Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Cancer Cell International
Published in: Cancer Cell International 1/2008

Open Access 01-12-2008 | Primary research

Reduced paxillin expression contributes to the antimetastatic effect of 4-hydroxycoumarin on B16-F10 melanoma cells

Authors: Marco A Velasco-Velázquez, Nohemí Salinas-Jazmín, Nicandro Mendoza-Patiño, Juan J Mandoki

Published in: Cancer Cell International | Issue 1/2008

Login to get access

Abstract

Background

4-Hydroxycoumarin (4-HC) is a coumarin that lacks anticoagulant activity. 4-HC affects the cytoskeletal stability and decreases cell adhesion and motility of the melanoma cell line B16-F10. Together with integrins and other cytoskeletal proteins, paxillin participates in the regulation of cell adhesion and motility, acting as an adapter protein at focal adhesions. The present study determined the participation of paxillin in the reported effects of 4-HC and analyzed the role of paxillin in the formation of melanoma metastases.

Results

4-HC decreased protein and mRNA levels of α- and β-paxillin isoforms in B16-F10 cells. Paxillin downregulation correlated with an inadequate translocation of paxillin to focal adhesions and a reduced phosphotyr118-paxillin pool. Consequently, 4-HC altered paxillin-mediated signaling, decreasing the phosphorylation of FAK and the level of GTP-bound Rac-1. These results partially explain the mechanism of the previously reported effects of 4-HC. Additionally, we studied the effect of 4-HC on metastatic potential of B16-F10 cells through experimental metastasis assays. In vitro treatment of cells with 4-HC inhibited their capability to originate pulmonary metastases. 4-HC did not affect cell proliferation or survival, demonstrating that its antimetastatic effect is unrelated to changes on cell viability. We also studied the importance of paxillin in metastasis by transfecting melanoma cells with paxillin-siRNA. Transfection produced a modest reduction on metastatic potential, indicating that: i) paxillin plays a role as inducer of melanoma metastasis; and ii) paxillin downregulation is not sufficient to explain the antimetastatic effect of 4-HC. Therefore, we evaluated other changes in gene expression by differential display RT-PCR analysis. Treatment with 4-HC produced a downregulation of Adhesion Regulating Molecule-1 (ARM-1), which correlated with a decreased adhesion of melanoma cells to lung slides.

