Functional analysis of Zyxin in cell migration and invasive potential of oral squamous cell carcinoma cells Corrigendum in /10.3892/ijo.2016.3702

Yamamura,Michiyo; Noguchi,Kazuma; Nakano,Yoshiro; Segawa,Emi; Zushi,Yusuke; Takaoka,Kazuki; Kishimoto,Hiromitsu; Hashimoto-Tamaoki,Tomoko; Urade,Masahiro

doi:10.3892/ijo.2013.1761

Functional analysis of Zyxin in cell migration and invasive potential of oral squamous cell carcinoma cells

Corrigendum in: /10.3892/ijo.2016.3702

Authors:
- Michiyo Yamamura
- Kazuma Noguchi
- Yoshiro Nakano
- Emi Segawa
- Yusuke Zushi
- Kazuki Takaoka
- Hiromitsu Kishimoto
- Tomoko Hashimoto-Tamaoki
- Masahiro Urade
View Affiliations

Affiliations: Department of Oral and Maxillofacial Surgery, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan, Department of Genetics, Hyogo College of Medicine, Nishinomiya, Hyogo 663-8501, Japan
Published online on: January 4, 2013 https://doi.org/10.3892/ijo.2013.1761
Pages: 873-880

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Zyxin is an evolutionarily conserved protein that has been implicated in the regulation of actin assembly and is mainly located at focal adhesions. However, the biological roles of Zyxin in cancer cells are incompletely understood. We analyzed the functions of Zyxin in cell migration and the invasive potential of OSCC. Zyxin expression was examined using eight OSCC cell lines with two different cell morphologies (6 epithelial type and 2 fibroblastic type). To knockdown Zyxin expression, OSCC cells were transfected with Zyxin siRNA and control siRNA. The cell lines were studied by western blot analysis, immunocytochemical analysis and cell migration and invasion assay. Epithelial type OSCC cells showed a high level of E-cadherin expression and a low level of Zyxin expression. N-cadherin as well as Zyxin were strongly expressed in fibroblastic type OSCC cells. Expression levels of LPP and TRIP6, members of the human Zyxin family, did not differ between epithelial type and fibroblastic type. Knockdown of Zyxin expression by siRNA in fibroblastic type OSCC cells was associated with cell morphological changes from spindle (fibroblastic) to polygonal (epithelial) shape and significantly inhibited cell growth as well as cell migration and invasion. Expression levels of Rac1 and Cdc42 were weaker in Zyxin siRNA-treated fibroblastic type OSCC cells than in control siRNA-treated cells, but the expression of RhoA did not differ significantly. Treatment of fibroblastic type OSCC cells with Rac1 inhibitor decreased the expression of Zyxin mRNA and protein. Zyxin is suggested to promote growth, migration and invasiveness of fibroblastic type OSCC cells by upregulating Rac1 and Cdc42.

