Highly invasive cell subpopulations from a human prostate carcinoma cell line, PC-3, were selected for by allowing the parental PC-3 cells to invade through reconstituted basement membrane, Matrigel. These cells were collected, cultured and then selected further by repeated invasion through the in vitro invasion chamber. The invasive subpopulations (I-PC3 (2) and (3)) were found to be approximately 15-fold more invasive in vitro than the parental cells, had a distinct rounded morphology in culture, and proliferated more rapidly than the parental cells. When injected either subcutaneously or intraperitoneally into immunocompromised SCID mice, the I-PC3 cells were found to form tumors at the primary sites and to be highly invasive and metastatic. In contrast, the parental PC-3 cells formed tumors at the site of inoculation in these mice but failed to invade or metastasize. The I-PC3 cells attached equally as well as PC-3 cells to fibronectin, laminin, collagen type IV and vitronectin, but unlike the parental PC-3 cells these invasive variants failed to spread on any of these substrates. On Matrigel, the PC-3 cells became highly organized, whereas the I-PC3 cells remained rounded, clumped together and penetrated into the Matrigel. Biochemical analysis of the expression of adhesion proteins and integrins demonstrated that whereas the parental cells synthesized and secreted substantial amounts of fibronectin, the I-PC3 cell variants did not secrete any fibronectin. Although both PC-3 and I-PC3 cells expressed equivalent levels of cell surface αvβ 3, α2β1 and α5β1 integrins, the expression of the α3β1 integrin, which is expressed at very high levels on the parental PC-3 cells, was drastically reduced on the invasive I-PC3 cells. This decrease in expression of α3 occurred also at the level of mRNA expression. Finally, whereas the PC-3 cells express α6β1, in the invasive I-PC3 cells the a6 subunit was associated mostly with the β4 subunit. Since the α6β4 integrin is analogous to the A9 tumor antigen which is associated with aggressive human squamous cell carcinomas, the apparent overexpression of α6β4 may also participate in the aggressive behavior of these variant prostate carcinoma cells. Alterations in the expression of the α3β1 and α6β4 integrins may thus allow these cells to become more invasive, and lead to an increased propensity for metastasis.
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References
Liotta LA, Rao CN and Weiver UM, 1986, Biochemical interactions of tumor cells with the basement membrane. Annual Review in Biochemistry, 55, 1037–1057.
Plantefaber LC and Hynes RO, 1989, Changes in integrin receptors on oncogenically transformed cells. Cell, 56, 281–290.
Hynes RO. Surfaces of Normal and Malignant Cells. New York: John Wiley, 1979.
Hynes RO and Yamada KM, 1982, Fibronectins: multifunctional modular glycoproteins. Journal of Cell Biology, 95, 369–377.
Dedher S, 1990, Integrins and tumor invasion. BioEssays, 12, 583–590.
Hynes RO, 1992, Integrins: versatility, modulation, and signaling in cell adhesion. Cell, 69, 11–25.
Giancotti FG and Ruoslahti E, 1990, Elevated levels of the α5β1 fibronectin receptor suppresses the transformed phenotype of Chinese hamster ovary cells. Cell, 60, 849–859.
Dedhar S, Argraves WS, Suzuki R, Ruoslahti E and Pierschbacher MD, 1987, Human osterosarcoma cells resistance to detachment by an Arg-Gly-Asp containing peptide overproduce the fibronectin receptor. Jounal of Cell Biology, 105, 1175–1182.
Albeida SM, Mette SA, Elder DE, Stewart R, Danjanovich K, Herlyn M and Buck CA, 1990, Integrin distribution in malignant melanomas: association of the β3 subunit with tumor progression. Cancer Research, 50, 6757–6764.
Felding-Habermann B, Mueller BM, Romerdahl CA and Cheresh D, 1992, Involvement of αv gene expression in human melanoma tumorigenesis. Journal of Clinical Investigation, 89, 2018.
Chan BM, Matsaura N, Takada Y, Zetter BR and Hemler ME, 1991, In vitro and in vivo consequences of VLA-2 expression on rhabdamyosarcoma cells. Science, 251, 1600–1602.
Pignatelli M, Harby AM and Stamp GWH, 1991, Low expression of β1, α2 and α3 subunits of VLA integrins in malignant mammary tumors. Journal of Pathology, 165, 25–32.
Pignatelli M, Smith MEF and Bodmer WF, 1990, Low expression of collagen receptors in moderate and poorly differentiated colorectal adenocarcinomas. British Journal of Cancer, 61, 636–638.
