cells led to the formation of abnormal, hyperproliferative All three mammalian RUNX family members have been implicated acinar structures in 3D cultures.13 in tumor suppression. The frequent functional inactivation of How RUNX1 suppresses BCa remains to be elucidated. Among RUNX1 and RUNX3 observed in hematopoietic and solid tumors of possible mechanisms, RUNX1 may stimulate E-cadherin to inhibit

1Department of Biochemistry and Molecular Biology, Institute for Genetic Medicine, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA and 2Departments of Orthopaedic Surgery and Biochemistry and Molecular Biology, Institute for Genetic Medicine, Keck School of Medicine of the University of Southern California, Los Angeles, CA, USA. Correspondence: Dr N-O Chimge or Dr B Frenkel, Department of Biochemistry and Molecular Biology, University of Southern California, Institute for Genetic Medicine, 2250 Alcazar Street, CSC-240, Los Angeles, CA 90033, USA. E-mail: [email protected] or [email protected] Received 3 May 2012; revised 20 June 2012; accepted 20 June 2012; published online 8 October 2012 RUNX1,2,3 in breast cancer N-O Chimge and B Frenkel 2122 Table 1. Tumor suppressor activities of the RUNX family in various cancers

RUNX Types of cancer Related mechanisms References

RUNX1 AML-M2 RUNX1-ETO: t(8;21)(q22;q22) 137–140 RUNX1-LRP16: t(11;21)(q13;q22) RUNX1-MTG16: t(16;21)(q24;q22) RUNX1-others Mut 141,142

MDS RUNX1-EVI: t(3;21)(q26.2;q22) 140,143–149 RUNX1-MDS1: t(3;21)(q26.2;q22) RUNX1-EAP: t(3;21)(q26.2;q22) RUNX1-PRDM6: t(1;21)(p36;q22) RUNX1-others Mut 150–156

B-ALL TEL-RUNX1: t(12;21)(p13;q22) 157–160 FPDMM Hemizygous deletion, SNP 9,161 AML-M1 Mut 162 AML-M0 LOH, biallelic RUNX1 mut 162–164 CMML Mut 165,166 Colorectal Mut 167 Gastointestinal tract Mut 168,169 Esophageal squamous-cell carcinoma SNP 170,171 Prostate SNP 172 Breast Mut 28,29

RUNX2 Osteosarcoma Inhibition of proliferation 173,174 Breast Antagonism of estrogen signaling 15

RUNX3 Gastric Hemizygous deletion, Mut, 8,167,169,175–177 hypermethylation, cytoplasmic mislocalization Bladder Mut, hypermethylation 178–180 Prostate Hypermethylation 181 Lung Hemizygous deletion, hypermethylation 182 Pancreatic Hemizygous deletion, hypermethylation 183 Colorectal Hypermethylation, Mut, cytoplasmic mislocalization 184–187 Brain Hypermethylation 188 Squamous cell carcinoma Cytoplasmic mislocalization 189 Hepatocellular carcinoma Hypermethylation 37,190 Bile duct Hemizygous deletion 183 Renal cell carcinoma Inhibition of proliferation 191 Bone Hypermethylation 192 Breast Hypermethylation, 14,34,35 cytoplasmic mislocalization Abbreviations: AML, acute myeloblastic leukemia; AML-M0, minimally differentiated acute myeloblastic leukemia; AML-M1, acute myeloblastic leukemia without maturation; CMML, chronic myelomonocytic leukemia; FPDMM, familial platelet disorder with associated myeloid malignancy; LOH, loss of heterozygosity; MDS, myelodysplastic syndrome; Mut, mutations other than SNPs or translocations; SNP, single-nucleotide polymorphism; t, translocation. Studies of breast cancer are in bold text.

