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NMIIA promotes tumor growth and metastasis by activating the Wnt/β-catenin signaling pathway and EMT in pancreatic cancer

Oncogene (2019) | Download Citation

Abstract

Non-muscle myosin IIA (NMIIA) protein plays an important role in cell cytokinesis and cell migration. The role and underlying regulatory mechanisms of NMIIA in pancreatic cancer (PC) remain elusive. We found that NMIIA is highly expressed in PC tissues and contributes to PC poor progression by using open microarray datasets from the Gene Expression Omnibus (GEO), The Cancer Genome Atlas (TCGA), and PC tissue arrays. NMIIA regulates β-catenin mediated EMT to promote the proliferation, migration, invasion, and sphere formation of PC cells in vitro and in vivo. NMIIA controls the β-catenin transcriptional activity by interacting with β-catenin. Moreover, MEK/ERK signaling is critical in MLC2 (Ser19) phosphorylation, which can mediate NMIIA activity and regulate Wnt/β-catenin signaling. These findings highlight the significance of NMIIA in tumor regression and implicate NMIIA as a promising candidate for PC treatment.

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References

  1. 1.

    Brower V. Genomic research advances pancreatic cancer’s early detection and treatment. J Natl Cancer Inst. 2015;107:djv195. pii

  2. 2.

    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2015. CA Cancer J Clin. 2015;65:5–29.

  3. 3.

    Iacobuzio-Donahue CA, Fu B, Yachida S, Luo M, Abe H, Henderson CM, et al. DPC4 gene status of the primary carcinoma correlates with patterns of failure in patients with pancreatic cancer. J Clin Oncol. 2009;27:1806–13.

  4. 4.

    Ryan DP, Hong TS, Bardeesy N. Pancreatic adenocarcinoma. N Engl J Med. 2014;371:2140–1.

  5. 5.

    Brabletz T. To differentiate or not-routes towards metastasis. Nat Rev Cancer. 2012;12:425–36.

  6. 6.

    Kalluri R, Weinberg RA. The basics of epithelial-mesenchymal transition. J Clin Investig. 2009;119:1420–8.

  7. 7.

    Nieto MA, Huang RY, Jackson RA, Thiery JP. Emt: 2016. Cell. 2016;166:21–45.

  8. 8.

    Moreno-Bueno G, Portillo F, Cano A. Transcriptional regulation of cell polarity in EMT and cancer. Oncogene. 2008;27:6958–69.

  9. 9.

    Hotz B, Arndt M, Dullat S, Bhargava S, Buhr HJ, Hotz HG. Epithelial to mesenchymal transition: expression of the regulators snail, slug, and twist in pancreatic cancer. Clin Cancer Res. 2007;13:4769–76.

  10. 10.

    Stewart CJ, McCluggage WG. Epithelial-mesenchymal transition in carcinomas of the female genital tract. Histopathology. 2013;62:31–43.

  11. 11.

    Gonzalez DM, Medici D. Signaling mechanisms of the epithelial-mesenchymal transition. Sci Signal. 2014;7:re8.

  12. 12.

    Valenta T, Hausmann G, Basler K. The many faces and functions of beta-catenin. EMBO J. 2012;31:2714–36.

  13. 13.

    White BD, Chien AJ, Dawson DW. Dysregulation of Wnt/beta-catenin signaling in gastrointestinal cancers. Gastroenterology. 2012;142:219–32.

  14. 14.

    Thiery JP. Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002;2:442–54.

  15. 15.

    Schmalhofer O, Brabletz S, Brabletz T. E-cadherin, beta-catenin, and ZEB1 in malignant progression of cancer. Cancer Metastasis Rev. 2009;28:151–66.

  16. 16.

    Coso OA, Chiariello M, Yu JC, Teramoto H, Crespo P, Xu N, et al. The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signaling pathway. Cell. 1995;81:1137–46.

  17. 17.

    Faruqi TR, Gomez D, Bustelo XR, Bar-Sagi D, Reich NC. Rac1 mediates STAT3 activation by autocrine IL-6. Proc Natl Acad Sci USA. 2001;98:9014–9.

  18. 18.

    Gjoerup O, Lukas J, Bartek J, Willumsen BM. Rac and Cdc42 are potent stimulators of E2F-dependent transcription capable of promoting retinoblastoma susceptibility gene product hyperphosphorylation. J Biol Chem. 1998;273:18812–8.

