Abstract
Cellular adhesion to extracellular matrix proteins via integrin molecules is a major factor in the process of invasion and metastasis of human tumor cells. Four human prostate cell lines were characterized according to the presence and quantity of integrin subunits, the ability of the cells to attach to extracellular substrates and the capacity of the cells to form tumors in severe combined immunodeficient (SCID) mice. All four human prostate cell lines expressed three to five integrins on their cell surfaces. The DU145, PC3 and 431P cells expressed primarily α3, α5, and α6 integrin at similar levels. These cell lines expressed the subunits β1, β3 and β4 with β1 predominant. The DU145 cells preferred attachment to fibronectin, followed by laminin and vitronectin. Approximately 50%–60% of the binding of DU145 cells to fibronectin and laminin was dependent on the function of α5β1 and α6 respectively. The cell line LNCaP differed in its low expression of the α3 subunit, 95% of cellular adhesion to fobronectin and laminin being integrin-dependent and its inability to attach to vitronectin, in spite of surface expression of αvβ3. All the cell lines except for LNCaP readily formed tumors within SCID mice and the expression of α3, α6, β1, and β4 integrin subunits was preserved in the resulting tumor tissue. The altered adhesion properties of the LNCaP cells may explain their altered tumorigenicity.
Key words: Integrins, Cell adhesion, Human prostate cell lines, Metastasis
Abbreviations
- SCID
severe combined immunodeficiency
- FITC
fluorescein isothiocynate
- FACS
fluorescence-activated cell sorting
- PBS
phosphate-buffered saline
- FBS
fetal bovine serum
Footnotes
This work was supported in part by a grant from the Friends of the Arizona Cancer Center, Phoenix Chapter, is ACS grant PDT-388 and CH-467 and CA 56666
References
- Albelda SM, Buck CA (1990) Integrins and other cell adhesion molecules. Integrin Review. FASEB J 4:2868–2880 [PubMed] [Google Scholar]
- Albelda SM, Mette SA, Elder AE, Stewart RM, Damjanovich L, Herlyn M, Buck CA (1990) Integrin distribution in malignant melanoma: association of the β3 subunit with tumor progression. Cancer Res 50:6757–6764 [PubMed] [Google Scholar]
- Ansell JD, Bancroft GJ (1989) The biology of the SCID mutation. Immunol Today 10:322–325 [DOI] [PubMed] [Google Scholar]
- Bauer JS, Schreiner CL, Giancotti FG, Ruoslahti E, Juliano RL (1992) Motility of fibronectin receptor-deficient cells on fibronectin and vitronectin: collaborative interactions among integrins. J Cell Biol 116:477–487 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bosco M, Chan C, Matsuura N, Takada Y, Zetter BR, Hemler ME (1991) In vitro and in vivo consequences of VLA-2 expression on rhabdomyosarcoma cells. Science 251:1600–1602 [DOI] [PubMed] [Google Scholar]
- Bosma MJ, Carroll AM (1991) The SCID mouse mutant: definition, characterization and potential uses. Annu Rev Immunol 9:323–350 [DOI] [PubMed] [Google Scholar]
- Burridge K (1986) Substrate adhesions in normal and transformed fibroblasts: organization and regulation of cytoskeletal, membrane and extracellular matrix components at focal contacts. Cancer Res 4:18–78 [Google Scholar]
- Burridge K, Fath K, Kelly T, Nuckolis B, Turner C (1988) Focal adhesions: transmembrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol 4:487–525 [DOI] [PubMed] [Google Scholar]
- Chan R, Rossito PV, Edwards BF, Cardiff RD (1986) Presence of proteolytically processed keratins in culture medium of MCF-7. Cancer Res 46:6353–6359 [PubMed] [Google Scholar]
- Chelberg MK, Tsilibrary EC, Hauser AR, McCarthy JB (1989) Type IV collagen-medicated melanoma cell adhesion and migration: involvement of multiple, distinct domains of the collagen molecule. Cancer Res 49:4796–4802 [PubMed] [Google Scholar]
