Advertisement

Folic acid prevents the progesterone-promoted proliferation and migration in breast cancer cell lines

  • Hui-Chen Wang
  • Yen-Nien Huo
  • Wen-Sen LeeEmail author
Original Contribution
  • 88 Downloads

Abstract

Purpose

We previously demonstrated that progesterone (P4) interacted with folic acid (FA) and abolished the FA-reduced endothelial cell proliferation and migration. These findings led us to investigate whether FA can interfere with the P4-promoted breast cancer cell proliferation and migration.

Methods

We conducted MTT and wound healing assay to evaluate cell proliferation and migration, respectively. Western blot analysis and immunoprecipitation were performed to examine the protein expression and protein–protein interaction, respectively.

Results

We demonstrated that P4 promoted proliferation and migration of breast cancer cell lines (T47D, MCF-7, BT474, and BT483). However, co-treatment with P4 and FA together abolished these promotion effects. Treatment with P4 alone increased the formation of PR-cSrc complex and the phosphorylation of cSrc at tyrosine 416 (Tyr416). However, co-treatment with P4 and FA together increased the formations of cSrc-p140Cap, cSrc-Csk, and cSrc-p-Csk complex, and the phosphorylation of cSrc at tyrosine 527 (Tyr527). Co-treatment with P4 and FA together also abolished the activation of cSrc-mediated signaling pathways involved in the P4-promoted breast cancer cell proliferation and migration.

Conclusions

Co-treatment with FA and P4 together abolished the P4-promoted breast cancer cell proliferation and migration through decreasing the formation of PR-cSrc complex and increasing the formations of cSrc-p140Cap and cSrc-Csk complex, subsequently activating Csk, which in turn suppressed the phosphorylation of cSrc at Tyr416 and increased the phosphorylation of cSrc at Tyr527, hence inactivating the cSrc-mediated signaling pathways. The findings from this study might provide a new strategy for preventing the P4-promoted breast cancer progress.

Keywords

Csk cSrc p140Cap p-cSrcY416 p-cSrcY527 

Notes

Acknowledgements

This work was supported by the research grant from the Ministry of Science and Technology, R.O.C. (MOST 107-2320-B-038 -051 -MY3).

Compliance with ethical standards

Conflict of interests

The authors have not conflict of interest.

