Overexpression of FMNL2 is closely related to metastasis of colorectal cancer

Original Article

Abstract

Background and aims

Formin-like 2 (FMNL2) is a member of diaphanous-related formins which can control the actin-dependent processes such as cell motility and invasion. In this study, we investigated the expression of FMNL2 in colorectal cancer (CRC) and its correlation with CRC metastasis.

Patients–methods

Paraffin-embedded specimens of CRC (including 75 primary CRC tumors and 45 corresponding metastatic lymph nodes) and normal colorectal mucosa (30 samples) were immunostained with a FMNL2 antibody. Thirty-two paired snap-frozen tumor tissues and adjacent normal colorectal mucosa were subjected to real-time reverse-transcription polymerase chain reaction (RT-PCR). Six CRC cell lines (SW480, SW620, SW480/M5, LoVo, LS174T, and HT29) were assayed for FMNL2 expression by Western blotting and real-time RT-PCR. Their invasive abilities in vitro were determined by Boyden chamber assay.

Results

The immunohistochemical analysis showed FMNL2 expression was considerably higher in CRC tumors and corresponding metastatic lymph nodes than in normal colorectal mucosa (P < 0.05, respectively). Elevated FMNL2 expression was significantly correlated with lymphatic metastasis of CRC (P < 0.05). Real-time RT-PCR analysis confirmed the results obtained by immunohistochemistry. At mRNA and protein levels, FMNL2 expression was substantially upregulated in cell lines derived from CRC metastases (SW620, SW480/M5, and LoVo) compared to ones derived from primary CRC (HT29, LS174T, and SW480; P < 0.05). In vitro cell invasive assay demonstrated that the former three cell lines had higher invasive ability than the latter cell lines.

Conclusions

FMNL2 may play an important role in the metastasis of CRC and may be a useful marker for metastasis of CRC.

Keywords

Formin-like 2 (FMNL2) Diaphanous-related formins (DRFs) Colorectal cancer (CRC) Metastasis Invasion 

Abbreviations

FMNL2

formin-like 2

CRC

colorectal cancer

IHC

immunohistochemistry

DRFs

diaphanous-related formins

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China Nos. 30400206, 30370649, 30670967, and 30770977.

References

  1. 1.
    Jemal A, Murray T (2005) Ward ECancer statistics. CA Cancer J Clin 55:10–30PubMedCrossRefGoogle Scholar
  2. 2.
    Millikan KW, Staren ED, Doolas A (1997) Invasive therapy of metastatic colorectal cancer to the liver. Surg Clin North Am 77:27–48PubMedCrossRefGoogle Scholar
  3. 3.
    Zhang YF, Liu L, Ding YQ (2007) Isolation and characterization of human colorectal cancer cell subline with unique metastatic potential in the liver. Nan Fang Yi Ke Da Xue Xue Bao 27:126–130PubMedGoogle Scholar
  4. 4.
    Goode BL, Eck MJ (2007) Mechanism and function of formins in the control of actin assembly. Annu Rev Biochem 76:593–627PubMedCrossRefGoogle Scholar
  5. 5.
    Higgs HN (2005) Formin proteins: a domain-based approach. Trends Biochem Sci 30:342–353PubMedCrossRefGoogle Scholar
  6. 6.
    Faix J, Grosse R (2006) Staying in shape with formins. Dev Cell 10:693–706PubMedCrossRefGoogle Scholar
  7. 7.
    Eisenmann KM, Harris ES, Kitchen SM, Holman HA, Higgs HN, Alberts AS (2007) Dia-interacting protein modulates formin-mediated actin assembly at the cell cortex. Curr Biol 17:579–591PubMedCrossRefGoogle Scholar
  8. 8.
    Sahai E (2005) Mechanisms of cancer cell invasion. Curr Opin Genet Dev 15:87–96PubMedCrossRefGoogle Scholar
  9. 9.
    Wodarz A, Näthke I (2007) Cell polarity in development and cancer. Nat Cell Biol 9:1016–1024PubMedCrossRefGoogle Scholar
  10. 10.
    Vasiliev JM (2004) Cytoskeletal mechanisms responsible for invasive migration of neoplastic cells. Int J Dev Biol 48:425–439PubMedCrossRefGoogle Scholar
  11. 11.
    Watanabe N, Madaule P, Reid T, Ishizaki T, Watanabe G, Kakizuka A, Saito Y, Nakao K, Jockusch BM, Narumiya S (1997) p140mDia, mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin. EMBO J 16:3044–3056PubMedCrossRefGoogle Scholar
  12. 12.
    Kitzing TM, Sahadevan AS, Brandt DT, Knieling H, Hannemann S, Fackler OT, Grosshans J, Grosse R (2007) Positive feedback between Dia1, LARG, and RhoA regulates cell morphology and invasion. Genes Dev 21:1478–1483PubMedCrossRefGoogle Scholar
  13. 13.
    Soumaoro LT, Uetake H, Higuchi T, Takagi Y, Enomoto M, Sugihara K (2004) Cyclooxygenase-2 expression: a significant prognostic indicator for patients with colorectal cancer. Clin Cancer Res 10:8465–8471PubMedCrossRefGoogle Scholar
  14. 14.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2-Delta Delta CT Method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  15. 15.
    Itoh F, Yamamoto H, Hinoda Y, Imai K (1996) Enhanced secretion and activation of matrilysin during malignant conversion of human colorectal epithelium and its relationship with invasive potential of colon cancer cells. Cancer 77:1717–1721PubMedGoogle Scholar
  16. 16.
    Mlodzik M (2002) Planar cell polarization: do the same mechanisms regulate Drosophila tissue polarity and vertebrate gastrulation? Trends Genet 18:564–571PubMedCrossRefGoogle Scholar
  17. 17.
    Pellegrin S, Mellor H (2005) The Rho family GTPase Rif induces filopodia through mDia2. Curr Biol 15:129–133PubMedCrossRefGoogle Scholar
  18. 18.
    Katoh M, Katoh M (2003) Identification and characterization of human FMNL1, FMNL2 and FMNL3 genes in silico. Int J Oncol 22:1161–1168PubMedGoogle Scholar
  19. 19.
    Rivero F, Muramoto T, Meyer AK, Urushihara H, Uyeda TQ, Kitayama C (2005) A comparative sequence analysis reveals a common GBD/FH3-FH1-FH2-DAD architecture in formins from Dictyostelium, fungi and metazoa. BMC Genomics 6:28PubMedCrossRefGoogle Scholar
  20. 20.
    Alberts AS (2001) Identification of a carboxyl-terminal diaphanous-related formin homology protein autoregulatory domain. J Biol Chem 276:2824–2830PubMedCrossRefGoogle Scholar
  21. 21.
    Rose R, Weyand M, Lammers M, Ishizaki T, Ahmadian MR, Wittinghofer A (2005) Structural and mechanistic insights into the interaction between Rho and mammalian Dia. Nature 435:513–518PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.Department of PathologySouthern Medical UniversityGuangzhouChina

Personalised recommendations