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04.03.2019 | Original Article

Breast cancer cell motility is promoted by 14-3-3γ

verfasst von: Emiko Hiraoka, Takahiro Mimae, Masaoki Ito, Takayuki Kadoya, Yoshihiro Miyata, Akihiko Ito, Morihito Okada

Erschienen in: Breast Cancer | Ausgabe 5/2019

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Abstract

Purpose

Pseudopodia are actin-rich ventral protrusions associated with cell motility and cancer cell invasion. We previously applied our established method of using excimer laser cell etching to isolate pseudopodial proteins from MDA-MB-231 breast cancer cells. We later identified 14-3-3γ as an oncogenic molecule among 46 candidate proteins that are specific to pseudopodia. The present study aimed to determine the function of 14-3-3γ in the motility of breast cancer cells.

Methods

MDA-MB-231 cells were cultured on 3-µm porous membranes and double stained to localize 14-3-3γ and phalloidin in pseudopodia using confocal imaging. We assessed pseudopodia numbers and length, as well as migration and wound healing in MDA-MB-231 cells with knockdown and forced expression of 14-3-3γ to determine 14-3-3γ involvement in cell motility. We also immunohistochemically analyzed 14-3-3γ in human breast cancer tissues with high-grade lymphatic invasion.

Results

We specifically located 14-3-3γ in pseudopodia of MDA-MB-231 cells. Knockdown and forced expression of 14-3-3γ, respectively, decreased and increased pseudopodial formation and elongation. Migration and wound healing assays also showed that 14-3-3γ knockdown and forced expression, respectively, decreased and increased the number of underside cells and acellular areas in MDA-MB-231 breast cancer cells. More 14-3-3γ was expressed in sites of lymphatic invasion, than in the center and periphery of human breast cancer tissues.

Conclusion

The role of 14-3-3γ in breast cancer invasiveness might be to promote cell motility. Inhibition of 14-3-3γ could, therefore, become a novel target of therapy to prevent invasion and metastasis in patients with breast cancers expressing 14-3-3γ.

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Guirguis R, Margulies I, Taraboletti G, Schiffmann E, Liotta L. Cytokine-induced pseudopodial protrusion is coupled to tumour cell migration. Nature. 1987;329:261–3.CrossRefPubMed

Lauffenburger DA, Horwitz AF. Cell migration: a physically integrated molecular process. Cell. 1996;84(3):359–69.CrossRefPubMed

Bravo-Cordero JJ, Hodgson L, Condeelis J. Directed cell invasion and migration during metastasis. Curr Opin Cell Biol. 2012;24(2):277–83.CrossRefPubMed

Ito A, Mimae T, Yamamoto YS, Hagiyama M, Nakanishi J, Ito M, Hosokawa Y, Okada M, Murakami Y, Kondo T. Novel application for pseudopodia proteomics using excimer laser ablation and two-dimensional difference gel electrophoresis. Lab Investig. 2012;92(9):1374–85.CrossRefPubMed

Mimae T, Ito A. New challenges in pseudopodial proteomics by a laser-assisted cell etching technique. Biochim Biophys Acta. 2015;1854(6):538–46.CrossRefPubMed

Wang X, Liu F, Qin X, Huang T, Huang B, Zhang Y, Jiang B. Expression of Rab1A is upregulate in human lung cancer and associated with tumor size and T stage. Aging. 2016;8(11):2790–8.CrossRefPubMedPubMedCentral

Wang ZK, Cheng ZW, Chen SJ, Zhu XG, Gu YP, Yang XD, Sun L, Liu WT, Zhang YJ, Yuan JF, Tian KJ, Yao YZ, He SB. Aberrant expression of Rab1A and its prognostic significance in human colorectal cancer. Eur Rev Med Pharmacol Sci. 2018;22(14):4509–17.PubMed

Xu B, Huang C, Yang X, Li X, Li L, Ding Y. Significance and prognostic role of human epidermal growth factor receptor 2 and RAB1A expression in gastric cancer. Oncol Lett. 2018;15(4):5185–92.PubMedPubMedCentral

Xu H, Qian M, Zhao B, Wu C, Maskey N, Song H, Li D, Song J, Hua K, Fang L. Inhibition of RAB1A suppresses epithelial-mesenchymal transition and proliferation of triple-negative breast cancer cells. Oncol Rep. 2017;37(3):1619–26.CrossRefPubMed

10.

