Twist Is Transcriptionally Induced by Activation of STAT3 and Mediates STAT3 Oncogenic Function

To explore the basis of metastasis, we compared the human breast cancer lines MCF-7 and MDA-MB453, which have low invasive ability, with their sublines MCF7-I4 and MDA-MB453-I4 with high invasive ability for gene expression and signaling pathways. We previously showed that the I4 lines had dramatically elevated levels of Twist compared with their parental lines. In this study, we observed significantly increased STAT3 Tyr705 phosphorylation, but not the STAT3 protein levels, in the I4 lines. Activation of STAT3 by interleukin-6 or expression of activated Src induced Twist expression at protein and mRNA levels. Inhibiting STAT3 by a small molecule inhibitor, JSI-124, STAT3 small hairpin RNAs, or dominant negative STAT3 resulted in significant reduction of Twist protein and mRNA expression. STAT3 directly bound to the second proximal STAT3-binding site on the human Twist promoter and activated its transcriptional activity. Inhibition of STAT3 reduced migration, invasion, and colony formation of the I4 cells. Ectopic expression of Twist significantly rescued those phenotypes. Ten normal and 46 tumor specimens of breast tissues were examined for activation of STAT3 and expression of Twist. There was a strong correlation between Tyr705 p-STAT3 and Twist level in the late stage tumor tissues. Our results indicate that activated STAT3 transcriptionally induces Twist, which plays an important role in promoting migration, invasion, and anchorage-independent growth. Together with our previous observation that Twist transcriptionally induces AKT2 to mediate Twist-promoted oncogenic functions, we conclude that STAT3, Twist, and AKT2 form a functional signaling axis to regulate pivotal oncogenic properties of cancer cells. To explore the basis of metastasis, we compared the human breast cancer lines MCF-7 and MDA-MB453, which have low invasive ability, with their sublines MCF7-I4 and MDA-MB453-I4 with high invasive ability for gene expression and signaling pathways. We previously showed that the I4 lines had dramatically elevated levels of Twist compared with their parental lines. In this study, we observed significantly increased STAT3 Tyr705 phosphorylation, but not the STAT3 protein levels, in the I4 lines. Activation of STAT3 by interleukin-6 or expression of activated Src induced Twist expression at protein and mRNA levels. Inhibiting STAT3 by a small molecule inhibitor, JSI-124, STAT3 small hairpin RNAs, or dominant negative STAT3 resulted in significant reduction of Twist protein and mRNA expression. STAT3 directly bound to the second proximal STAT3-binding site on the human Twist promoter and activated its transcriptional activity. Inhibition of STAT3 reduced migration, invasion, and colony formation of the I4 cells. Ectopic expression of Twist significantly rescued those phenotypes. Ten normal and 46 tumor specimens of breast tissues were examined for activation of STAT3 and expression of Twist. There was a strong correlation between Tyr705 p-STAT3 and Twist level in the late stage tumor tissues. Our results indicate that activated STAT3 transcriptionally induces Twist, which plays an important role in promoting migration, invasion, and anchorage-independent growth. Together with our previous observation that Twist transcriptionally induces AKT2 to mediate Twist-promoted oncogenic functions, we conclude that STAT3, Twist, and AKT2 form a functional signaling axis to regulate pivotal oncogenic properties of cancer cells. Twist, a highly conserved basic helix-loop-helix transcriptional factor, was previously shown to play a pivotal role in mesodermal, myoblast, and osteoblast differentiation (1Bialek P. Kern B. Yang X. Schrock M. Sosic D. Hong N. Wu H. Yu K. Ornitz D.M. Olson E.N. Justice M.J. Karsenty G. Dev. Cell. 2004; 6: 423-435Abstract Full Text Full Text PDF PubMed Scopus (552) Google Scholar, 2Baylies M.K. Bate M. Science. 