epithelial-to-mesenchymal transition (EMT)32 and it may increased estradiol signaling owing to partial loss of ERa cooperate with FOXO signaling to protect ER-negative BCa cells destabilization by RUNX314 (Figure 2a). Expression of RUNX3 from oxidative stress.13 In ER-positive cells, however, we believe decreases during human BCa progression and high RUNX3 RUNX1 attenuates oncogenic effects of estrogens (Figure 2a) levels are associated with favorable prognosis.35 As in other mainly through physical interaction with ERa.33 In fact, all but one malignancies, RUNX3 is frequently inactivated in BCa by RUNX1 somatic mutations found so far are in ERa-positive promoter hypermethylation, genetic deletion, and protein tumors.28,29 mislocalization.34,35,37–42 RUNX3 promoter hypermethylation in response to estradiol has been observed in mammosphere- derived cells, which may contribute to the oncogenic activity of estradiol in breast epithelial cells.43 RUNX3 AS A BCa SUPPRESSOR Evidence suggesting a role for RUNX3 in inhibiting various malignancies is cited in Table 114,34–36 and the literature on the tumor suppressor role of RUNX3 speciﬁcally in BCa has been RUNX2 AS A BCa SUPPRESSOR recently reviewed.24 Most notably, RUNX3 hemizygous female RUNX2 is expressed in the developing mammary gland during mice develop spontaneous ductal carcinoma,14 attributable to embryogenesis5 and puberty, and its mRNA levels in glands of

Oncogene (2013) 2121 – 2130 & 2013 Macmillan Publishers Limited RUNX1,2,3 in breast cancer N-O Chimge and B Frenkel 2123 RUNX1: Chr.21q22.3 Scale: 200 kb hg19 RUNX1 RUNX1 RUNX1

Brain Heart Lymph Node Breast HMEC

RUNX2: Chr.6p21 Scale: 200 kb hg19 RUNX2 RUNX2 RUNX2

Brain Heart Lymph Node Breast HMEC

RUNX3: Chr.1p36 Scale: 200 kb hg19 RUNX3 RUNX3

Brain Heart Lymph Node Breast HMEC

frame shift deletion frame shift insertion RUNX1 in-frame deletion missense nonsense F259fs Q266* L288fs L317F 322e7+2 S100C L112fs 117e3+1 P125fs G135D N139S ELRNA143 in-frame del L161P R166Q G168R G168E R169fs R169K 169e4+1 D198G S218fs splice site splice site insertion

010050 150 200 250 300 350 400 450 481 Figure 1. RUNX1, highly expressed in breast epithelial cells, is frequently mutated in BCa. (a) RNA-sequencing data for RUNX1, RUNX2 and RUNX3 in human brain, heart, lymph node and breast tissues and in Human Mammary Epithelial Cells (HMEC) were produced by Wang et al.27 and displayed using the UCSC Genome Browser (http://genome.ucsc.edu). (b) Lollipop plot of RUNX1 somatic mutations identiﬁed by The Cancer Genome Atlas project29 and the Washington University BRC77 study.28 These studies indentiﬁed only one synonymous mutation in RUNX2 and none in RUNX3. Blue box represents the Runt DNA-binding domain.

5-week-old female mice are highest in the terminal end buds.44 resulted in a global increase of RUNX2 activity.15 Antagonism of Suggesting a physiological role in mammary epithelial cells, estradiol signaling by RUNX2 in vitro was particularly interesting RUNX2 regulates transcription of beta-casein,45 a mammary for a gene set, heretofore R2iEs, which was inhibited by RUNX2 gland-speciﬁc gene, and osteopontin,46 which is expressed in (R2i) and stimulated by estradiol (Es). The short-term response these cells during pregnancy and lactation. A growth-suppressive of the R2iEs gene set to letrozole treatment predicted role for RUNX2 was demonstrated in osteoblasts,47–49 and its clinical outcome better than other estradiol-regulated gene sets. tumor suppressor role in hormone carcinogenesis has been Thus, the R2iEs gene set, which is enriched for polo-like kinase- suggested based on the antagonism of ERa- and androgen related mitotic pathway genes,15 represents a critical growth receptor-mediated transcription in breast and prostate cancer regulatory node where the mitogenic property of estrogens cells, respectively.50,51 In the MCF7/Rx2dox BCa cell line, RUNX2 meets the anti-proliferative effect of RUNX2 in BCa cells. Finally, it diminished E2-mediated colony formation in soft agar.15 This was is important to note that RUNX2 can have either a negative attributable to the RUNX2-mediated general attenuation of the or a positive role in BCa initiation depending on cellular transcriptional response to E2, demonstrated by genome-wide context; contrasting its aforementioned tumor suppressor role in mRNA analysis.15 Consistent with this in vitro ﬁnding, inhibition the ERa-positive MCF7 BCa cell line,15 RUNX2 overexpression of estrogen signaling by letrozole treatment of BCa patients increased proliferation and induced abnormal acini formation