Ito M, Hagiyama M, Mimae T, Inoue T, Kato T, Yoneshige A, Nakanishi J, Kondo T, Okada M, Ito A. α-Parvin, a pseudopodial constituent, promotes cell motility and is associated with lymph node metastasis of lobular breast carcinoma. Breast Cancer Res Treat. 2014;144(1):59–69.CrossRefPubMed

11.

Muslin AJ, Tanner JW, Allen PM, Shaw AS. Interaction of 14-3-3 with signaling proteins is mediated by the recognition of phosphoserine. Cell. 1996;84(6):889–97.CrossRefPubMed

12.

Fu H, Subramanian RR, Masters SC. 14-3-3 proteins: structure, function, and regulation. Annu Rev Pharmacol Toxicol. 2000;40:617–47.CrossRefPubMed

13.

Dougherty MK, Morrison DK. Unlocking the code of 14-3-3. J Cell Sci. 2004;117(Pt 10):1875–84.CrossRefPubMed

14.

Aitken A, Collinge DB, van Heusden BP, Isobe T, Roseboom PH, Rosenfeld G, Soll J. 14-3-3 proteins: a highly conserved, widespread family of eukaryotic proteins. Trends Biochem Sci. 1992;17(12):498–501.CrossRefPubMed

15.

Mackintosh C. Dynamic interactions between 14-3-3 proteins and phosphoproteins regulated diverse cellular processes. Biochem J. 2004;381(Pt 2):329–42.CrossRefPubMedPubMedCentral

16.

Wu YJ, Jan YJ, Ko BS, Liang SM, Liou JY. Involvement of 14-3-3 proteins in regulating tumor progression of hepatocellular carcinoma. Cancers (Basal). 2015;7:1022–36.CrossRef

17.

Liu TA, Jan YJ, Ko BS, Chen SC, Liang SM, Hung YL, Hsu C, Shen TL, Lee YM, Chen PF, Wang J, Shyue SK, Liou JY. Increased expression of 14-3-3β promotes tumor progression and predicts extrahepatic metastasis and worse survival in hepatocellular carcinoma. Am J Pathol. 2011;179(6):2698–708.CrossRefPubMedPubMedCentral

18.

Sugiyama A, Miyagi Y, Komiya Y, Kurabe N, Kitanaka C, Kato N, Nagashima Y, Kuchino Y, Tashiro F. Forced expression of antisense 14-3-3β RNA suppresses tumor cell growth in vitro and in vivo. Carcinogenesis. 2003;24:1549–59.CrossRefPubMed

19.

Choi JE, Hur W, Jung CK, Piao LS, Lyoo K, Hong SW, Kim SW, Yoon HY, Yoon SK. Silencing of 14-3-3ζ over-expression in hepatocellular carcinoma inhibits tumor growth and enhances chemosensitivity to cis-diamminedichloroplatinum. Cancer Lett. 2011;303(2):99–107.CrossRefPubMed

20.

Liu TA, Jan YJ, Ko BS, Liang SM, Chen SC, Wang J, Hsu C, Wu YM, Lion JY. 14-3-3ε overexpression contributes to epithelial-mesenchymal transition of hepatocellular carcinoma. PLoS ONE. 2013;8(3):e57968.CrossRefPubMedPubMedCentral

21.

Ko BS, Jan YJ, Chang TC, Liang SM, Chen SC, Liu TA, Wu YM, Wang J, Liou JY. Upregulation of focal adhesion kinase by 14-3-3ε via NFκB activation in hepatocellular carcinoma. Anticancer Agents Med Chem. 2013;13:555–62.CrossRefPubMed

22.

