Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells. Issue 5 (5th January 2017)

• Record Type: Journal Article
• Authors: Das, Lipsa; Anderson, Todd A.; Gard, Jaime M.C.; Sroka, Isis C.; Strautman, Stephanie R.; Nagle, Raymond B.; Morrissey, Colm; Knudsen, Beatrice S.; Cress, Anne E.
• Is Part Of: Journal of cellular biochemistry. Volume 118:Issue 5(2017)
• Page Start: 1038
• Page End: 1049
• Publication Date: 2017-01-05
• Subjects: INTEGRIN -- LAMININ -- INTERNALIZATION KINETICS -- ENDOSOMES -- PROSTATE CANCER; Cytochemistry -- Periodicals; 572
• DOI: 10.1002/jcb.25673
• Languages: English
• ISSNs: 0730-2312

Abstract

ABSTRACT: Laminin binding integrins α6 (CD49f) and α3 (CD49c) are persistently but differentially expressed in prostate cancer (PCa). Integrin internalization is an important determinant of their cell surface expression and function. Using flow cytometry, and first order kinetic modeling, we quantitated the intrinsic internalization rates of integrin subunits in a single cycle of internalization. In PCa cell line DU145, α6 integrin internalized with a rate constant (kactual ) of 3.25 min −1, threefold faster than α3 integrin (1.0 min −1 ), 1.5‐fold faster than the vitronectin binding αv integrin (CD51) (2.2 min −1 ), and significantly slower than the unrelated transferrin receptor (CD71) (15 min −1 ). Silencing of α3 integrin protein expression in DU145, PC3, and PC3B1 cells resulted in up to a 1.71‐fold increase in kactual for α6 integrin. The internalized α6 integrin was targeted to early endosomes but not to lamp1 vesicles. Depletion of α3 integrin expression resulted in redistribution of α6β4 integrin to an observed cell–cell staining pattern that is consistent with a suprabasal distribution observed in epidermis and early PIN lesions in PCa. Depletion of α3 integrin increased cell migration by 1.8‐fold, which was dependent on α6β1 integrin. Silencing of α6 integrin expression however, had no significant effect on the kactual of α3 integrin or its distribution in early endosomes. These results indicate that α3 and α6 integrins have significantly different internalization kinetics and that coordination exists between them for internalization. J. Cell. Biochem. 118: 1038–1049, 2017. © 2016 Wiley Periodicals, Inc. Abstract : We developed a rate kinetic model of internalization and determined the basal internalization rates of laminin binding integrins (α3 and α6) to be significantly different. Coordination existed between their internalization such that, the loss of α3 integrin expression increased the internalization of α6 integrin to early endosomes and cell–cell locations. The results are important in the context of collective migration during epithelial tumor dissemination and metastasis using α6 and α3 integrins.