Conclusion

This study shows that reduced paxillin expression is associated with the impaired cell adhesion and motility seen in 4-HC-treated cells and partially contributes to the antimetastatic effect of 4-HC. In contrast, the role of ARM-1 reduced expression in the effects of 4-HC is still to be clarified. The antimetastatic effect of 4-HC suggests that this compound, or others with similar mode of action, might be useful for the development of adjuvant therapies for melanoma.
Appendix
Available only for authorised users
Literature
1.
Bogenrieder T, Herlyn M: Axis of evil: molecular mechanisms of cancer metastasis. Oncogene. 2003, 22: 6524-6536. 10.1038/sj.onc.1206757.CrossRefPubMed
2.
Hood JD, Cheresh DA: Role of integrins in cell invasion and migration. Nat Rev Cancer. 2002, 2: 91-100. 10.1038/nrc727.CrossRefPubMed
3.
Geiger B, Bershadsky A, Pankov R, Yamada K: Transmembrane extracellular matrix-cytoskeleton crosstalk. Nat Rev Mol Cell Biol. 2001, 2: 793-805. 10.1038/35099066.CrossRefPubMed
4.
Ridley AJ, Schwartz MA, Burridge K, Firtel RA, Ginsberg MH, Borisy G, Parsons JT, Horwitz AR: Cell migration: integrating signals from front to back. Science. 2003, 302: 1704-1709. 10.1126/science.1092053.CrossRefPubMed
5.
Hehlgans S, Haase M, Cordes N: Signaling via integrins: implications for cell survival and anticancer strategies. Biochim Biophys Acta. 2007, 1775: 163-180.PubMed
6.
Velasco-Velázquez MA, Molina-Guarneros JA, Mendoza-Patiño N, López-González JS, Mandoki JJ: Integrins and integrin-associated molecules: targets for the development of antimetastatic therapies. Rev Invest Clin. 1999, 51: 183-93.PubMed
7.
Brown MC, Turner CE: Paxillin: adapting to change. Physiol Rev. 2004, 84: 1315-1339. 10.1152/physrev.00002.2004.CrossRefPubMed
8.
Wang Y, Gilmore TD: Zyxin and paxillin proteins: focal adhesion plaque LIM domain proteins go nuclear. Biochim Biophys Acta. 2003, 1593: 115-120. 10.1016/S0167-4889(02)00349-X.CrossRefPubMed
9.
Burridge K, Turner CE, Romer LH: Tyrosine phosphorylation of paxillin and pp125FAK accompanies cell adhesion to extracellular matrix: A role in cytoskeletal assembly. J Cell Biol. 1992, 119: 893-903. 10.1083/jcb.119.4.893.CrossRefPubMed
10.
Sanders MA, Basson MD: Collagen IV-dependent ERK activation in human Caco-2 intestinal epithelial cells requires focal adhesion kinase. J Biol Chem. 2000, 275: 38040-38047. 10.1074/jbc.M003871200.CrossRefPubMed
11.
Jamieson JS, Tumbarello DA, Halle M, Brown MC, Tremblay ML, Turner CE: Paxillin is essential for PTP-PEST-dependent regulation of cell spreading and motility: a role for paxillin kinase linker. J Cell Sci. 2005, 118: 5835-5847. 10.1242/jcs.02693.CrossRefPubMed
12.
Petit V, Boyer B, Lentz D, Turner CE, Thiery JP, Valles AM: Phosphorylation of tyrosine residues 31 and 118 on paxillin regulates cell migration through an association with CRK in NBT-II cells. J Cell Biol. 2000, 148: 957-970. 10.1083/jcb.148.5.957.PubMedCentralCrossRefPubMed
13.
Mazaki Y, Hashimoto S, Sabe H: Monocyte cells and cancer cells express novel paxillin isoforms with different binding properties to focal adhesion proteins. J Biol Chem. 1997, 272: 7437-7444. 10.1074/jbc.272.11.7437.CrossRefPubMed
14.
Mazaki Y, Hiroshi U, Okio H, Shigeru H: Paxillin Isoform in Mouse. Lack of the gamma isoform and developmentally specific beta isoform expression. J Biol Chem. 1998, 273: 22435-22441. 10.1074/jbc.273.35.22435.CrossRefPubMed
15.
Richardson AR, Malik K, Hildebrand JD, Parsons JT: Inhibition of cell spreading by expression of the C-terminal domain of focal adhesion kinase (FAK) is rescued by coexpression of Src or catalytically inactive FAK: a role for paxillin tyrosine phosphorylation. Mol Cell Biol. 1997, 17: 6906-6914.PubMedCentralCrossRefPubMed
16.
Turner CE: Paxillin and focal adhesion signaling. Nat Cell Biol. 2000, 2: E231-E236. 10.1038/35046659.CrossRefPubMed