References

  1. 1.
    MacMahon B, Cole BP, Brown J (1973) Etiology of human breast cancer: a review. J Natl Cancer Inst 50(1):21–42.  https://doi.org/10.1093/jnci/50.1.21 CrossRefGoogle Scholar
  2. 2.
    Dao TL (1981) The role of ovarian steroid hormones in mammary carcinogenesis. In: Pike MC, Siiteri PK, Welsch CW (eds) Hormones and breast cancer. Banbury report no. 8. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 281–289Google Scholar
  3. 3.
    Pike MC, Spicer DV, Dahmoush L, Press MF (1993) Estrogens, progestogens, normal breast cell proliferation, and breast cancer risk. Epidemiol Rev 15(1):7–35.  https://doi.org/10.1093/oxfordjournals.epirev.a036102 CrossRefGoogle Scholar
  4. 4.
    Key T, Appleby P, Barnes I, Reeves G, Endogenous Hormones and Breast Cancer Collaborative Group (2002) Endogenous sex hormones and breast cancer in postmenopausal women: reanalysis of nine prospective studies. J Natl Cancer Inst 94(8):606–616.  https://doi.org/10.1093/jnci/94.8.606 CrossRefGoogle Scholar
  5. 5.
    Beral V, Million Women Study Collaborators (2003) Breast cancer and hormone-replacement therapy in the Million Women Study. Lancet 362(9382):419–427.  https://doi.org/10.1016/S0140-6736(03)14065-2 CrossRefGoogle Scholar
  6. 6.
    Anderson GL, Limacher M, Assaf AR, Bassford T, Beresford SA, Black H, Bonds D, Brunner R, Brzyski R, Caan B et al (2004) Effects of conjugated equine estrogen in postmenopausal women with hysterectomy: the Women’s Health Initiative randomized controlled trial. JAMA 291(14):1701–1712.  https://doi.org/10.1210/me.2006-0337 CrossRefGoogle Scholar
  7. 7.
    Travis RC, Key TJ (2003) Oestrogen exposure and breast cancer risk. Breast Cancer Res 5(5):239–247.  https://doi.org/10.1001/jama.291.14.1701 CrossRefGoogle Scholar
  8. 8.
    Boonyaratanakornkit V, McGowan E, Sherman L, Mancini MA, Cheskis BJ, Edwards DP (2007) The role of extranuclear signaling actions of progesterone receptor in mediating progesterone regulation of gene expression and the cell cycle. Mol Endocrinol 21(2):359–375.  https://doi.org/10.1210/me.2006-0337 CrossRefGoogle Scholar
  9. 9.
    Skildum A, Faivre E, Lange CA (2005) Progesterone receptors induce cell cycle progression via activation of mitogen-activated protein kinases. Mol Endocrinol 19(2):327–339.  https://doi.org/10.1210/me.2004-0306 CrossRefGoogle Scholar
  10. 10.
    Saitoh M, Ohmichi M, Takahashi K, Kawagoe J, Ohta T, Doshida M, Takahashi T, Igarashi H, Mori-Abe A, Du B, Tsutsumi S, Kurachi H (2005) Medroxyprogesterone acetate induces cell proliferation through up-regulation of cyclin D1 expression via phosphatidylinositol 3-kinase/Akt/nuclear factor-kappaB cascade in human breast cancer cells. Endocrinology 146(11):4917–4925.  https://doi.org/10.1210/en.2004-1535 CrossRefGoogle Scholar
  11. 11.
    Fu XD, Giretti MS, Baldacci C, Garibaldi S, Flamini M, Sanchez AM, Gadducci A, Genazzani AR, Simoncini T (2008) Extra-nuclear signaling of progesterone receptor to breast cancer cell movement and invasion through the actin cytoskeleton. PLoS One 3(7):e2790.  https://doi.org/10.1371/journal.pone.0002790 CrossRefGoogle Scholar
  12. 12.
    Wang HC, Lee WS (2016) Molecular mechanisms underlying progesterone-enhanced breast cancer cell migration. Sci Rep 6:31509.  https://doi.org/10.1038/srep31509 CrossRefGoogle Scholar
  13. 13.
    Wang HC, Lee WS (2018) Molecular mechanisms underlying progesterone-induced cytoplasmic retention of p27 in breast cancer cells. J Steroid Biochem Mol Biol 183:202–209.  https://doi.org/10.1016/j.jsbmb.2014.12.002 CrossRefGoogle Scholar
  14. 14.
    Hsu SP, Lee WS (2011) Progesterone receptor activation of extranuclear signaling pathways in regulating p53 expression in vascular endothelial cells. Mol Endocrinol 25(3):421–432.  https://doi.org/10.1210/me.2010-0424 CrossRefGoogle Scholar
  15. 15.
    Lin SY, Lee WR, Su YF, Lee WS (2012) Folic acid inhibits endothelial cell proliferation through activating the cSrc/ERK 2/NF-kB/p53 pathway mediated by folic acid receptor. Angiogenesis 15(4):671–683.  https://doi.org/10.1007/s10456-012-9289-6 CrossRefGoogle Scholar
  16. 16.