Liu CC, Jan YJ, Ko BS, Liang SM, Chen SC, Lee YM, Liu TA, Chang TC, Wang J, Shyue SK, Sung LY, Liou JY. 14-3-3σ induces heat shock protein 70 expression in hepatocellular carcinoma. BMC Cancer. 2014;14:425. https://doi.org/10.1186/1471-2407-14-425.CrossRefPubMedPubMedCentral

23.

Lee IN, Chen CH, Sheu JC, Lee HS, Huang GT, Yu CY, Lu FJ, Chow LP. Identification of human hepatocellular carcinoma-related biomarkers by two-dimensional difference gel electrophoresis and mass spectrometry. J Proteome Res. 2005;4:2062–9.CrossRefPubMed

24.

Raungrut P, Wongkotsila A, Lirdprapamomgkol K, Svasti J, Geater SL, Phukaoloun M, Suwiwat S, Thonqsuksai P. Prognostic significance of 14-3-3γ overexpression in advanced non-small cell lung cancer. Asian Pac J Cancer Prev. 2014;15(8):3513–8.CrossRefPubMed

25.

Teo Z, Sng MK, Lim MMK, Li Y, Li L, Phua T, Lee JYH, Tan ZW, Zhu P, Tan NS. Elevation of adenylate energy charge by angiopoietin-like 4 enhances epithelial-mesenchymal transition by inducing 14-3-3γ expression. Oncogene. 2017;36(46):6408–19.CrossRefPubMedPubMedCentral

26.

Song Y, Yang Z, Ke Z, Yao Y, Hu X, Sun Y, Li H, Yin J, Zeng C. Expression of 14-3-3γ in patients with breast cancer: correlation with clinicopathological features and prognosis. Cancer Epidemiol. 2012;36(6):533–6.CrossRefPubMed

27.

Ajjappala BS, Kim YS, Kim MS, Lee KY, Ki HY, Cha DH, Baek KH. 14-3-3γ is stimulated by il-3 and promotes cell proliferation. J Immunol. 2009;182(2):1050–60.CrossRefPubMed

28.

Ko BS, Lai IR, Chang TC, Liu TA, Chen SC, Wang J, Liou JY. Involvement of 14-3-3γ overexpression in extrahepatic metastasis of hepatocellular carcinoma. Hum Pathol. 2011;42(1):129–35.CrossRefPubMed

29.

Wang P, Deng Y, Fu X. MiR-509-5p suppresses the proliferation, migration, and invasion of non-small cell lung cancer by targeting YWHAG. Biochem Biophys Res Commun. 2017;482(4):935–41.CrossRefPubMed

30.

Raungrut P, Wongkotsila A, Champoochana N, Lirdprapamomgkol K, Svasti J, Svasti J, Thonqsuksai P. Knockdown of 14-3-3γ suppresses epithelial-mesenchymal transition and reduces metastatic potential of human non-small cell lung cancer cells. Anticancer Res. 2018;38(6):3507–14.CrossRefPubMed

31.

Kanda Y. Investigation of the freely-available easy-to-use software “EZR” (Easy R) for medical statistics. Bone Marrow Transpl. 2013;48(3):452–8.CrossRef

32.

Perdigão-Henriques R, Petrocca F, Altschuler G, Thomas MP, Le MT, Tan SM, Hide W, Lieberman J. miR-200 promotes the mesenchymal to epithelial transition by suppressing multiple members of the Zeb2 and Snail1 transcriptional repressor complexes. Oncogene. 2016;35(2):158–72.CrossRefPubMed

Titel: Breast cancer cell motility is promoted by 14-3-3γ
verfasst von: Emiko Hiraoka
Takahiro Mimae
Masaoki Ito
Takayuki Kadoya
Yoshihiro Miyata
Akihiko Ito
Morihito Okada
Publikationsdatum: 04.03.2019
Verlag: Springer Japan
Erschienen in: Breast Cancer / Ausgabe 5/2019
Print ISSN: 1340-6868
Elektronische ISSN: 1880-4233
DOI: https://doi.org/10.1007/s12282-019-00957-4

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Breast cancer cell motility is promoted by 14-3-3γ