1996; 272: 1481-1484Crossref PubMed Scopus (285) Google Scholar, 3Castanon I. Von Stetina S. Kass J. Baylies M.K. Development. 2001; 128: 3145-3159Crossref PubMed Google Scholar). Mutational inactivation of the Twist gene resulted in Saethre-Chotzen syndrome, an autosomal dominant disorder characterized by premature fusion of the cranial sutures, skull deformations, limb abnormalities, and facial dysmorphisims (4El Ghouzzi V. Legeai-Mallet L. Aresta S. Benoist C. Munnich A. de Gunzburg J. Bonaventure J. Hum. Mol. Genet. 2000; 9: 813-819Crossref PubMed Scopus (78) Google Scholar). Recent studies have demonstrated that Twist also played a key role in the development and progression of human cancer (5Puisieux A. Valsesia-Wittmann S. Ansieau S. Br. J. Cancer. 2006; 94: 13-17Crossref PubMed Scopus (181) Google Scholar). Twist is frequently overexpressed in human rhabdomyosarcoma, gastric carcinoma, melanoma, breast cancer, prostate cancer, liver carcinoma, and glioma (5Puisieux A. Valsesia-Wittmann S. Ansieau S. Br. J. Cancer. 2006; 94: 13-17Crossref PubMed Scopus (181) Google Scholar). The elevated Twist protein levels are associated with advanced tumor stage and poor prognosis in several types of cancer (6Zhang Z. Xie D. Li X. Wong Y.C. Xin D. Guan X.Y. Chua C.W. Leung S.C. Na Y. Wang X. Hum. Pathol. 2007; 38: 598-606Crossref PubMed Scopus (98) Google Scholar, 7Ohuchida K. Mizumoto K. Ohhashi S. Yamaguchi H. Konomi H. Nagai E. Yamaguchi K. Tsuneyoshi M. Tanaka M. Int. J. Cancer. 2007; 120: 1634-1640Crossref PubMed Scopus (61) Google Scholar, 8Lee T.K. Poon R.T. Yuen A.P. Ling M.T. Kwok W.K. Wang X.H. Wong Y.C. Guan X.Y. Man K. Chau K.L. Fan S.T. Clin. Cancer Res. 2006; 12: 5369-5376Crossref PubMed Scopus (361) Google Scholar). Further, increased Twist in cancer cells has been shown to promote metastatic ability in vivo and induce epithelial to mesenchymal transition, cell survival, angiogenesis, and chemoresistance in vitro (9Yang J. Mani S.A. Donaher J.L. Ramaswamy S. Itzykson R.A. Come C. Savagner P. Gitelman I. Richardson A. Weinberg R.A. Cell. 2004; 117: 927-939Abstract Full Text Full Text PDF PubMed Scopus (3156) Google Scholar, 10Maestro R. Dei Tos A.P. Hamamori Y. Krasnokutsky S. Sartorelli V. Kedes L. Doglioni C. Beach D.H. Hannon G.J. Genes Dev. 1999; 13: 2207-2217Crossref PubMed Scopus (462) Google Scholar, 11Mironchik Y. Winnard Jr., P.T. Vesuna F. Kato Y. Wildes F. Pathak A.P. Kominsky S. Artemov D. Bhujwalla Z. Van Diest P. Burger H. Glackin C. Raman V. Cancer Res. 2005; 65: 10801-10809Crossref PubMed Scopus (246) Google Scholar, 12Zong C.S. Zeng L. Jiang Y. Sadowski H.B. Wang L.H. J. Biol. Chem. 1998; 273: 28065-28072Abstract Full Text Full Text PDF PubMed Scopus (92) Google Scholar, 13Yuen H.F. Chan Y.P. Wong M.L. Kwok W.K. Chan K.K. Lee P.Y. Srivastava G. Law S.Y. Wong Y.C. Wang X. Chan K.W. J. Clin. Pathol. 2006; 60: 510-514Crossref PubMed Scopus (89) Google Scholar, 14Pham C.G. Bubici C. Zazzeroni F. Knabb J.R. Papa S. Kuntzen C. Franzoso G. Mol. Cell Biol. 2007; 27: 3920-3935Crossref PubMed Scopus (116) Google Scholar). In addition, ectopic expression of Twist in mouse embryonic fibroblasts promotes soft agar colony formation, indicating its role in malignant transformation (10Maestro R. Dei Tos A.P. Hamamori Y. Krasnokutsky S. Sartorelli V. Kedes L. Doglioni C. Beach D.H. Hannon G.J. Genes Dev. 1999; 13: 2207-2217Crossref PubMed Scopus (462) Google Scholar). A number of downstream target genes of Twist have been identified and shown to mediate its function (10Maestro R. Dei Tos A.P. Hamamori Y. Krasnokutsky S. Sartorelli V. Kedes L. Doglioni C. Beach D.H. Hannon G.J. Genes Dev. 1999; 13: 2207-2217Crossref PubMed Scopus (462) Google Scholar). Previous studies have demonstrated that epithelial to mesenchymal transition-associated molecules, such as E-cadherin and N-cadherin, are tightly regulated by Twist (9Yang J. Mani S.A. Donaher J.L. Ramaswamy S. Itzykson R.A. Come C. Savagner P. Gitelman I. Richardson A. Weinberg R.A. Cell. 2004; 117: 927-939Abstract