& 2013 Macmillan Publishers Limited Oncogene (2013) 2121 – 2130 RUNX1,2,3 in breast cancer N-O Chimge and B Frenkel 2124 similar to RUNX1, it cooperated with c-Myc,75 as well as with CBFb- SMMHC,76 to induce murine hematologic cancers. Amplification TUMOR TUMOR of RUNX2 on human chromosome 6p21 was related to INITIATION PROGRESSION osteosarcoma,77,78 a malignancy arising from osteoblasts, the ER ER cells in which RUNX2 has its most notable developmental role. Oncogenic Protective Furthermore, RUNX2 is overexpressed in carcinomas of the colon,79 pancreas80 and thyroid.81 In animal models of various malignancies including the prostate,82 lung83 and thyroid 84 RUNX RUNX2 cancer, an increase in RUNX2 observed early during cancer tumor suppressor pro-metastatic development likely induces EMT and local invasion. However, as discussed in detail below, most of the published work implicating RUNX2 in promoting cancer relates to prostate cancer and Figure 2. Working model for the reciprocal antagonism between 11,16–20,85–87 ERa and RUNX signaling. (a) During tumor initiation, RUNX proteins BCa. act as tumor suppressors in part by inhibiting ERa.(b) The beneficial The expression of bone-phenotypic genes in metastatic tumors effect of ERa in late-stage disease is attributable in part to repression was initially pursued in the context of prostate cancer, with the of the pro-metastatic properties of RUNX2. idea that the frequent dissemination of prostate cancer cells to bone may rely on osteomimetic mechanisms.88 A similar line of work followed in the context of BCa, which also metastasizes to bone with high frequency.89 Early studies documented ectopic expression in prostate cancer and BCa cells of CXCR4, MMPs, OPN, in the ERa-negative MCF10A cell line.17 Also contrasting its BSP, RANKL20,90–96 and finally RUNX2, the osteoblast master tumor suppressor activity is the pro-metastatic property regulator.97,98 RUNX2 was shown to prevent prostate cancer cell of RUNX2, which will be discussed in detail under the next apoptosis by stimulating the expression of survivin and Bcl2.82,99 subtitle. Increased levels of RUNX2 during prostate and breast tumor Multiple mechanisms could contribute to the antagonism of ERa growth was linked to BMP7-mediated protection of cancer cells by RUNX2. First, RUNX2 decreases ERa mRNA and protein levels in against apoptosis.100–102 Perturbations of RUNX2 in prostate BCa cells;15 this inhibition could occur at the transcriptional level cancer and BCa cells abolished their ability to form osteolytic because it has been shown, albeit in prostate cancer cells, that lesions in mouse xenograft model.16,20,103 RUNX2 is recruited to the ERa proximal promoter.52 Second, Recent studies have implicated RUNX2 in metastasis beyond a because RUNX2 binds ERa’s DNA-binding domain,51 the interaction role in bone homing, including local invasiveness as well as may result not only in decreased association of RUNX2 with its growth of tumors that have already metastasized to the skeleton. target genes,15,51 but also in decreased association of ERa with its Induction of RUNX2 in prostate cancer cells in vitro stimulated targets.15 Consistent with this idea, re-analysis of two recent data expression of numerous genes with known roles in EMT, sets33,53 indicated that genomic ERa targets are more likely to be invasiveness, extracellular matrix degradation, angiogenesis and antagonized by RUNX2 compared with non-genomic targets.15 osteolysis,11,15,17,19,20,86,104,105 with some of these metastatic Two observations suggest that this mechanism is likely specific for phenotypes possibly depending on RUNX2-stimulated protein RUNX2. First, in contrast to the interaction of RUNX2 with the DNA- secretion.52 Thus, whereas RUNX2 initiates an osteoblastic binding domain of ERa,51 the reported interaction with RUNX3 program in mesenchymal pluripotent cells,106 its ectopic involves the hinge region of ERa.24 Second, the C-terminus of expression in cancer cells initiates a transcriptional program RUNX2, which contains its ERa-binding region,51 is not conserved responsible for various aspects of metastasis, including but not in other RUNX proteins.1 Thus, whereas the BCa-suppressor limited to bone homing. activities of both RUNX2 and RUNX3 are at least in part Interestingly, manipulation of RUNX2 in prostate cancer and attributable to inhibition of ERa, the respective molecular BCa cells modulated the expression of several genes implicated mechanisms are likely distinct. in osteoclast differentiation and bone resorption, including OPG, CSF2, SPHK1, PTHrP, IL8 and several MMPs.15,17–20,104,107 Stimulation of bone resorption by prostate cancer cells is not ONCOGENIC PROPERTIES OF THE RUNX FAMILY AND THE unlike the increased resorption leading to the osteolytic lesions EMERGING ROLE OF RUNX2 IN METASTASIS typically associated with BCa bone metastasis, even though the Insertional mutagenesis studies in murine models have assigned co-stimulation of bone formation predominates in most cases of oncogenic properties to each of the three RUNX genes.54–62 prostate cancer bone metastases, resulting in osteoblastic Furthermore, RUNX163 synergized with c-Myc in murine tumors.108 It is the high bone turnover induced by both prostate lymphoma development, and acted as an oncogene in the cancer and BCa cells that elicits a vicious cycle whereby matrix- mouse skin by promoting epithelial cell proliferation.64 In humans, embedded growth factors, normally reserved to secure the 21q region that includes RUNX1 is amplified in a small, sufficient bone formation, are now released at higher rates and aggressive sub set of childhood B-cell lineage acute lymphoblastic hijacked to promote tumor cell growth.108,109 RUNX2-mediated leukemia.65–68 Moreover, RUNX1 was among the most highly tumor cell-driven osteoclastogenesis represents mimicry of up-regulated genes in invasive endometrial carcinoma.69–71 In BCa the osteoclastogenic role that RUNX2 has in the osteoblast RUNX1-mediated tethering of ERa to target genes33 may also lineage.110–115 Confirming the significance of RUNX2 in human contribute to breast hormone carcinogenesis. There is also some BCa aggression, Onodera et al.87 showed that RUNX2 immuno- evidence that RUNX3, a classical tumor suppressor, can act histochemical labeling index in primary tumor biopsies positively to promote human cancers. Possibly related to promoter correlated with disease progression and metastasis. The pro- demethylation, the gene is over-expressed in carcinomas of the metastatic properties of RUNX2 in the MCF7/Rx2dox BCa cell line head and neck,72 skin73 and ovary.74 critically depended on its target gene SNAI2, as knockdown of Whereas most of the cancer-related literature on RUNX genes, SNAI2 abolished RUNX2-driven EMT and invasiveness86 (Figure 3). in particular RUNX1 and RUNX3, concerns their tumor suppressor Conceivably, RUNX2-driven EMT in BCa cells86 recapitulates a role activities, RUNX2 is increasingly recognized in the cancer field for that RUNX2 has during normal branching morphogenesis, its oncogenic properties. RUNX2 was the most frequent target for as suggested by its co-expression with EMT-related genes in viral insertions in murine retroviral mutagenesis screens,61,68 and mammary gland terminal end buds.44