17.
Nakamura K, Yano H, Uchida H, Hashimoto S, Schaefer E, Sabe H: Tyrosine phosphorylation of paxillin alpha is involved in temporospatial regulation of paxillin-containing focal adhesion formation and F-actin organization in motile cells. J Biol Chem. 2000, 275: 27155-27164.PubMed
18.
Tsubouchi A, Sakakura J, Yagi R, Mazaki Y, Schaefer E, Yano H, Sabe H: Localized suppression of RhoA activity by Tyr31/118-phosphorylated paxillin in cell adhesion and migration. J Cell Biol. 2002, 159: 673-683. 10.1083/jcb.200202117.PubMedCentralCrossRefPubMed
19.
Iwasaki T, Nakata A, Mukai M, Shinkai K, Yano H, Sabe H, Schaefer E, Tatsuta M, Tsujimura T, Terada N, Kakishita E, Akedo H: Involvement of phosphorylation of Tyr-31 and Tyr-118 of paxillin in MM1 cancer cell migration. Int J Cancer. 2002, 97: 330-335. 10.1002/ijc.1609.CrossRefPubMed
20.
Conway WC, van Zyp Van der Voort J, Thamilselvan V, Walsh MF, Crowe DL, Basson MD: Paxillin modulates squamous cancer cell adhesion and is important in pressure-augmented adhesion. J Cell Biochem. 2006, 98: 1507-1516. 10.1002/jcb.20819.CrossRefPubMed
21.
Crowe DL, Ohannessian A: Recruitment of focal adhesion kinase and paxillin to beta1 integrin promotes cancer cell migration via mitogen activated protein kinase activation. BMC Cancer. 2004, 4: 18-10.1186/1471-2407-4-18.PubMedCentralCrossRefPubMed
22.
Azuma K, Tanaka M, Uekita T, Inoue S, Yokota J, Ouchi Y, Sakai R: Tyrosine phosphorylation of paxillin affects the metastatic potential of human osteosarcoma. Oncogene. 2005, 24: 4754-4764. 10.1038/sj.onc.1208654.CrossRefPubMed
23.
Jenq W, Cooper D, Ramirez G: Integrin expression on cell adhesion function and up-regulation of P125FAK and paxillin in metastatic renal carcinoma cells. Connect Tissue Res. 1996, 34: 161-174. 10.3109/03008209609000696.CrossRefPubMed
24.
Hamamura K, Furukawa K, Hayashi T, Hattori T, Nakano J, Nakashima H, Okuda T, Mizutani H, Hattori H, Ueda M, Urano T, Lloyd KO, Furukawa K: Ganglioside GD3 promotes cell growth and invasion through p130Cas and paxillin in malignant melanoma cells. Proc Natl Acad Sci USA. 2005, 102: 11041-11046. 10.1073/pnas.0503658102.PubMedCentralCrossRefPubMed
25.
Mueller RL, Scheidt S: History of drugs for thrombotic disease. Discovery, development, and directions for the future. Circulation. 1994, 89: 432-449.CrossRefPubMed
26.
Velasco-Velazquez MA, Agramonte-Hevia J, Barrera D, Jimenez-Orozco A, Garcia-Mondragon MJ, Mendoza-Patino N, Landa A, Mandoki J: 4-Hydroxycoumarin disorganizes the actin cytoskeleton in B16-F10 melanoma cells but not in B82 fibroblasts, decreasing their adhesion to extracellular matrix proteins and motility. Cancer Lett. 2003, 198: 179-186. 10.1016/S0304-3835(03)00333-1.CrossRefPubMed
27.
28.
Chacón-Salinas R, Serafín-López J, Ramos-Payán R, Méndez-Aragón P, Hernández-Pando R, Van Soolingen D, Flores-Romo L, Estrada-Parra S, Estrada-García I: Differential pattern of cytokine expression by macrophages infected in vitro with different Mycobacterium tuberculosis genotypes. Clin Exp Immunol. 2005, 140: 443-449. 10.1111/j.1365-2249.2005.02797.x.PubMedCentralCrossRefPubMed
29.
Ramos-Payán R, Aguilar-Medina M, Estrada-Parra S, González-Y-Merchand JA, Favila-Castillo L, Monroy-Ostria A, Estrada-García IC: Quantification of cytokine gene expression using an economical real-time polymerase chain reaction method based on SYBR Green I. Scand J Immunol. 2003, 57: 439-445. 10.1046/j.1365-3083.2003.01250.x.CrossRefPubMed
30.
Babich H, Shopsis C, Borenfreund E: In vitro cytotoxicity testing of aquatic pollutants (cadmium, copper, zinc, nickel) using established fish cell lines. Ecotoxicol Environ Saf. 1986, 11: 91-99. 10.1016/0147-6513(86)90030-8.CrossRefPubMed
31.
Franken NA, Rodermond HM, Stap J, Haveman J, van Bree C: Clonogenic assay of cells in vitro. Nat Protoc. 2006, 1: 2315-2319. 10.1038/nprot.2006.339.CrossRefPubMed