Full Text Full Text PDF PubMed Scopus (3156) Google Scholar, 15Alexander N.R. Tran N.L. Rekapally H. Summers C.E. Glackin C. Heimark R.L. Cancer Res. 2006; 66: 3365-3369Crossref PubMed Scopus (200) Google Scholar). We have recently shown that AKT signaling is promoted by Twist through transcriptional up-regulation of AKT2 (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google Scholar). However, the molecular mechanism for the up-regulation of Twist in cancer cells is less clear. Signal transducer and activator of transcription 3 (STAT3) 4The abbreviations used are: STAT, signal transducers and activators of transcription; IL, interleukin; shRNA, small hairpin RNA; DMEM, Dulbecco's modified Eagle's medium; FCS, fetal calf serum; RT, reverse transcription; ChIP, chromatin immunoprecipitation; EMSA, electrophoretic mobility shift assay; GFP, green fluorescent protein; ERK, extracellular signal-regulated kinase; MMP, matrix metalloproteinase. protein is a member of a family of latent cytoplasmic transcriptional factors that convey signals from the cell surface to the nucleus upon activation by cytokines and growth factors (17Haura E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar). Engagement of cell surface receptors by polypeptide ligands induces tyrosine phosphorylation of STAT3 protein by Janus kinases, growth factor receptor tyrosine kinases, and, in some cases, Src family tyrosine kinases. The phospho-STAT3 protein dimerizes and translocates to nucleus to regulate expression of the genes harboring STAT3-binding sites in their promoters (18Levy D.E. Darnell Jr., J.E. Nat. Rev. Mol. Cell Biol. 2002; 3: PubMed Scopus Google Scholar). STAT3 has been shown to regulate genes that cell survival, and development (18Levy D.E. Darnell Jr., J.E. Nat. Rev. Mol. Cell Biol. 2002; 3: PubMed Scopus Google Scholar). STAT3 was to cells in vitro D. Darnell Jr., J.E. Mol. Cell Biol. 1998; PubMed Scopus Google and is for cell transformation of a number of C.S. Zeng L. Jiang Y. Sadowski H.B. Wang L.H. J. Biol. Chem. 1998; 273: 28065-28072Abstract Full Text Full Text PDF PubMed Scopus (92) Google Scholar, D. Darnell Jr., J.E. Mol. Cell Biol. 1998; PubMed Scopus Google Scholar, C. J. Jove R. Science. PubMed Scopus Google Scholar). studies have STAT3 in human tumor specimens S. 2005; PubMed Scopus Google Scholar, L. A. J. J. Clin. 2004; PubMed Scopus Google Scholar, A. Int. J. Cancer. 1999; PubMed Scopus Google Scholar, R. Yu A. R. K.L. Jove R. Cell Google Scholar). STAT3 in tumor cells in expression of genes in cell and V. M. V. M. M. N. A. J. Cell PubMed Scopus Google Scholar). We previously a to highly invasive cells from a of low invasive breast cancer lines and showed an increased level of Twist in the highly invasive cells (17Haura E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar). We demonstrated that Twist transcriptionally induced AKT2 to mediate migration, invasion, and (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google Scholar). In the study, we an elevated level of activated STAT3 in the highly invasive cells and demonstrated its role in transcriptional of Twist. We showed that STAT3 bound to the Twist promoter and activated its transcriptional activity. Twist was to the oncogenic from of Further, elevated p-STAT3 and Twist are highly in breast Our for the that Twist is a transcriptional target of Together with our previous (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google we conclude that STAT3, Twist, and AKT2 are in a signaling to regulate oncogenic properties of breast cancer cells. Cell and breast cancer cell lines MCF-7 and and their highly invasive cell lines and have been as previously (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google Scholar). The cells were in Dulbecco's modified Eagle's with fetal calf and The were in a at in the of The were with to the human breast cancer specimens were in the H. Lee Cancer the The tissues were and at and STAT3 and STAT3 were from Cell and were