Oncogene (2013) 2121 – 2130 & 2013 Macmillan Publishers Limited RUNX1,2,3 in breast cancer N-O Chimge and B Frenkel 2125 ROLE OF RUNX2 AND ESTROGEN SIGNALING IN BCa As mentioned above, the osteomimetic property of prostate METASTASIS and later BCa was initially considered in the context of bone- Estrogen signaling, which promotes BCa initiation, has a homing. However, consistent with the role of RUNX2 in EMT paradoxical anti-metastatic role in advanced disease,116–118 and and invasion, which promote early metastatic events, the this is attributable in part to inhibiting the expression of RUNX2 predominant contribution of RUNX2 to the metastatic process is 86 target genes, including SNAI251,86,119 (Figure 3). Relationships not necessarily bone-specific. Other signaling pathways, such as among RUNX2, SNAI2 and ERa in vivo are demonstrable in gene Src, increase the predilection of a sub set of primary BCa cells to 120 expression profiles of BCa biopsies, where SNAI2 positively bone. These cells then give rise to metastases with a gene correlates with RUNX2 activity, and the RUNX2/SNAI2 axis expression signature that specifically reflects adaptation to 121 negatively correlates with ERa.86 Furthermore, in the study of bone, as opposed to the gene signature of BCa lung 122 Onodera et al., the negative correlation of RUNX2 with disease- metastases, which better reflects general metastatic properties. free survival was much stronger for ERa-negative compared with Because BCa metastases to bone versus other organs are ERa-positive tumors.87 Thus, antagonism between RUNX2 and processes with distinct molecular characteristics, it is not estrogen signaling in BCa has a role in two distinct contexts. First, surprising that they exhibit differential sensitivity to estrogen as discussed earlier, ERa signaling is targeted by RUNX2 (as well signaling. Whereas ERa is negatively correlated with non- 86 as RUNX1 and RUNX3), acting to suppress tumor initiation osseous metastasis, attributable in part to antagonism of the (Figure 2a). Second, the beneficial effect of ERa signaling in RUNX2/SNAI2 axis, it is positively correlated with bone 86,120,123 late-stage disease is attributable in part to inhibition of RUNX2 metastasis (Figure 3). Thus, ERa in bone-seeking tumors activity, now acting to promote metastasis (Figure 2b). not only permits expression of RUNX2 target genes, but in fact appears to promote bone metastasis. The co-existence of RUNX2 Primary and ERa in bone-seeking BCa cells may even result in positive Tumor interaction at regulatory elements of genes that promote bone- homing and growth within this microenvironment. Indeed, although RUNX2 and estrogen signaling are generally antagonis- RUNX2 tic, expression profiling of MCF7/Rx2dox cells also disclosed genes EMT, that are additively or synergistically regulated by the two 86 Invasion pathways. It will be interesting to test the role of these genes SNAI2 in bone metastasis and to compare the in vivo effects of RUNX2 on growth of ERa-positive versus ERa-negative BCa cells in bone ER versus non-osseous tissues.

