Effect of hypoglycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma

Abstract

Abstract: PURPOSE: To investigate the effect of N-glycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma CAL 27 cells, and to study the effect of hypoglycosylation of E-cadherins on the stability of adherens junctions (AJs) mediated by E-cadherins. METHODS: Human tongue squamous cell carcinoma CAL 27 cells were respectively transfected with the plasmids encoding either hypoglycosylated or wild-type E-cadherins with FLAG as tags. Western blot of FLAG was used to detect the expression of exogenous E-cadherins in the transfected cells after 48 hours. The cell proliferation counting, monolayer cell scratch wound healing and in vitro cell invasiveness assays were performed to evaluate the proliferation and invasiveness of cells. Immunoprecipitation experiments were used to assay the quantity of α-catenins, β-catenins, γ-catenins and vinculins linked with the cytosolic tails of E-cadherins. All statistical analysis were performed using SPSS 17.0 software package. RESULTS: The expression of exogenous E-cadherins was detected in transfected CAL 27 cells by Western blot. Cell proliferation counting assay revealed that hypoglycosylated E-cadherins significantly inhibited proliferation of CAL 27 cells, compared with wild-type E-cadherins. The monolayer cell scratch wound healing and in vitro cell invasiveness assays indicated that hypoglycosylated E-cadherins also significantly restrained both invasiveness of CAL 27 cells, compared with wild-type E-cadherins. The immunoprecipitation experiments demonstrated that hypoglycosylated E-cadherins exhibited an increased association with α-catenins, β-catenins, and γ-catenins and vinculins. CONCLUSIONS: Compared with wild-type E-cadherins, hypoglycosylated E-cadherins can significantly suppress proliferation and invasiveness of tongue squamous cell carcinoma CAL 27 cells, and the mechanism was that hypoglycosylated E-cadherins can induce more stable AJs than wild-type E-cadherin.

Key words: Glycosylation, E-cadherin, Tongue squamous cell carcinoma neoplasms, Cell proliferation, Neoplasm invasiveness, Adherens junctions

CLC Number:

R739.8

ZHANG Ping-ping1, XU Xiu-ying1, GAO Zhen-nan1. Effect of hypoglycosylated E-cadherins on proliferation and invasiveness of tongue squamous cell carcinoma[J]. Shanghai Journal of Stomatology, 2014, 23(1): 1-6.

References

[1] Christofori G. New signals from the invasive front[J]. Nature, 2006, 441(7092): 444-450.
[2] Baranwal S, Alahari SK. Molecular mechanisms controlling E-cadherins expression in breast cancer [J]. Biochem Biophys Res Commun, 2009, 384(1): 6-11.
[3] Hart GW, Copeland RJ. Glycomics hits the big time [J]. Cell, 2010, 143(5): 672-676.
[4] Nita-Lazar M, Noonan V, Rebustini I, et al. Overexpression of DPAGT1 leads to aberrant N-glycosylation of E-cadherins and cellular discohesion in oral cancer [J]. Cancer Res, 2009, 69(14): 5673-5680.
[5] Liwosz A, Lei T, Kukuruzinska MA. N-glycosylation affects the molecular organization and stability of E-cadherins junctions [J]. J Biol Chem, 2006, 281(32): 23138-23149.
[6] 郑家伟, 李金忠, 钟来平, 等. 口腔鳞状细胞癌临床流行病学研究现状 [J]. 中国口腔颌面外科杂志, 2007, 5(2): 83-90.
[7] Hugo HJ, Kokkinos MI, Blick T, et al. Defining the E-cadherins repressor interactome in epithelial-mesenchymal transition: the PMC42 model as a case study[J]. Cells Tissues Organs, 2011, 193(1-2): 23-40.
[8] Thiery JP. Epithelial-mesenchymal transitions in tumour progression [J]. Nat Rev Cancer, 2002, 2(6): 442-454.
[9] Espejo R, Rengifo-Cam W, Schaller MD, et al. PTP-PEST controls motility, adherens junction assembly, and Rho GTPase activity in colon cancer cells[J]. Am J Physiol Cell Physiol, 2010, 299(2): C454-463.
[10] Mortazavi F, Dubinett S, Rettig M. c-Crk proto-oncogene contributes to transcriptional repression of p120-catenins in non-small cell lung cancer cells [J]. Clin Exp Metastasis, 2011, 28(4): 391-404.
[11] Dennis JW, Granovsky M, Warren CE. Glycoprotein glycosylation and cancer progression [J]. Biochim Biophys Acta, 1999, 1473(1): 21-34.
[12] Jamora C, Fuchs E. Intercellular adhesion, signalling and the cytoskeleton [J]. Nat Cell Biol, 2002, 4(4): E101-108.
[13] Pinho SS, Seruca R, Gärtner F, et al. Modulation of E-cadherins function and dysfunction by N-glycosylation [J]. Cell Mol Life Sci, 2011, 68(6): 1011-1020.
[14] Zhao YY, Takahashi M, Gu JG, et al. Functional roles of N-glycans in cell signaling and cell adhesion in cancer [J]. Cancer Sci, 2008, 99(7): 1304-1310.
[15] Freeze HH. Genetic defects in the human glycome [J]. Nat Rev Genet, 2006, 7(7): 537-551.
[16] Cavallaro U, Christofori G. Cell adhesion and signalling by cadherins and Ig-CAMs in cancer[J]. Nat Rev Cancer,2004, 4(2): 118-132.
[17] Lewis-Tuffin LJ, Rodriguez F, Giannini C, et al. Misregulated E-cadherins expression associated with an aggressive brain tumor phenotype [J]. PLoS One, 2010, 5(10): e13665.
[18] Lau MT, Klausen C, Leung PC. E-cadherins inhibits tumor cell growth by suppressing PI3K/Akt signaling via β-catenins-Egr1-mediated PTEN expression [J]. Oncogene, 2011, 30(24): 2753-2766.
[19] de Freitas Junior JC, Silva Bdu R, de Souza WF, et al. Inhibition of N-linked glycosylation by tunicamycin induces E-cadherin-mediated cell-cell adhesion and inhibits cell proliferation in undifferentiated human colon cancer cells [J]. Cancer Chemother Pharmacol, 2011, 68(1): 227-238.