from and (17Haura E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar). and growth factor reduced were from and STAT3 was from and of the Twist human Twist promoter was by from the human cell The used for the Twist promoter were and Twist promoter and its were by a second of Full 3 and The were with and The were by Twist was previously (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google Scholar). was as previously C.S. Zeng L. Jiang Y. Sadowski H.B. Wang L.H. J. Biol. Chem. 1998; 273: 28065-28072Abstract Full Text Full Text PDF PubMed Scopus (92) Google Scholar). the cells were with and and the was by and with as in the and was and was with The was with reverse of and was with gene for as an and Twist Cell and were in a with for and as (17Haura E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar). The were on with of growth factor reduced at cells were in of the The was with of for cells in the were with The cells on the were with a for The cells at of the were was of and of cells were and was a and were on a was as C.S. Zeng L. Jiang Y. Sadowski H.B. Wang L.H. J. Biol. Chem. 1998; 273: 28065-28072Abstract Full Text Full Text PDF PubMed Scopus (92) Google Scholar, E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar). cells were in The cells were a a agar of and The were to the and with of normal growth of were and by cells were in a and with and as in the The of in was by the of the cell were and and were to the The of cell was used for the and the was to that of was as or was in was as previously with J. A. R. F. S. A. G. J. M. Van J.L. M. Mol. Cell Biol. PubMed Scopus Google Scholar). chromatin was from a of cells that were with STAT3 and The chromatin was with and and with protein A with of and The chromatin was and in with an or an the was from the by the in at for at protein and were by with of and of A at for 3 was to with for the STAT3-binding sites of the transcriptional The of the used are: proximal and and The proximal and the proximal and STAT3-binding cells were with of was as previously S. 2005; PubMed Scopus Google Scholar). of protein were with The the STAT3-binding sites in the Twist promoter and their are as the second STAT3-binding the STAT3 were by and by were on the from the of the with a hairpin was at and sites from of are as and A of and a that protein were used to and cells. STAT3 cells were by of and protein an of the tissues was and an of was and the tissues was to The were of protein from was by was as previously (17Haura E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google and were used for the the were with for to activity. in the were with the in at and with The signal was by formation and with by with in and and STAT3 in the I4 and STAT3 Activation Twist at and mRNA the molecular mechanism of breast cancer metastasis, we previously highly invasive lines and by of MCF-7 and and showed that AKT2 and Twist were elevated in the invasive cells compared with the parental cells. Further, we demonstrated that Twist bound to the AKT2 promoter and induced AKT2 expression (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google Scholar). However, the basis for up-regulation of Twist in I4 cells is activation of STAT3 in late stage and high breast cancer to STAT3 is activated in the I4 cells. that the levels of STAT3, a activation upon its phosphorylation, were increased in and the levels of STAT3 not as compared with their parental cells STAT3 and Twist to tumor we the that STAT3 and Twist To this and cells were with the small molecule of STAT3, Sun J. A. Turkson J. Jove R. Cancer Res. Google Scholar). with for we observed reduced phospho-STAT3 levels and Twist protein expression that Twist protein level is by the of STAT3 To the of Twist by STAT3, we of STAT3 in the I4 cells. and MDA-MB453-I4 cells were with a or a of of shown in the level of Twist protein with STAT3 The I4 cells showed a reduction of Twist mRNA as by to that Twist is a downstream target gene of Activation of STAT3 Twist