RUNX PROTEINS IN BCa SUBTYPES BCa is a heterogeneous disease in terms of cell of origin, clinical SNAI2 RUNX2 progression and drug responsiveness. Based on their intrinsic gene expression profiles, BCa tumors can be classified into distinct ER molecular subtypes: luminal-like A, luminal-like B, basal-like, HER2- positive and normal breast-like.124,125 Figure 4 describes the Brain expression/activity of the three RUNX proteins, along with ERa and HER2, in a cohort of 557 tumors compiled previously86 and stratified per subtype based on the PAM-50 algorithm.126 Most Lung obviously, high RUNX2 activity and high RUNX3 expression characterize the ERa-negative basal-like tumors, attributing to the reported negative correlation between these two RUNX Bone Non-osseous members and ERa in BCa cells.14,15,24,43,51 High RUNX2 activity was metastasis metastasis also present in some HER2 enriched tumors, but not in the luminal subtypes, consistent with the positive correlation previously Figure 3. Roles of RUNX2 and ERa in BCa metastasis. According to 87 this working model, the RUNX2/SNAI2 axis promotes early meta- observed between RUNX2 and HER2. Finally, Figure 4 demon- static events (e.g., EMT, invasion, intravasation) and bone metastasis. strates a negative correlation between RUNX1 and RUNX2/3 and a ERa antagonizes RUNX2 during the early metastatic events but strong positive correlation between RUNX2 activity and RUNX3. operates along with the RUNX2/SNAI2 axis during bone metastasis. The interdependent expression of RUNX members in BCa (Figure 4)