32.
Liang P, Pardee AB: Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science. 1992, 257: 967-971. 10.1126/science.1354393.CrossRefPubMed
33.
Vink J, Thomas L, Etoh T, Bruijn JA, Mihm MC, Gattoni-Celli S, Byers HR: Role of beta-1 integrins in organ specific adhesion of melanoma cells in vitro. Lab Invest. 1993, 68: 192-203.PubMed
34.
Schroeder MJ, Webb DJ, Shabanowitz J, Horwitz AF, Hunt DF: Methods for the detection of paxillin post-translational modifications and interacting proteins by mass spectrometry. J Proteome Res. 2005, 4: 1832-1841. 10.1021/pr0502020.CrossRefPubMed
35.
Abdel-Ghany M, Cheng HC, Elble RC, Pauli BU: Focal adhesion kinase activated by beta(4) integrin ligation to mCLCA1 mediates early metastatic growth. J Biol Chem. 2002, 277: 34391-34400. 10.1074/jbc.M205307200.CrossRefPubMed
36.
Gabarra-Niecko V, Schaller MD, Dunty JM: FAK regulates biological processes important for the pathogenesis of cancer. Cancer Metastasis Rev. 2003, 22: 359-374. 10.1023/A:1023725029589.CrossRefPubMed
37.
Simins AB, Weighardt H, Weidner KM, Weidle UH, Holzmann B: Functional cloning of ARM-1, an adhesion-regulating molecule upregulated in metastatic tumor cells. Clin Exp Metastasis. 1999, 17: 641-648. 10.1023/A:1006790912877.CrossRefPubMed
38.
Cohen LA, Guan JL: Mechanisms of focal adhesion kinase regulation. Curr Cancer Drug Targets. 2005, 5: 629-643. 10.2174/156800905774932798.CrossRefPubMed
39.
Mitra SK, Hanson DA, Schlaepfer DD: Focal adhesion kinase: in command and control of cell motility. Nat Rev Mol Cell Biol. 2005, 6: 56-68. 10.1038/nrm1549.CrossRefPubMed
40.
Akasaka T, van Leeuwen RL, Yoshinaga IG, Mihm MC, Byers HR: Focal adhesion kinase (p125FAK) expression correlates with motility of human melanoma cell lines. J Invest Dermatol. 1995, 105: 104-108. 10.1111/1523-1747.ep12313396.CrossRefPubMed
41.
Hess AR, Postovit LM, Margaryan NV, Seftor EA, Schneider GB, Seftor RE, Nickoloff BJ, Hendrix MJ: Focal adhesion kinase promotes the aggressive melanoma phenotype. Cancer Res. 2005, 65: 9851-9860. 10.1158/0008-5472.CAN-05-2172.CrossRefPubMed
42.
Bertoloucci CM, Guibao CD, Zheng J: Structural features of the focal adhesion kinase-paxillin complex give insight into the dynamics of focal adhesion assembly. Protein Sci. 2005, 14: 644-652. 10.1110/ps.041107205.CrossRef
43.
Cooley MA, Broome JM, Ohngemach C, Romer LH, Schaller MD: Paxillin binding is not the sole determinant of focal adhesion localization or dominant-negative activity of Focal Adhesion Kinase/Focal Adhesion Kinase-related Nonkinase. Mol Biol Cell. 2000, 11: 3247-3263.PubMedCentralCrossRefPubMed
44.
Shen Y, Schaller MD: Focal Adhesion Targeting: the critical determinant of FAK regulation and substrate phosphorylation. Mol Biol Cell. 1999, 10: 2507-2518.PubMedCentralCrossRefPubMed
45.
Hagel M, George EL, Kim A, Tamimi R, Opitz SL, Turner CE, Imamoto A, Thomas SM: The adaptor protein paxillin is essential for normal development in the mouse and is a critical transducer of fibronectin signaling. Mol Cell Biol. 2002, 22: 901-915.PubMedCentralCrossRefPubMed
46.
Hu YL, Haga JH, Miao H, Wang Y, Li YS, Chien S: Roles of microfilaments and microtubules in paxillin dynamics. Biochem Biophys Res Commun. 2006, 348: 1463-1471. 10.1016/j.bbrc.2006.08.022.CrossRefPubMed
47.
Schaller MD, Parsons JT: pp125FAK-dependent tyrosine phosphorylation of paxillin creates a high-affinity binding site for Crk. Mol Cell Biol. 1995, 15: 2635-2345.PubMedCentralCrossRefPubMed
48.
Kiyokawa E, Hashimoto Y, Kobayashi S, Sugimura H, Kurata T, Matsuda M: Activation of Rac1 by a Crk SH3-binding protein, DOCK180. Genes Dev. 1998, 12: 3331-3336. 10.1101/gad.12.21.3331.PubMedCentralCrossRefPubMed
49.
Webb DJ, Horwitz AF: New dimensions in cell migration. Nat Cell Biol. 2003, 5: 690-692. 10.1038/ncb0803-690.CrossRefPubMed
50.