    Hou TC, Lin JJ, Wen HC, Chen LC, Hsu SP, Lee WS (2013) Folic acid inhibits endothelial cell migration through inhibiting the RhoA activity mediated by activating the folic acid receptor/c-SRC/p190RhoGAP-signaling pathway. Biochem Pharmacol 85(3):376–384.  https://doi.org/10.1016/j.bcp.2012.11.011 CrossRefGoogle Scholar
  17. 17.
    Lee TS, Lin JJ, Huo YN, Lee WS (2015) Progesterone inhibits endothelial cell migration through suppression of the rho activity mediated by cSrc activation. J Cell Biochem 116(7):1411–1418.  https://doi.org/10.1002/jcb.25101 CrossRefGoogle Scholar
  18. 18.
    Lee WS, Lu YC, Kuo CT, Chen CT, Tang PH (2018) Effects of female sex hormones on folic acid–induced anti-angiogenesis. Acta Physiol 222(4):e13001.  https://doi.org/10.1111/apha.13001 CrossRefGoogle Scholar
  19. 19.
    Wang HC, Lee WS (2014) Progesterone-induced migration inhibition in male rat aortic smooth muscle cells through the cSrc/AKT/ERK 2/p38 pathway-mediated up-regulation of p27. Endocrinology 155(4):1428–1435.  https://doi.org/10.1210/en.2013-1838 CrossRefGoogle Scholar
  20. 20.
    Wang HC, Hsu SP, Lee WS (2015) Extra-nuclear signaling pathway involved in progesterone-induced up-regulations of p21cip1 and p27kip1 in rat aortic smooth muscle cells. PLoS One 10(5):e0125903.  https://doi.org/10.1371/journal.pone.0125903 CrossRefGoogle Scholar
  21. 21.
    Wang HC, Lee WS (2014) Progesterone induces RhoA inactivation in male rat aortic smooth muscle cells through up-regulation of p27kip1. Endocrinology 155(11):4473–4482.  https://doi.org/10.1210/en.2014-1344 CrossRefGoogle Scholar
  22. 22.
    Wen HC, Huo YN, Chou CM, Lee WS (2018) PMA inhibits endothelial cell migration through activating the PKC-δ/Syk/NF-κB-mediated upregulation of Thy-1. Sci Rep 8(1):16247.  https://doi.org/10.1038/s41598-018-34548-8 CrossRefGoogle Scholar
  23. 23.
    Lee WS, Harder JA, Yoshizumi M, Lee ME, Haber E (1997) Progesterone inhibits arterial smooth muscle cell proliferation. Nat Med 3(9):1005–1008.  https://doi.org/10.1038/nm0997-1005 CrossRefGoogle Scholar
  24. 24.
    Daly LE, Kirke PN, Molloy A, Weir DG, Scott JM (1995) Folate levels and neural tube defects. Implications for prevention. JAMA 274(21):1698–1702.  https://doi.org/10.1001/jama.1995.03530210052030 CrossRefGoogle Scholar
  25. 25.
    Chong YP, Ia KK, Mulhern TD, Cheng HC (2005) Endogenous and synthetic inhibitors of the Src-family protein tyrosine kinases. Biochim Biophys Acta 1754(1–2):210–220.  https://doi.org/10.1016/j.bbapap.2005.07.027 CrossRefGoogle Scholar
  26. 26.
    Di Stefano P, Damiano L, Cabodi S, Aramu S, Tordella L, Parduroux A, Pica R, Cavallo F, Forni G, Silengo L, Tarone G, Turco E, Defilippi P (2007) p140Cap protein suppresses tumour cell properties, regulating Csk and Src kinase activity. EMBO J 26(12):2843–2855.  https://doi.org/10.1038/sj.emboj.7601724 CrossRefGoogle Scholar
  27. 27.
    Kuo CT, Lee WS (2016) Progesterone receptor activation is required for folic acid-induced anti-proliferation in colorectal cancer cell lines. Cancer Lett 378(2):104–110.  https://doi.org/10.1016/j.canlet.2016.05.019 CrossRefGoogle Scholar
  28. 28.
    Collett MS, Purchio AF, Erikson RL (1980) Avian sarcoma virus-transforming protein, pp60src shows protein kinase activity specific for tyrosine. Nature 285(5761):167–169.  https://doi.org/10.1038/285167a0 CrossRefGoogle Scholar
  29. 29.
    Hunter T, Sefton BM, Beemon K, Eckhart W (1980) Evidence that the phosphorylation of tyrosine is essential for cellular transformation by Rous sarcoma virus. Cell 20(3):807–816.  https://doi.org/10.1016/0092-8674(80)90327-X CrossRefGoogle Scholar
  30. 30.
    Summy JM, Gallick GE (2003) Src family kinases in tumor progression and metastasis. Cancer Metastasis Rev 22(4):337–358.  https://doi.org/10.1023/a:1023772912750 CrossRefGoogle Scholar
  31. 31.
    Irby RB, Yeatman TJ (2000) Role of Src expression and activation in human cancer. Oncogene 19(49):5636–5642.  https://doi.org/10.1038/sj.onc.1203912 CrossRefGoogle Scholar
  32. 32.
    Mao W, Irby R, Coppola D, Fu L, Wloch M, Turner J, Yu H, Garcia R, Jove R, Yeatman TJ (1997) Activation of c-Src by receptor tyrosine kinases in human colon cancer cells with high metastatic potential. Oncogene 15(25):3083–3090.  https://doi.org/10.1038/sj.onc.1201496 CrossRefGoogle Scholar
  33. 33.