Normal- HER2 like Luminal B Luminal A enriched Basal-like

2 HER2

ESR1

0 RUNX1

RUNX3

-2 RUNX2 metagene Standardized intensities Figure 4. RUNX genes in BCa subtypes. Each tumor from a heterogeneous cohort of 557 BCa patients86 was subtyped according to the PAM- 50 algorithm126 (courtesy of Dr Charles Perou). The values for RUNX1, RUNX3, HER2 and ESR1 represent the signal intensity associated with probe sets 210805_x_at,26 204197_s_at, 216836_s_at and 205225_at,120 respectively (Affymetrix, Santa Clara, CA, USA). Because of the ambiguity with the RUNX2 probe sets in the early Affymetrix arrays used to proﬁle mRNA expression, the RUNX2 values are those of a metagene deﬁned as the average normalized expression of the top 100 RUNX2-stimulated genes in MCF7/Rx2dox BCa cells.15

& 2013 Macmillan Publishers Limited Oncogene (2013) 2121 – 2130 RUNX1,2,3 in breast cancer N-O Chimge and B Frenkel 2126 is in line with the inverse relationships observed between RUNX1 review. This work was supported by NIH grant R01 DK071122 from the National and RUNX3 mRNA in B cells127,128 and RUNX1 and RUNX2 mRNAs Institute of Diabetes and Digestive and Kidney Diseases to BF, who holds the J Harold during skeletal development.129 These relationships are attributable and Edna L LaBriola Chair in Genetic Orthopaedic Research at USC. in part to the high similarities among the DNA-binding domains of the three RUNX proteins1 and the consequential similarities among REFERENCES their DNA binding elements52,130,131 and their transcriptomes.132 1 Blyth K, Cameron ER, Neil JC. The RUNX genes: gain or loss of function in cancer. 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RUNX2, sparing the tumor suppressor activities of RUNX1 and 18 Pratap J, Wixted JJ, Gaur T, Zaidi SK, Dobson J, Gokul KD et al. Runx2 tran- RUNX3. During the development of such drugs, special attention scriptional activation of Indian Hedgehog and a downstream bone metastatic should be paid to the potential loss of the inhibition of estrogen pathway in breast cancer cells. Cancer Res 2008; 68: 7795–7802. signaling in ERa-positive tumors. On the other hand, the design of 19 Baniwal SK, Khalid O, Gabet Y, Shah RR, Purcell DJ, Mav D et al. Runx2 tran- future selective estrogen receptor modulators (SERMs) should take scriptome of prostate cancer cells: insights into invasiveness and bone metastasis. Mol Cancer 2010; 9: 258. into consideration their effects on RUNX proteins, with the ideal 20 Akech J, Wixted JJ, Bedard K, van der Deen M, Hussain S, Guise TA et al. Runx2 SERM possessing both anti-ER and anti-RUNX2, but not anti- association with progression of prostate cancer in patients: mechanisms med- RUNX1 or anti-RUNX3 properties. iating bone osteolysis and osteoblastic metastatic lesions. Oncogene 2010; 29: 811–821. 21 Chuang LS, Ito Y. RUNX3 is multifunctional in carcinogenesis of multiple solid CONFLICT OF INTEREST tumors. Oncogene 2010; 29: 2605–2615. 22 Mangan JK, Speck NA. RUNX1 mutations in clonal myeloid disorders: from The authors declare no conflict of interest. conventional cytogenetics to next generation sequencing. A story 40 years in the making. Crit Rev Oncog 2011; 16: 77–91. 23 De Braekeleer E, Ferec C, De Braekeleer M. RUNX1 translocations in malignant ACKNOWLEDGEMENTS hemopathies. Anticancer Res 2009; 29: 1031–1037. We thank Gerhard A Coetzee, Debu Tripathy, Gillian H Little (University of Southern 24 Chen LF. Tumor suppressor function of RUNX3 in breast cancer. J Cell Biochem California) and Andre van Wijnen (University of Massachusetts Medical School) for 2012; 113: 1470–1477. carefully reading the manuscript and providing critical comments. We are particularly 25 Blyth K, Vaillant F, Jenkins A, McDonald L, Pringle MA, Huser C et al. Runx2 thankful to Charles Perou (University of North Carolina), Dan Koboldt and Matthew in normal tissues and cancer cells: a developing story. Blood Cells Mol Dis 2010; Ellis (Washington University), who also shared, analyzed and edited data for this 45: 117–123.

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