- Cheresh DA, Smith JW, Cooper HM, Quaranta V (1989) A novel vitronectin receptor integrin (αvβx) is responsible for distinct adhesive properties of carcinoma cells. Cell 57:59–69 [DOI] [PubMed] [Google Scholar]
- Chung LWK (1991) Fibroblasts are critical determinants in prostatic cancer growth and dissemination. Cancer Metastasis Rev 10:263–274 [DOI] [PubMed] [Google Scholar]
- Chung LWK, Gleave ME, Hsieh, JT, Hong SJ, Zhau HE (1991) Reciprocal mesenchymal-epithelial interaction affecting prostate tumor growth and hormonal responsiveness. Cancer Surv 11:91–121 [PubMed] [Google Scholar]
- Costantini RM, Falcioni R, Battista P, Zupi G, Kennel SJ, Colasante A, Venturo I, Curcio CG, Sacchi A (1990) Integrin (α6/β4) expression in human lung cancer as monitored by specific monoclonal antibodies. Cancer Res 50:6107–6112 [PubMed] [Google Scholar]
- Dedhar S, Saulnier R (1990) Alterations in integrin receptor expression on chemically transformed human cells: specific enhancement of laminin and collagen receptor complexes. J Cell Biol 110:481–489 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Du XP, Plow EF, III Frelinger AL, O'Toole TE, Loftus JC, Ginsberg MH (1991) Ligands “activate” integrin α11bβ3 (platelet GPIIb-IIIa). Cell 65:409–416 [DOI] [PubMed] [Google Scholar]
- Feldman LE, Shin KC, Natale RB, Todd RF (1991) 643-5 integrin expression on human small cell lung cancer cells. Cancer Res 51:1065–1070 [PubMed] [Google Scholar]
- Freed E, Gailit J, van der Geer P, Ruoslahti E, Hunter T (1989) A novel integrin β subunit is associated with the vitronectin receptor a subunit (αv) in a human osteosarcoma cell line and is a substrate for protein kinase C. EMBO J 8:2955–2965 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Freeman MR, Song Y, Carson DD, Guthrie PD, Chung LWK (1991) Extracellular matrix and androgen receptor expression associated with spontaneous transformation of rat prostate fibroblasts. Cancer Res 51:1910–1916 [PubMed] [Google Scholar]
- Gleave M, Hsieh JT, Gao C, von Eschenbach AC, Chung LWK (1991) Acceleration of humanprostate cancer growth in vivo by factors produced by prostate and bone fibroblasts. Cancer Res 51:3753–3761 [PubMed] [Google Scholar]
- Gleave ME, Hsieh JT, von Eschenbach AC, Chung LWK (1992) Prostate and bone fibroblasts induce human prostate cancer growth in vivo: implications for bidirectional tumor-stromal cell interaction in prostate carcinoma growth and metastasis. J Urol 147:1151–1159 [DOI] [PubMed] [Google Scholar]
- Hemler ME, Ware CF, Strominger JL (1983) Characterization of a novel differentiation antigen complex recognized by a monoclonal antibody (A-1A5): unique activation-specific molecular forms on stimulated T cells. J Immunol 131:334–340 [PubMed] [Google Scholar]
- Hessle H, Sakai L, Hollister D, Burgeson R, Engvall E (1984) Basement membrane diversity detected by monoclonal antibodies. Differentiation 26:49–54 [DOI] [PubMed] [Google Scholar]
- Hogervorst F, Kuikman I, von dem Borne AEG, Sonnenberg A (1990) Cloning and sequence analysis of beta-4 cDNA: an integrin subunit that contains a unique 118 kd cytoplasmic domain. EMBO J 9:765–770 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horoszewicz JS, Leong SS, Kawinski E, Karr JP, Rosenthal H, Chu TM, Mirand EA, Murphy GP (1983) LNCaP model of human prostatic carcinoma. Cancer Res 43:1809–1818 [PubMed] [Google Scholar]
- Hynes RO (1992) Integrins: versitility, modulation and signaling in cell adhesion. Cell 69:11–25 [DOI] [PubMed] [Google Scholar]
- Isberg RR, Leong JM (1990) Multiple α1 chain integrins are receptors for invasin a protein that promotes bacterial penetration into mammalian cells. Cell 69 861–871 [DOI] [PubMed] [Google Scholar]
- Kaighn ME, Narayan KS, Ohnuki Y, Lechner JF, Jones LW (1979) Establishment and characterization of a human prostate carcinoma cell line (PC-3). Invest Urol 17:16–23 [PubMed] [Google Scholar]