our we STAT3 which is to and STAT3 C. J. Jove R. Science. PubMed Scopus Google Scholar, X. A. Mol. Cell Biol. 1996; PubMed Scopus Google in cells and for Twist expression. of STAT3 not an in Twist protein expression of Src or of and STAT3 to a significant in phospho-STAT3 and Twist levels, with the a level The mRNA level of Twist was examined by Ectopic expression of increased Twist and a was STAT3 was The of on Twist levels by the expression of a dominant negative STAT3 to Z. Darnell Jr., J.E. Genes Dev. 9: PubMed Scopus Google that this was a We this observed activation of To we examined Twist protein levels in and its normal shown in cells had elevated and Twist levels as compared with those of the cells. phosphorylation was used as an of and as a significantly level of was in cells. the Src family protein tyrosine STAT3 also activated by has been shown to a STAT3 activation signal J. C. F. Mol. Cell Biol. 13: PubMed Scopus Google Scholar). To the of Twist by STAT3 we cell and and cells with or with STAT3 resulted in increased p-STAT3 as as Twist at protein and mRNA levels, and the was in the of and our results indicate that Twist expression is regulated by STAT3, that up-regulation of Twist in the I4 cells and tumor cells to the activation of STAT3 to and the Twist and the of human and mouse Twist we identified the human Twist promoter and sites to the STAT3-binding (17Haura E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar). we the human Twist promoter the A was to activated STAT3 was to the human Twist of and of STAT3 to a of Twist promoter the Twist promoter activity. the of induced the promoter which was by STAT3 from activation of STAT3, of and STAT3 to a of Twist promoter which was to the level upon of dominant negative STAT3 results the that activated STAT3 regulate Twist promoter those to and human Twist A and cells were with and as as the or with of and with or was and to activity. The was in in a of to The results are the of in a of of the Twist promoter were cells with or and and of in a with or and the cells were to as of were with from MCF7-I4 cells. was examined by of the with and cells were with STAT3 and and and MCF7-I4 cells were with or as or chromatin was or as a negative The were to as used as for the of is a and the not the Twist promoter STAT3-binding we which to of the human Twist promoter were on the of the STAT3-binding showed that the second STAT3-binding on the Twist promoter proximal to the transcriptional site is for the activation of the promoter In addition, to have a between and of the promoter 3 and To STAT3 the Twist promoter in vitro and in we and showed that the second but not the STAT3-binding site directly bound to STAT3 A was of that the STAT3-binding The of the proximal STAT3-binding the second of the STAT3-binding shown in the proximal Twist promoter in the chromatin with an to the STAT3 sites of the Twist promoter not promoter in the was in cells that were with or and not the second proximal STAT3 site the Twist promoter the and sites not the promoter our that the second proximal site is the STAT3 site is of the previously by Wang F. PubMed Scopus (89) Google Scholar). our that STAT3 directly the Twist promoter and its Twist and upon of the transcriptional mechanism of Twist we to the of the in the of anchorage-independent migration, and The of STAT3 in with to a in colony formation ability, which was rescued by the of Twist the that Twist is downstream of STAT3 and mediate STAT3 oncogenic In addition, we the of axis on cell and cells were with the STAT3 or and examined for and Inhibition of STAT3 activation significantly and of the I4 which were but significantly rescued by Twist of STAT3 also cell and this not have significant on our cell and the number of and cells was used the that the axis is an important in and anchorage-independent growth and that STAT3 its at in through Twist. of Twist with STAT3 Activation in of observed that STAT3 Twist expression in cancer we this in We examined 