Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells

Lipsa Das, Todd A. Anderson, Jaime M C Gard, Isis C Sroka, Stephanie R. Strautman, Raymond B Nagle, Colm Morrissey, Beatrice S. Knudsen, Anne E Cress

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1038-1049
Number of pages12
JournalJournal of Cellular Biochemistry
Volume118
Issue number5
DOIs
StatePublished - May 1 2017

Fingerprint

Laminin
Integrins
Prostatic Neoplasms
Cells
Endosomes
Vitronectin
Kinetics
Transferrin Receptors
Flow cytometry
Epidermis
Cell Movement

Keywords

  • ENDOSOMES
  • INTEGRIN
  • INTERNALIZATION KINETICS
  • LAMININ
  • PROSTATE CANCER

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells. / 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.

In: Journal of Cellular Biochemistry, Vol. 118, No. 5, 01.05.2017, p. 1038-1049.

Research output: Contribution to journalArticle

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. / Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells. In: Journal of Cellular Biochemistry. 2017 ; Vol. 118, No. 5. pp. 1038-1049.
@article{cf75aa829ba24724b5b50d1beb20b8bf,
title = "Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells",
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.",
keywords = "ENDOSOMES, INTEGRIN, INTERNALIZATION KINETICS, LAMININ, PROSTATE CANCER",
author = "Lipsa Das and Anderson, {Todd A.} and Gard, {Jaime M C} and Sroka, {Isis C} and Strautman, {Stephanie R.} and Nagle, {Raymond B} and Colm Morrissey and Knudsen, {Beatrice S.} and Cress, {Anne E}",
year = "2017",
month = "5",
day = "1",
doi = "10.1002/jcb.25673",
language = "English (US)",
volume = "118",
pages = "1038--1049",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "5",

}

TY - JOUR

T1 - Characterization of Laminin Binding Integrin Internalization in Prostate Cancer Cells

AU - Das, Lipsa

AU - Anderson, Todd A.

AU - Gard, Jaime M C

AU - Sroka, Isis C

AU - Strautman, Stephanie R.

AU - Nagle, Raymond B

AU - Morrissey, Colm

AU - Knudsen, Beatrice S.

AU - Cress, Anne E

PY - 2017/5/1

Y1 - 2017/5/1

N2 - 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.

AB - 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.

KW - ENDOSOMES

KW - INTEGRIN

KW - INTERNALIZATION KINETICS

KW - LAMININ

KW - PROSTATE CANCER

UR - http://www.scopus.com/inward/record.url?scp=85009188499&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85009188499&partnerID=8YFLogxK

U2 - 10.1002/jcb.25673

DO - 10.1002/jcb.25673

M3 - Article

C2 - 27509031

AN - SCOPUS:85009188499

VL - 118

SP - 1038

EP - 1049

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 5

ER -