Van Waes C, Kozasky KF, Warren AB, Kidd L, Paugh D, Liebert M and Carey TE, 1991, The A9 antigen associated with aggressive human squamous carcinoma is structurally and functionally similar to the newly defined integrin α6β4. Cancer Research, 51, 2395–2402.
Falcioni R, Kennel SJ, Giacomini P, Zupi G and Sacchi A, 1986, Expression of tumor antigen correlated with metastatic potential of Lewis lung carcinoma and B16 melanoma clones in mice. Cancer Research, 46, 5772–5778.
Kennel SJ, Foote LJ, Falcioni R, Sonnenberg A, Stringer CD, Crouse C and Hemler ME, 1989, Analysis of the tumor-associated antigen TSP-180. Identify with α6β4 in the integrin superfamily. Journal of Biological Chemistry, 264, 15515–15521.
Takada Y, Murphy E, Pil P, Chen C, Ginsberg MH and Hemler ME, 1991, Molecular cloning and expression of the cDNA for α3 subunit of human α3β1 (VLA-3), an integrin receptor for fibronectin, laminin and collagen. Journal of Cell Biology, 115, 257–266.
Dedhar S, Haqq C and Gray V, 1989, Specific overproduction of VLA-1 integrin in two human neuroblastoma cell lines selected for resistance to detachment by an Arg-Gly-Asp containing synthetic peptide. Journal of Biological Chemistry, 264, 4832–4836.
Laemmli UK, 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.
Overall CM, Wrana JL and Sodek J, 1989, Independent regulation of collagenase 72 kDa progelatinase and metalloproteinase inhibitor expression in human fibroblasts by transforming growth factor β. Journal of Biological Chemistry, 264, 1860–1869.
Overall CM and Sodek JJ, 1990, Concanavalin A produces a matrix degrative phenotype in human fibroblasts. Journal of Biological Chemistry, 265, 21141–21151.
Maniatis T, Fritsch EF and Sambrook J. Molecular Cloning: a laboratory manual. New York: Cold Spring Harbor Laboratory, 1982.
Ullrich AP, Shine J, Chirgwin J, Pictet R, Tischer E, Rutter WJ and Goodman HM, 1977, Rat insulin genes: construction of plasmids containing the coding sequences. Science, 196, 1313–1315.
Ruoslahti E, Hayman EG, Pierschbacher MD and Engvall E, 1982, Fibronectin: purification, immunochemical properties and biological activities. Methods of Enzymology, 82, 803–831.
Dedhar S and Saulnier R, 1990, Alterations in integrin receptor expression on chemically transformed human cells: specific enhancement of laminin and collagen receptor complexes. Journal of Cell Biology, 110, 481–489.
Kaighn ME, Lechner JF, Narayan KS and Jones LW, 1978, Prostate carcinoma: tissue culture cell lines. National Cancer Institute Monograph, 49, 17–21.
Kornberg L, Earp HS, Parsons JT, Schaller M and Juliano RL, 1992, Cell adhesion of integrin clustering increases phosphorylation of a focal adhesion associated tyrosine kinase. Journal of Biological Chemistry, 267, 23439–23442.
Guan JL and Shalloway D, 1992, Regulation of focal adhesion associated protein tyrosine kinase by both cellular adhesion and oncogene transformation. Nature, 358, 390–392.
Dedhar S, Jewell K, Rojiani M and Gray V, 1992, The receptor for the basement membrane glycoprotein entactin is the integrin α3/β1. Journal of Biological Chemistry, 267, 18908–18914.
Carey TF, Wolf GT, Hsu S, Poore J, Peterson K and McCatchey KD, 1987, Expression of A9 antigen and loss of blood group antigens as determinants of survival in patients with head and neck squamous carcinoma. Otolarynology Head and Neck Surgery, 96, 221–230.
Wolf GT, Carey TE, Schmaltz SP, McCatchey KD, Poore J, Glaser L, Hayashida DJS and Hsu S, 1990, Altered antigen expression predicts outcome in squamous cell carcinoma of the hand and neck. Journal of the National Cancer Institute, 82, 1566–1572.
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Dedhar, S., Saulnier, R., Nagle, R. et al. Specific alterations in the expression of α3β1 and α6β4 integrins in highly invasive and metastatic variants of human prostate carcinoma cells selected by in vitro invasion through reconstituted basement membrane. Clin Exp Metast 11, 391–400 (1993). https://doi.org/10.1007/BF00132982
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DOI: https://doi.org/10.1007/BF00132982