Ferraro D, Corso S, Fasano E, Panieri E, Santangelo R, Borrello S, Giordano S, Pani G, Galeotti T: Pro-metastatic signaling by c-Met through RAC-1 and reactive oxygen species (ROS). Oncogene. 2006, 25: 3689-3698. 10.1038/sj.onc.1209409.CrossRefPubMed
51.
Kawaii S, Tomono Y, Ogawa K, Sugiura M, Yano M, Yoshizawa Y: The antiproliferative effect of coumarins on several cancer cell lines. Anticancer Res. 2001, 21: 917-923.PubMed
52.
Ratanasavanh D, Lamiable D, Biour M, Guédès Y, Gersberg M, Leutenegger E, Riché C: Metabolism and toxicity of coumarin on cultured human, rat, mouse and rabbit hepatocytes. Fundam Clin Pharmacol. 1996, 10: 504-510.CrossRefPubMed
53.
Nagata M, Fujita H, Ida H, Hoshina H, Inoue T, Seki Y, Ohnishi M, Ohyama T, Shingaki S, Kaji M, Saku T, Takagi R: Identification of potential biomarkers of lymph node metastasis in oral squamous cell carcinoma by cDNA microarray analysis. Int J Cancer. 2003, 106: 683-689. 10.1002/ijc.11283.CrossRefPubMed
54.
Li HG, Xie DR, Shen XM, Li HH, Zeng H, Zeng YJ: Clinicopathological significance of expression of paxillin, syndecan-1 and EMMPRIN in hepatocellular carcinoma. World J Gastroenterol. 2005, 11: 1445-14451.PubMedCentralCrossRefPubMed
55.
Vadlamudi R, Adam L, Tseng B, Costa L, Kumar R: Transcriptional up-regulation of paxillin expression by heregulin in human breast cancer cells. Cancer Res. 1999, 59: 2843-2846.PubMed
56.
Madan R, Smolkin MB, Cocker R, Fayyad R, Oktay MH: Focal adhesion proteins as markers of malignant transformation and prognostic indicators in breast carcinoma. Hum Pathol. 2006, 37: 9-15. 10.1016/j.humpath.2005.09.024.CrossRefPubMed
57.
Salgia R, Li JL, Ewaniuk DS, Wang YB, Sattler M, Chen WC, Richards W, Pisick E, Shapiro GI, Rollins BJ, Chen LB, Griffin JD, Sugarbaker DJ: Expression of the focal adhesion protein paxillin in lung cancer and its relation to cell motility. Oncogene. 1999, 18: 67-77. 10.1038/sj.onc.1202273.CrossRefPubMed
58.
Lamerant N, Kieda C: Adhesion properties of adhesion-regulating molecule 1 protein on endothelial cells. FEBS J. 2005, 272: 1833-1844. 10.1111/j.1742-4658.2005.04613.x.CrossRefPubMed
59.
Shimada S, Ogawa M, Takahashi M, Schlom J, Greiner W: Molecular cloning and characterization of the complementary DNA of an Mr 110,000 antigen expressed by human gastric carcinoma cells and upregulated by γ-Interferon. Cancer Res. 1994, 54: 3831-3836.PubMed
60.
Jørgensen JP, Lauridsen AM, Kristensen P, Dissing K, Johnsen AH, Hendil KB, Hartmann-Petersen R: Adrm1, a putative cell adhesion regulating protein, is a novel proteasome-associated factor. J Mol Biol. 2006, 360: 1043-1052. 10.1016/j.jmb.2006.06.011.CrossRefPubMed
Metadata
Title
Reduced paxillin expression contributes to the antimetastatic effect of 4-hydroxycoumarin on B16-F10 melanoma cells
Authors
Marco A Velasco-Velázquez
Nohemí Salinas-Jazmín
Nicandro Mendoza-Patiño
Juan J Mandoki
Publication date
01-12-2008
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2008
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/1475-2867-8-8

Other articles of this Issue 1/2008

Cancer Cell International 1/2008 Go to the issue

Primary research

Role of IGF-1/IGF-1R in regulation of invasion in DU145 prostate cancer cells

Primary research

Down-regulation of DNMT3b in PC3 cells effects locus-specific DNA methylation, and represses cellular growth and migration

Primary research

Ku86 exists as both a full-length and a protease-sensitive natural variant in multiple myeloma cells

Primary research

S100A4 overexpression proves to be independent marker for breast cancer progression

Primary research

Deregulation of histone lysine methyltransferases contributes to oncogenic transformation of human bronchoepithelial cells

Primary research

Nuclear localization and intensity of staining of nm23 protein is useful marker for breast cancer progression
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits