Tumor Biology

, Volume 36, Issue 4, pp 2831–2844 | Cite as

LASS2/TMSG1 inhibits growth and invasion of breast cancer cell in vitro through regulation of vacuolar ATPase activity

  • Fang Mei
  • Jiangfeng You
  • Beiying Liu
  • Mengxue Zhang
  • Jiangying Liu
  • Bo Zhang
  • Fei Pei
Research Article

Abstract

Homo sapiens longevity assurance homologue 2 of yeast LAG1 (LASS2)/tumor metastasis suppressor gene 1 (TMSG1) was a novel tumor metastasis-related gene identified using messenger RNA differential display from non-metastatic human prostate cancer cell variants. The mechanism of LASS2/TMSG1 inhibiting tumor invasion metastasis in breast cancer cells had not been well investigated. In the present study, a full length of 1.2 kb LASS2/TMSG1 complementary DNA (cDNA) coding for a protein of 380 amino acids was cloned. PcDNA3 eukaryotic expression plasmids of LASS2/TMSG1 were constructed and transfected into human breast cancer cell line MCF-7 by lipofectin transfection method. And, the biological effects were observed comparing with control groups. As the result, LASS2/TMSG1 inhibited cell growth in vitro by increasing apoptosis and changing cell cycle distribution. Furthermore, the vacuolar ATPase (V-ATPase) activity and extracellular hydrogen ion concentration were significantly decreased and the activity of secreted matrix metalloproteinase-2 (MMP-2) was downregulated in MCF-7 cells overexpressing LASS2/TMSG1 compared with the controls. Therefore, LASS2/TMSG1 may inhibit growth and invasion of breast cancer cell in vitro through decreasing V-ATPase activity and extracellular hydrogen ion concentration and inactivating secreted MMP-2. The findings provided the evidence that the LASS2/TMSG1 gene had tumor growth and invasion suppressor function in human breast cancer cell and may provide a promising target for cancer metastasis diagnosis and therapy.

Keywords

LASS2/TMSG1 Breast cancer Vacuolar ATPase MMP-2 

Notes

Acknowledgments

The article was supported by the National Sciences Foundation of China (30971142). Thanks to Beiying Liu for data analysis and figure modification.