    Hamaguchi M, Matsuyoshi N, Ohnishi Y, Gotoh B, Takeichi M, Nagai Y (1993) p60v-src causes tyrosine phosphorylation and inactivation of the N-cadherin-catenin cell adhesion system. EMBO J 12(1):307–314.  https://doi.org/10.1002/j.1460-2075.1993.tb05658.x CrossRefGoogle Scholar
  34. 34.
    Irby RB, Mao W, Coppola D, Kang J, Loubeau JM, Trudeau W (1999) Activating SRC mutation in a subset of advanced human colon cancers. Nat Genet 21(2):187–190.  https://doi.org/10.1038/5971 CrossRefGoogle Scholar
  35. 35.
    Cowan-Jacob SW, Fendrich G, Manley PW, Jahnke W, Fabbro D, Liebetanz J, Meyer T (2005) The crystal structure of a c-Src complex in an active conformation suggests possible steps in c-Src activation. Structure 13(6):861–871.  https://doi.org/10.1016/j.str.2005.03.012 CrossRefGoogle Scholar
  36. 36.
    Xu Y, Singer MA, Lindquist S (1999) Maturation of the tyrosine kinase c-src as a kinase and as a substrate depends on the molecular chaperone Hsp90. Proc Natl Acad Sci USA 96(1):109–114.  https://doi.org/10.1073/pnas.96.1.109 CrossRefGoogle Scholar
  37. 37.
    Okada M (2012) Regulation of the SRC family kinases by Csk. Int J Biol Sci 8:1385–1397.  https://doi.org/10.7150/ijbs.5141 CrossRefGoogle Scholar
  38. 38.
    Somani AK, Bignon JS, Mills GB, Siminovitch KA, Branch DR (1997) Src kinase activity is regulated by the SHP-1 protein-tyrosine phosphatase. J Biol Chem 272(34):21113–21119.  https://doi.org/10.1074/jbc.272.34.21113 CrossRefGoogle Scholar
  39. 39.
    Boonyaratanakornkit V, Scott MP, Ribon V, Sherman L, Anderson SM, Maller JL, Miller WT, Edwards DP (2001) Progesterone receptor contains a proline-rich motif that directly interacts with SH3 domains and activates c-Src family tyrosine kinases. Mol Cell 8(2):269–280.  https://doi.org/10.1016/S1097-2765(01)00304-5 CrossRefGoogle Scholar
  40. 40.
    Kastner P, Krust A, Turcotte B, Stropp U, Tora L, Gronemeyer H, Chambon P (1990) Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. EMBO J 9(5):1603–1614.  https://doi.org/10.1002/j.1460-2075.1990.tb08280.x CrossRefGoogle Scholar
  41. 41.
    Cheung J, Smith DF (2000) Molecular chaperone interactions with steroid receptors: an update. Mol Endocrinol 14(7):939–946.  https://doi.org/10.1210/mend.14.7.0489 CrossRefGoogle Scholar
  42. 42.
    Conneely OM, Mulac-Jericevic B, Lydon JP (2003) Progesterone-dependent regulation of female reproductive activity by two distinct progesterone receptor isoforms. Steroids 68(10–13):771–778.  https://doi.org/10.1016/S0039-128X(03)00126-0 CrossRefGoogle Scholar
  43. 43.
    He B, Kemppainen JA, Wilson EM (2000) FXXLF and WXXLF sequences mediate the NH2-terminal interaction with the ligand binding domain of the androgen receptor. J Biol Chem 275(30):22986–22994.  https://doi.org/10.1074/jbc.M002807200 CrossRefGoogle Scholar
  44. 44.
    El-Ashry D, Onate SA, Nordeen SK, Edwards DP (1989) Human progesterone receptor complexed with the antagonist RU 486 binds to hormone response elements in a structurally altered form. Mol Endocrinol 3(10):1545–1558.  https://doi.org/10.1210/mend-3-10-1545 CrossRefGoogle Scholar
  45. 45.
    Gellersen B, Fernandes MS, Brosens JJ (2009) Non-genomic progesterone actions in female reproduction. Hum Reprod Update 15(1):119–138.  https://doi.org/10.1093/humupd/dmn044 CrossRefGoogle Scholar
  46. 46.
    Kay BK, Williamson MP, Sudol M (2000) The importance of being proline: the interaction of proline-rich motifs in signaling proteins with their cognate domains. FASEB J 14(2):231–241.  https://doi.org/10.1096/fasebj.14.2.231 CrossRefGoogle Scholar
  47. 47.
    Nada S, Okada M, MacAuley A, Cooper JA, Nakagawa H (1991) Cloning of a complementary DNA for a protein-tyrosine kinase that specifically phosphorylates a negative regulatory site of p60c-src. Nature 351(6321):69–72.  https://doi.org/10.1038/351069a0 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Graduate Institute of Medical Sciences, College of MedicineTaipei Medical UniversityTaipeiTaiwan
  2. 2.Department of Physiology, School of Medicine, College of MedicineTaipei Medical UniversityTaipeiTaiwan
  3. 3.Cancer Research CenterTaipei Medical University HospitalTaipeiTaiwan
  4. 4.Cell Physiology and Molecular Image Research Center, Wan Fang HospitalTaipei Medical UniversityTaipeiTaiwan

Personalised recommendations