  19. 19.

    Perona R, Montaner S, Saniger L, Sanchez-Perez I, Bravo R, Lacal JC. Activation of the nuclear factor-kappaB by Rho, CDC42, and Rac-1 proteins. Genes Dev. 1997;11:463–75.

  20. 20.

    D’Apolito M, Guarnieri V, Boncristiano M, Zelante L, Savoia A. Cloning of the murine non-muscle myosin heavy chain IIA gene ortholog of human MYH9 responsible for May-Hegglin, Sebastian, Fechtner, and Epstein syndromes. Gene. 2002;286:215–22.

  21. 21.

    Golomb E, Ma X, Jana SS, Preston YA, Kawamoto S, Shoham NG, et al. Identification and characterization of nonmuscle myosin II-C, a new member of the myosin II family. J Biol Chem. 2004;279:2800–8.

  22. 22.

    Simons M, Wang M, McBride OW, Kawamoto S, Yamakawa K, Gdula D, et al. Human nonmuscle myosin heavy chains are encoded by two genes located on different chromosomes. Circ Res. 1991;69:530–9.

  23. 23.

    Cote GP, Robinson EA, Appella E, Korn ED. Amino acid sequence of a segment of the Acanthamoeba myosin II heavy chain containing all three regulatory phosphorylation sites. J Biol Chem. 1984;259:12781–7.

  24. 24.

    Rayment I, Rypniewski WR, Schmidt-Base K, Smith R, Tomchick DR, Benning MM, et al. Three-dimensional structure of myosin subfragment-1: a molecular motor. Science. 1993;261:50–58.

  25. 25.

    Winkelmann DA, Almeda S, Vibert P, Cohen C. A new myosin fragment: visualization of the regulatory domain. Nature. 1984;307:758–60.

  26. 26.

    Choi OH, Park CS, Itoh K, Adelstein RS, Beaven MA. Cloning of the cDNA encoding rat myosin heavy chain-A and evidence for the absence of myosin heavy chain-B in cultured rat mast (RBL-2H3) cells. J Muscle Res Cell Motil. 1996;17:69–77.

  27. 27.

    Du M, Wang G, Ismail TM, Gross S, Fernig DG, Barraclough R, et al. S100P dissociates myosin IIA filaments and focal adhesion sites to reduce cell adhesion and enhance cell migration. J Biol Chem. 2012;287:15330–44.

  28. 28.

    Beach JR, Hussey GS, Miller TE, Chaudhury A, Patel P, Monslow J, et al. Myosin II isoform switching mediates invasiveness after TGF-beta-induced epithelial-mesenchymal transition. Proc Natl Acad Sci USA. 2011;108:17991–6.

  29. 29.

    Betapudi V, Licate LS, Egelhoff TT. Distinct roles of nonmuscle myosin II isoforms in the regulation of MDA-MB-231 breast cancer cell spreading and migration. Cancer Res. 2006;66:4725–33.

  30. 30.

    Jacobs K, Van Gele M, Forsyth R, Brochez L, Vanhoecke B, De Wever O, et al. P-cadherin counteracts myosin II-B function: implications in melanoma progression. Mol Cancer. 2010;9:255.

  31. 31.

    Xia H, Ng SS, Jiang S, Cheung WK, Sze J, Bian XW, et al. miR-200a-mediated downregulation of ZEB2 and CTNNB1 differentially inhibits nasopharyngeal carcinoma cell growth, migration and invasion. Biochem Biophys Res Commun. 2010;391:535–41.

  32. 32.

    Schramek D, Sendoel A, Segal JP, Beronja S, Heller E, Oristian D, et al. Direct in vivo RNAi screen unveils myosin IIa as a tumor suppressor of squamous cell carcinomas. Science. 2014;343:309–13.

  33. 33.

    Coaxum SD, Tiedeken J, Garrett-Mayer E, Myers J, Rosenzweig SA, Neskey DM. The tumor suppressor capability of p53 is dependent on non-muscle myosin IIA function in head and neck cancer. Oncotarget. 2017;8:22991–3007.

  34. 34.

    Kim JH, Adelstein RS. LPA(1) -induced migration requires nonmuscle myosin II light chain phosphorylation in breast cancer cells. J Cell Physiol. 2011;226:2881–93.

  35. 35.