- Kramer RH, Vu M, Cheng Y-F, Ramos DM (1991) Integrin expression in malignant melanoma. Cancer Metastasis Rev 10:49–59 [DOI] [PubMed] [Google Scholar]
- La Flamme SE, Akiyama SK, Yamada KM (1992) Regulation of fibronectin receptor distribution. J Cell Biol 117:437–447 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee EC, Lotz MM, Steele GD Jr, Mercurio AM (1992) The integrin α6β4 is a laminin receptor. J Cell Biol 117:671–678 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liotta LA, Kohn E (1990) Cancer invasion and metastases. JAMA 263:1123–1126 [PubMed] [Google Scholar]
- Long MM, King VJ, Prasad KU, Freeman BA, Urry DW (1989) Elastin repeat peptide as chemoattractants for bovine aortic endothelial cells. J Cell Physiol 140:512–518 [DOI] [PubMed] [Google Scholar]
- Passaniti A, Issacs JT, Haney JA, Adler SW, Cujdik TJ, Long PV, Kleinman HK (1992) Stimulation of human prostatic carcinoma tumor growth in athymic mice and control of migration in culture be extracellular matrix. Int J Cancer 51:318–324 [DOI] [PubMed] [Google Scholar]
- Plantefaber LC, Hynes RO (1989) Changes in integrin receptors on oncogenically transformed cells. Cell 56:281–290 [DOI] [PubMed] [Google Scholar]
- Pretlow TG, Delmoro CM, Dilley GG, Spadafora CG, Pretlow TP (1991) Transplantation of human prostatic carcinoma into nude mice in matrigel. Cancer Res 51:3814–3817 [PubMed] [Google Scholar]
- Ruoslahti E, Giancotti FG (1989) Integrins and tumor cell dissemination. Cancer Cells 1:119–126 [PubMed] [Google Scholar]
- Shevrin DH, Kikreja SC, Vincic B, Lad TE (1986) Patterns of spontaneous metastasis by human prostate cancer grown in athymic nude mice. Proc Am Assoc Cancer Res 27:256A [Google Scholar]
- Shevrin DH, Kukreja SC, Ghosh L, Lad TE (1988) Development of skeletal metastasis by human prostate cancer in athymic nude mice. Clin Exp Metastasis 6:401–409 [DOI] [PubMed] [Google Scholar]
- Smith JW, Vestal DJ, Irwin SV, Burke TA, Cheresh DA (1990) Purification and functional characterization of integrin αvβ5. J Biol Chem 265:11008–11013 [PubMed] [Google Scholar]
- Sonnenberg A, Janssen H, Hogervorst V, Calafat J, Hilgers J (1987) A complex of platelet glycoproteins Ic and IIa identified by a rat monoclonal antibody. J Biol Chem 262:10376–10383 [PubMed] [Google Scholar]
- Sonnenberg A, Moddermann PW, Hogervorst F (1988) Laminin receptor on platelets is the integrin VLA-6. Nature 336:487–489 [DOI] [PubMed] [Google Scholar]
- Stone KR, Mickey DD, Wunderli H, Mickey GH, Paulson DF (1978) Isolation of a human prostate carcinoma cell line (DU145). Int J Cancer 21:274–281 [DOI] [PubMed] [Google Scholar]
- Vogel BE, Tarone G, Giancotti FG, Gailit J, Ruoslahti E (1990) A novel fibronectin receptor with an unexpected subunit composition (αvβ1). J Biol Chem 265:5934–5937 [PubMed] [Google Scholar]
- Wayner EA, Carter WG, Piotrowicz RS, Kunicki TJ (1988) The function of multiple extracellular matrix receptors in mediating cell adhesion to extracellular matrix: preparation of monoclonal antibodies to the fibronectin receptor that specifically inhibit cell adhesion to fibronectin and react with platelet glycoproteins Ic-IIa. J Cell Biol 107:1881–1891 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wayner EA, Garcia-Pardo A, Humphries MJ, McDonald JA, Carter WG (1989) Identification and characterization of the T lymphocyte adhesion receptor for an alternative cell attachment domain (CS-1) in plasma fibronectin. J Cell Biol 69 1321–1330 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wayner EA, Orlando RA, Cheresh DA (1991) Integrins αvβ3 and αvβ5 contribute to cell attachment to vitronectin but differentially distribute on the cell surface. J Cell Biol 113:919–929 [DOI] [PMC free article] [PubMed] [Google Scholar]
- Young IT (1977) Proof without prejudice: use of the Kolmogorov-Smirnov test for the analysis of histograms from flow systems and other sources. J Histochem Cytochem 25:935–941 [DOI] [PubMed] [Google Scholar]