46 breast and normal specimens for phospho-STAT3 and Twist expression by and A and the 46 breast had elevated phospho-STAT3 levels, and showed of Twist. the with elevated also had elevated Twist levels, 3 had low Twist levels of phospho-STAT3 and Twist are to the nucleus of the cancer cells and not to that of the cells which have had elevated phospho-STAT3 levels, and had level of Twist expression elevated levels of Twist were not significantly between the tumor increased Twist expression was associated with late stage of breast Further, of Twist and p-STAT3 was in late stage of the of and functional between STAT3 and Twist. our invasive breast cancer (16Cheng G.Z. Chan J. Wang Q. Zhang W. Sun C.D. Wang L.H. Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google we demonstrated a between activated STAT3 and Twist, key transcriptional factors that are associated with breast cancer The correlation of STAT3 and Twist levels has been observed the of Twist level was regulated by STAT3 were not X. Cancer Res. 2005; 65: PubMed Scopus Google Scholar). Our that STAT3 is a factor for up-regulation of Twist at the transcriptional Our that Twist is a target gene of STAT3, and signaling plays an important role in and growth of breast cancer cells. Our showed that with resulted in of growth at of and anchorage-independent to the observed of colony formation upon with Our the that Twist is transcriptionally regulated by STAT3 and STAT3 of Twist as a transcriptional target and of the STAT3-binding site on Twist promoter is in with that observed by S.C. W. X. Y. J.L. Cancer Res. 2007; 67: PubMed Scopus Google Scholar). Together with our previous that Twist transcriptionally a signaling axis in breast cancer progression and also at in the of in cancer cells. A number of have demonstrated that Twist is frequently overexpressed in a of types of human and is associated with late stage and high However, of Twist at the level the tumor In the study, we showed of activated STAT3 and of Twist in human breast of phospho-STAT3 and elevated Twist expression in breast the of the signaling is that activated STAT3 is of the transcriptional factors that to up-regulation of Twist in human transcriptional of Twist is C.G. Bubici C. Zazzeroni F. Knabb J.R. Papa S. Kuntzen C. Franzoso G. Mol. Cell Biol. 2007; 27: 3920-3935Crossref PubMed Scopus (116) Google its role in Twist in cancer is The of STAT3 in has been by its phosphorylation in the of human and its to induce cell transformation D. Darnell Jr., J.E. Mol. Cell Biol. 1998; PubMed Scopus Google Scholar, K. M. R. K. S. W. 2002; Full Text Full Text PDF PubMed Scopus Google Scholar). STAT3 and its downstream are in of from to progression to (9Yang J. Mani S.A. Donaher J.L. Ramaswamy S. Itzykson R.A. Come C. Savagner P. Gitelman I. Richardson A. Weinberg R.A. Cell. 2004; 117: 927-939Abstract Full Text Full Text PDF PubMed Scopus (3156) Google Scholar, E.B. Turkson J. Jove R. Nat. Clin. Pract. Oncol. 2005; 2: 315-324Crossref PubMed Scopus (375) Google Scholar, R. Jove R. Clin. Cancer Res. 2002; Google Scholar). The invasion, is of the in cancer in the of the of cancer cells. that STAT3 is in cell migration, and in and normal STAT3 cell up-regulation of the gene S. C. A. R. Dev. Cell. 2002; 2: Full Text Full Text PDF PubMed Scopus Google Scholar, S. C. N. 2004; PubMed Scopus Google a for A. I. A. M.J. F. Nat. Cell Biol. 2000; 2: PubMed Scopus Google Scholar). In of cell migration, in cell as in of phosphorylation M. S. S. K. J. S. J. Biol. 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Cancer Res. 2007; 67: 1979-1987Crossref PubMed Scopus (465) Google Scholar). on our we that STAT3 regulate AKT2 expression through Twist. a previous has demonstrated that STAT3 transcriptionally expression V. Y. Mol. Cell. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). that STAT3 promote cell and tumor Twist and that STAT3 or its downstream Twist and AKT2 to of tumor progression and the of the axis as an target for cancer with