    Dulyaninova NG, House RP, Betapudi V, Bresnick AR. Myosin-IIA heavy-chain phosphorylation regulates the motility of MDA-MB-231 carcinoma cells. Mol Biol Cell. 2007;18:3144–55.

  36. 36.

    Nakashima M, Adachi S, Yasuda I, Yamauchi T, Kawaguchi J, Hanamatsu T, et al. Inhibition of Rho-associated coiled-coil containing protein kinase enhances the activation of epidermal growth factor receptor in pancreatic cancer cells. Mol Cancer. 2011;10:79.

  37. 37.

    Newell-Litwa KA, Horwitz R, Lamers ML. Non-muscle myosin II in disease: mechanisms and therapeutic opportunities. Dis Model Mech. 2015;8:1495–515.

  38. 38.

    Li D, Zhou J, Wang L, Shin ME, Su P, Lei X, et al. Integrated biochemical and mechanical signals regulate multifaceted human embryonic stem cell functions. J Cell Biol. 2010;191:631–44.

  39. 39.

    Zhou P, Li B, Liu F, Zhang M, Wang Q, Liu Y, et al. The epithelial to mesenchymal transition (EMT) and cancer stem cells: implication for treatment resistance in pancreatic cancer. Mol Cancer. 2017;16:52.

  40. 40.

    Diala I, Wagner N, Magdinier F, Shkreli M, Sirakov M, Bauwens S, et al. Telomere protection and TRF2 expression are enhanced by the canonical Wnt signalling pathway. EMBO Rep. 2013;14:356–63.

  41. 41.

    Fagotto F. Looking beyond the Wnt pathway for the deep nature of beta-catenin. EMBO Rep. 2013;14:422–33.

  42. 42.

    Wang C, Fu SY, Wang MD, Yu WB, Cui QS, Wang HR, et al. Zinc finger protein X-linked promotes expansion of EpCAM+cancer stem-like cells in hepatocellular carcinoma. Mol Oncol. 2017;11:455–69.

  43. 43.

    Liu CC, Cai DL, Sun F, Wu ZH, Yue B, Zhao SL, et al. FERMT1 mediates epithelial-mesenchymal transition to promote colon cancer metastasis via modulation of beta-catenin transcriptional activity. Oncogene. 2017;36:1779–92.

  44. 44.

    Chew TL, Masaracchia RA, Goeckeler ZM, Wysolmerski RB. Phosphorylation of non-muscle myosin II regulatory light chain by p21-activated kinase (gamma-PAK). J Muscle Res Cell Motil. 1998;19:839–54.

  45. 45.

    Heasman SJ, Ridley AJ. Mammalian Rho GTPases: new insights into their functions from in vivo studies. Nat Rev Mol Cell Biol. 2008;9:690–701.

  46. 46.

    Vicente-Manzanares M, Ma X, Adelstein RS, Horwitz AR. Non-muscle myosin II takes centre stage in cell adhesion and migration. Nat Rev Mol Cell Biol. 2009;10:778–90.

  47. 47.

    Zeng Q, Lagunoff D, Masaracchia R, Goeckeler Z, Cote G, Wysolmerski R. Endothelial cell retraction is induced by PAK2 monophosphorylation of myosin II. J Cell Sci. 2000;113(Pt 3):471–82.

  48. 48.

    Arozarena I, Sanchez-Laorden B, Packer L, Hidalgo-Carcedo C, Hayward R, Viros A, et al. Oncogenic BRAF induces melanoma cell invasion by downregulating the cGMP-specific phosphodiesterase PDE5A. Cancer Cell. 2011;19:45–57.

  49. 49.

    Strohm C, Barancik T, Bruhl ML, Kilian SA, Schaper W. Inhibition of the ER-kinase cascade by PD98059 and UO126 counteracts ischemic preconditioning in pig myocardium. J Cardiovasc Pharmacol. 2000;36:218–29.

  50. 50.

    Cowen D, Troncoso P, Khoo VS, Zagars GK, von Eschenbach AC, Meistrich ML, et al. Ki-67 staining is an independent correlate of biochemical failure in prostate cancer treated with radiotherapy. Clin Cancer Res. 2002;8:1148–54.

  51. 51.