View Figures

View References

1	Pereira MC, Oliveira DT, Landman G and Kowalski LP: Histologic subtypes of oral squamous cell carcinoma: prognostic relevance. J Can Dent Assoc. 73:339–344. 2007.PubMed/NCBI
2	Sudbo J, Bryne M, Mao L, Lotan R, et al: Molecular based treatment of oral cancer. Oral Oncol. 39:749–758. 2003. View Article : Google Scholar
3	Leemans CR, Tiwari R, Nauta JJ, van der Waal I and Snow GB: Regional lymph node involvement and significance in the development of distant metastases in head and neck carcinoma. Cancer. 71:452–456. 1993. View Article : Google Scholar : PubMed/NCBI
4	Leemans CR, Tiwari R, Nauta JJ, van der Waal I and Snow GB: Recurrence at the primary site in head and neck cancer and the significance of neck lymph node metastases as a prognostic factor. Cancer. 73:187–190. 1994. View Article : Google Scholar : PubMed/NCBI
5	Brattain MG, Willson J, Koterba A, Patil S and Venkateswarlu S: Colorectal cancer. Human Cell Culture. Cancer Cell Lines, Part 2. Masters JR and Palsson B: 2. Kluwer Academic Publishers; Norwell, MA: pp. 293–304. 2002
6	Kozlowski J and Sensibar JA: Prostate cancer. Human Cell Culture. Cancer Cell Lines, Part 2. Masters JR and Palsson B: 2. Kluwer Academic Publishers; Norwell, MA: pp. 305–332. 2002
7	Sutherland R, Watts CK, Lee CS and Musgrove EA: Breast cancer. Human Cell Culture. Cancer Cell Lines, Part 2. Masters JR and Palsson B: 2. Kluwer Academic Publishers; Norwell, MA: pp. 79–106. 2002
8	Yokoyama K, Kamata N, Hayashi E, Hoteiya T, Ueda N, Fujimoto R and Nagayama M: Reverse correlation of E-cadherin and snail expression in oral squamous cell carcinoma cells in vitro. Oral Oncol. 37:65–71. 2001. View Article : Google Scholar : PubMed/NCBI
9	Frixen UH, Behrens J, Sachs M, et al: E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells. J Cell Biol. 113:173–185. 1991. View Article : Google Scholar : PubMed/NCBI
10	Petit MM, Fradelizi J, Golsteyn RM, et al: LPP, an actin cytoskeleton protein related to zyxin, harbors a nuclear export signal and transcriptional activation capacity. Mol Biol Cell. 11:117–129. 2000. View Article : Google Scholar : PubMed/NCBI
11	Yi J and Beckerle MC: The human TRIP6 gene encodes a LIM domain protein and maps to chromosome 7q22, a region associated with tumorigenesis. Genomics. 49:314–316. 1998. View Article : Google Scholar : PubMed/NCBI
12	Grunewald TG, Kammerer U, Schulze E, Schindler D, Hong A, Zimmer M and Butt E: Silencing of LASP-1 influences zyxin localization, inhibits proliferation and reduces migration in breast cancer cells. Exp Cell Res. 312:974–982. 2006. View Article : Google Scholar : PubMed/NCBI
13	Grunewald TG, Kammerer U, Winkler C, Schindler D, Sickmann A, Honig A and Butt E: Overexpression of LASP-1 mediates migration and proliferation of human ovarian cancer cells and influences zyxin localisation. Br J Cancer. 96:296–305. 2007. View Article : Google Scholar : PubMed/NCBI
14	Hervy M, Hoffman L and Beckerle MC: From the membrane to the nucleus and back again: bifunctional focal adhesion proteins. Curr Opin Cell Biol. 18:524–532. 2006. View Article : Google Scholar : PubMed/NCBI
15	Nix DA and Beckerle MC: Nuclear-cytoplasmic shuttling of the focal contact protein, zyxin: a potential mechanism for communication between sites of cell adhesion and the nucleus. J Cell Biol. 138:1139–1147. 1997. View Article : Google Scholar
16	Nix DA, Fradelizi J, Bockholt S, Menichi B, Louvard D, Friedich E and Beckerle MC: Targeting of zyxin to sites of actin membrane interaction and to the nucleus. J Biol Chem. 276:34759–34767. 2001. View Article : Google Scholar : PubMed/NCBI
17	van der Gaag EJ, Leccica MT, Dekker SK, Jalbert NL, Amodeo DM and Byers HR: Role of Zyxin in differential cell spreading and proliferation of melanoma cells and melanocytes. J Invest Dermatol. 118:246–254. 2002.PubMed/NCBI
18	Sperry RB, Bishop SN, Bramwell JJ, et al: Zyxin controls migration in epithelial-mesenchymal transition by mediating actin-membrane linkages at cell-cell junctions. J Cell Physiol. 222:612–624. 2010.PubMed/NCBI
19	Urade M, Sugi M and Miyazaki T: Establishment of three bleomycin-resistant human carcinoma cell lines and their cross resistance to other antitumor agents. Cancer. 61:1501–1507. 1998. View Article : Google Scholar