References

  1. 1.
    Erez-Roman R, Pienik R, Futerman AH. Increased ceramide synthase 2 and 6 mRNA levels in breast cancer tissues and correlation with sphingosine kinase expression. Biochem Biophys Res Commun. 2010;391:219–23.CrossRefPubMedGoogle Scholar
  2. 2.
    Fan S, Niu Y, Tan N, Wu Z, Wang Y, You H, et al. LASS2 enhances chemosensitivity of breast cancer by counteracting acidic tumor microenvironment through inhibiting activity of V-ATPase proton pump. Oncogene. 2012;32(13):1682–90.CrossRefPubMedGoogle Scholar
  3. 3.
    Fasciglione GF, Marini S, D’Alessio S, Politi V, Coletta M. pH- and temperature-dependence of functional modulation in metalloproteinases: a comparison between neutrophil collagenase and gelatinases A and B. Biophys J. 2000;79(4):2138–49.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Forgac M. Structure and properties of the vacuolar (H+)-ATPases. J Biol Chem. 1999;274(19):12951–4.CrossRefPubMedGoogle Scholar
  5. 5.
    Jiang P, Enomoto A, Kanahashi M. Cell biology of the movement of breast cancer cells: intracellular signalling and the actin cytoskeleton. Cancer Lett. 2009;284(2):122–30.CrossRefPubMedGoogle Scholar
  6. 6.
    Johnson LL, Pavlovsky AG, Johnson AR, Janowicz JA, Man CF, Ortwine DF, et al. A rationalization of the acidic pH dependence for stromelysin-1 (matrix metalloproteinase-3) catalysis and inhibition. J Biol Chem. 2000;275(15):11026–33.CrossRefPubMedGoogle Scholar
  7. 7.
    Kraveka JM, Li L, Szulc ZM, Bielawski J, Ogretmen B, Hannun YA, et al. Involvement of dihydroceramide desaturase in cell cycle progression in human neuroblastoma cells. J Biol Chem. 2007;282:16718–28.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Laviad EL, Albee L, Pankova-Kholmyansky I, Epstein S, Park H, Merrill AH, et al. Characterization of ceramide synthase 2-tissue distribution, substrate specificity, and inhibition by sphingosine 1-phosphate. J Biol Chem. 2008;283(9):5677–84.CrossRefPubMedGoogle Scholar
  9. 9.
    Liu Y, Zheng J, Fang W, You J, Wang J, Cui X, et al. Isolation and characterization of human prostate cancer cell subclones with different metastatic potential. Zhonghua Bing Li Xue Za Zhi (Chinese). 1999;28(5):361–4.Google Scholar
  10. 10.
    Lu X, Qin W, Li J, Tan N, Pan D, Zhang H, et al. The growth and metastasis of human hepatocellular carcinoma xenografts are inhibited by small interfering RNA targeting to the subunit ATP6L of proton pump. Cancer Res. 2005;65(15):6843–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Ma C, Liu Y, Zheng J, Fang W, You J, Wang J, et al. Identification of tumor metastasis related gene TMSG-1 by mRNA differential display. Sci China C Life Sci. 2002;45(5):553–60.CrossRefPubMedGoogle Scholar
  12. 12.
    Martinez-Zaguilan R, Seftor EA, Seftor RE, Chu YW, Gillies RJ, Hendrix MJ. Acidic pH enhances the invasive behavior of human melanoma cells. Clin Exp Metastasis. 1996;14(2):176–86.CrossRefPubMedGoogle Scholar
  13. 13.
    Mizutani Y, Kihara A, Igarashi Y. Mammalian Lass6 and its related family members regulate synthesis of specific ceramides. Biochem J. 2005;390(Pt1):263–71.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Montcourrier P, Mangeat PH, Valembois C, Salazar G, Sahuquet A, Duperray C, et al. Characterization of very acidic phagosomes in breast cancer cells and their association with invasion. J Cell Sci. 1994;107(Pt9):2381–91.PubMedGoogle Scholar
  15. 15.
    Ogretmen B, Hannun YA. Biologically active sphingolipids in cancer pathogenesis and treatment. Nat Rev Cancer. 2004;4:604–16.CrossRefPubMedGoogle Scholar
  16. 16.
    Pan H, Qin WX, Huo KK, Wan DF, Yu Y, Xu ZG, et al. Cloning, mapping, and characterization of a human homologue of the yeast longevity assurance gene LAG1. Genomics. 2001;77(1–2):58–64.CrossRefPubMedGoogle Scholar
  17. 17.
    Pei F, Ning JY, You JF, Yang JP, Wang YP, Han ZH, et al. Monoclonal antibodies against human tumor metastasis suppressor gene-1 (TMSG-1): preparation, characterization, and application. Hybrid Hybridomics. 2004;23:318–25.CrossRefGoogle Scholar
  18. 18.
    Rozhin J, Sameni M, Ziegler G, Sloane BF. Pericellular pH affects distribution and secretion of cathepsin B in malignant cells. Cancer Res. 1994;54(24):6517–25.PubMedGoogle Scholar
  19. 19.
    Schiffmann S, Sandner J, Birod K, Wobst I, Angioni C, Ruckhäberle E, et al. Ceramide synthases and ceramide levels are increased in breast cancer tissue. Carcinogenesis. 2009;30:745–52.CrossRefPubMedGoogle Scholar
  20. 20.
    Siegel R, Naishadham D, Jemal A. Cancer statistics, 2012. CA Cancer J Clin. 2012;62(1):10–29.CrossRefPubMedGoogle Scholar
  21. 21.
    Siegel R, Ward E, Brawley O, Jemal A. Cancer statistics, 2011: the impact of eliminating socioeconomic and racial disparities on premature cancer deaths. CA Cancer J Clin. 2011;61:212–36.CrossRefPubMedGoogle Scholar
  22. 22.
    Stiban J, Caputo L, Colombini M. Ceramide synthesis in the endoplasmic reticulum can permeabilize mitochondria to proapoptotic proteins. J Lipid Res. 2008;49(3):625–34.CrossRefPubMedGoogle Scholar
  23. 23.
    Strausberg RL, Feingold EA, Grouse LH, Derge JG, Klausner RD, Collins FS, et al. Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences. Proc Natl Acad Sci U S A. 2002;99(26):16899–903.CrossRefPubMedGoogle Scholar
  24. 24.
    Tang N, Jin J, Deng Y, Ke RH, Shen QJ, Fan SH, et al. LASS2 interacts with V-ATPase and inhibits cell growth of hepatocellular carcinoma. Acta Physiologica Sinica(Chinese). 2010;62(3):196–202.Google Scholar
  25. 25.
    Xu X, Liu B, Zou P, Zhang Y, You J, Pei F. Silencing of LASS2/TMSG1 enhances invasion and metastasis capacity of prostate cancer cell. J Cell Biochem. 2014;115:731–43.CrossRefPubMedGoogle Scholar
  26. 26.
    Xu XY, You JF, Pei F. Silencing of a novel tumor metastasis suppressor gene LASS2/TMSG1 promotes invasion of prostate cancer cell in vitro through increase of vacuolar ATPase activity. J Cell Biochem. 2012;113(7):2356–63.CrossRefPubMedGoogle Scholar
  27. 27.
    Yu W, Wang L, Wang Y, Xu X, Zou P, Gong M, et al. A novel tumor metastasis suppressor gene LASS2/TMSG1 interacts with vacuolar ATPase through its homeodomain. J Cell Biochem. 2012;114(3):570–83.CrossRefGoogle Scholar
  28. 28.
    Zigdon H, Kogot-Levin A, Park J-W, Goldschmidt R, Kelly S, Merrill Jr AH, et al. Ablation of ceramide synthase 2 causes chronic oxidative stress due to disruption of the mitochondrial respiratory chain*. J Biol Chem. 2013;288(7):4947–56.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Fang Mei
    • 1
  • Jiangfeng You
    • 1
  • Beiying Liu
    • 2
  • Mengxue Zhang
    • 1
  • Jiangying Liu
    • 1
  • Bo Zhang
    • 1
  • Fei Pei
    • 1
  1. 1.Department of Pathology, School of Basic Medical SciencesPeking University Health Science CenterBeijingChina
  2. 2.School of Mechanical EngineeringUniversity of Science & Technology BeijingBeijingChina

Personalised recommendations