    Liu T, Ye Y, Zhang X, Zhu A, Yang Z, Fu Y, et al. Downregulation of nonmuscle myosin IIA expression inhibits migration and invasion of gastric cancer cells via the cJun Nterminal kinase signaling pathway. Mol Med Rep. 2016;13:1639–44.

  52. 52.

    Derycke L, Stove C, Vercoutter-Edouart AS, De Wever O, Dolle L, Colpaert N, et al. The role of non-muscle myosin IIA in aggregation and invasion of human MCF-7 breast cancer cells. Int J Dev Biol. 2011;55:835–40.

  53. 53.

    Liu D, Zhang L, Shen Z, Tan F, Hu Y, Yu J, et al. Clinicopathological significance of NMIIA overexpression in human gastric cancer. Int J Mol Sci. 2012;13:15291–304.

  54. 54.

    Hoelzle MK, Svitkina T. The cytoskeletal mechanisms of cell-cell junction formation in endothelial cells. Mol Biol Cell. 2012;23:310–23.

  55. 55.

    Ivanov AI, Samarin SN, Bachar M, Parkos CA, Nusrat A. Protein kinase C activation disrupts epithelial apical junctions via ROCK-II dependent stimulation of actomyosin contractility. BMC Cell Biol. 2009;10:36.

  56. 56.

    Liao Q, Li R, Zhou R, Pan Z, Xu L, Ding Y, et al. LIM kinase 1 interacts with myosin-9 and alpha-actinin-4 and promotes colorectal cancer progression. Br J Cancer. 2017;117:563–71.

  57. 57.

    Li C, Ma H, Wang Y, Cao Z, Graves-Deal R, Powell AE, et al. Excess PLAC8 promotes an unconventional ERK2-dependent EMT in colon cancer. J Clin Investig. 2014;124:2172–87.

  58. 58.

    Shin S, Dimitri CA, Yoon SO, Dowdle W, Blenis J. ERK2 but not ERK1 induces epithelial-to-mesenchymal transformation via DEF motif-dependent signaling events. Mol Cell. 2010;38:114–27.

  59. 59.

    Smith BN, Burton LJ, Henderson V, Randle DD, Morton DJ, Smith BA, et al. Snail promotes epithelial mesenchymal transition in breast cancer cells in part via activation of nuclear ERK2. PLoS ONE. 2014;9:e104987.

  60. 60.

    Ning BF, Ding J, Yin C, Zhong W, Wu K, Zeng X, et al. Hepatocyte nuclear factor 4 alpha suppresses the development of hepatocellular carcinoma. Cancer Res. 2010;70:7640–51.

  61. 61.

    Thaker PH, Deavers M, Celestino J, Thornton A, Fletcher MS, Landen CN, et al. EphA2 expression is associated with aggressive features in ovarian carcinoma. Clin Cancer Res. 2004;10:5145–50.

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Funding

This work was supported by National Natural Science Foundation of China (grant 81370600, DL); The Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning (TP2015022, DL); Shanghai Pujiang Program (15PJ1404800, DL); Innovation Program of Shanghai Municipal Education Commission (15ZZ056, DL); and Innovation Program for Ph.D. students in Shanghai Jiaotong University School of Medicine (BXJ201731).

Authors’ contributions

Study design and concept: LD, ZPT. Data acquisition: ZPT, LYY, LB, LYH. Data analysis and interpretation: ZPT. Manuscript preparation: ZPT, LB, LD. Manuscript review: LD, YY, LB. All authors read and approved the final manuscript.

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Author notes

  1. These authors have contributed equally: Pingting Zhou, Yanyan Li and Bo Li

Affiliations

  1. Department of Radiation Oncology, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

    • Pingting Zhou
    • , Yanyan Li
    • , Meichao Zhang
    • , Yuan Yao
    •  & Dong Li
  2. Department of Orthopedic Oncology, Changzheng Hospital, Second Military Medical University, Shanghai, China

    • Bo Li
  3. Department of Chemotherapy, Nanjing Medical University Affiliated Cancer Hospital, Cancer Institute of Jiangsu Province, Nanjing, Jiangsu, China

    • Yuanhua Liu

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

All procedures of human and mouse experiments were approved by Ethics Committee of Shanghai Ninth People’s Hospital affiliated to Shanghai JiaoTong University, School of Medicine.

Corresponding authors

Correspondence to Yuanhua Liu or Yuan Yao or Dong Li.

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https://doi.org/10.1038/s41388-019-0806-6