20	Kamata N, Chida K, Rikimaru K, Horikoshi M, Enomoto S and Kuroki T: Growth inhibitory effects of epidermal growth factor and overexpression of its receptors on human squamous cell carcinomas in culture. Cancer Res. 46:1648–1653. 1986.PubMed/NCBI
21	Kawahara E, Okada Y, Nakanishi I, Iwata K, Kojima S, Kumagai S and Yamamoto E: The expression of invasive behavior of differentiated squamous carcinoma cell line evaluated by an in vitro invasion model. Jpn J Cancer Res. 84:409–418. 1993. View Article : Google Scholar : PubMed/NCBI
22	Kojima S: Experimenral study of invasive activity of oral squamous cell carcinoma. J Juzen Med Soc. 101:266–281. 1992.
23	Hoteiya T, Hayashi E, Satomura K, Kamata N and Nagayama N: Expression of E-cadherin in oral cancer cell lines and its relationship to invasiveness in SCID mice in vivo. J Oral Pathol Med. 28:107–111. 1999. View Article : Google Scholar : PubMed/NCBI
24	Kawashiri S, Tanaka A, Noguchi N, et al: Invasion and metastasis models of oral squamous carcinoma cell lines. Jpn J Tissue Cult Dent Res. 12:1–13. 2004.(in Japanese).
25	Moriyama M: Development of diffuse invasive (grade4D) human oral squamous cell carcinoma model in severe combined immunodeficiency mice: microangioarchitectual analysis and immunohistochemical study. Oral Oncol. 35:395–400. 1999. View Article : Google Scholar : PubMed/NCBI
26	Hashitani S, Urade M, Nishimura N, Maeda N, Takaoka K, Noguchi K and Sakurai K: Apoptosis induction enhancement of anticancer drugs by celecoxib, a selective cyclooxygenase-2 inhibitor, in human head neck carcinoma cell lines. Int J Oncol. 23:665–672. 2003.
27	Hiromoto T, Noguchi K, Yamamura M, et al: Up-regulation of neurophil gelatinase-associated lipocalin in oral squaamous cell carcinoma: relation to cell differentiation. Oncol Rep. 26:1415–1421. 2011.PubMed/NCBI
28	Higashikawa K, Yoneda S, Tobiume K, et al: ΔNp63α-dependent expression of Id-3 distinctively suppresses the invasiveness of human squamous cell carcinoma. Int J Cancer. 124:2837–2844. 2009.
29	Bendris N, Arsic N, Lemmers B and Bianchard JM: Cyclin A2, Rho GTPases and EMT. Small GTPases. Oct 1–2012.(Epub ahead of print).
30	Diniz-Freitas M, García-Caballero T, Antúnez-López J, Gándara-Rey JM and García-García A: Reduced E-cadherin expression is an indicator of unfavourable prognosis in oral squamous cell carcinoma. Oral Oncol. 42:190–200. 2006. View Article : Google Scholar : PubMed/NCBI
31	Tanaka N, Odajima T, Ogi K, Ikeda T and Satoh M: Expression of E-cadherin, alpha-catenin, and beta-catenin in the process of lymph node metastasis in oral squamous cell carcinoma. Br J Cancer. 89:557–563. 2003. View Article : Google Scholar : PubMed/NCBI
32	Krisanaprakornkit S and Iamaroon A: Epithelial-mesenchymal transition in oral squamous cell carcinoma. ISRN Oncol. 681469. 2012. View Article : Google Scholar
33	Fradelizi j, Noireaux V, Plastino J, et al: ActA and human zyxin harbor Arp2/3-independent actin-polymerization activity. Nat Cell Biol. 3:699–707. 2001. View Article : Google Scholar : PubMed/NCBI
34	Beckerle MC: Spatial control of action filament assembly: lessons from Listeria. Cell. 95:741–748. 1998. View Article : Google Scholar : PubMed/NCBI
35	Kadrmas JL and Beckerle MC: The LIM domain: from the cytoskeleton to the nucleus. Nat Rev Mol Cell Biol. 5:920–931. 2004. View Article : Google Scholar : PubMed/NCBI
36	Reiling JH and Sabatini DM: Stress and mTORture signaling. Oncogene. 25:6373–6383. 2006. View Article : Google Scholar : PubMed/NCBI
37	Shinto O, Yashiro M, Kawajiri H, Shimizu K, Shimizu T, Miwa A and Hirakawa K: Inhibitory effect of a TGFbeta receptor type-I inhibitor, Ki26894, on invasiveness of scirrhous gastric cancer cells. Br J Cancer. 102:844–851. 2010. View Article : Google Scholar : PubMed/NCBI
38	Amsellem V, Kryszke MH, Hervy M, et al: The actin cytoskeleton-associated protein zyxin acts as a tumor suppressor in Ewing tumor cells. Exp Cell Res. 304:443–456. 2005. View Article : Google Scholar : PubMed/NCBI
39	Wendt MK, Allington TM and Schiemann WP: Mechanisms of the epithelial-mesenchymal transition by TGF-beta. Future Oncol. 5:1145–1168. 2009. View Article : Google Scholar : PubMed/NCBI
40	Pratt S J, Epple H, Ward M, Feng Y, Braga VM and Longmore GD: The LIM protein Ajuba influences p130Cas localization and Rac1 activity during cell migration. J Cell Biol. 168:813–824. 2005.PubMed/NCBI