Abstract
Integrins play a major role in cell adhesion and migration. Previous work reported that a cleaved form of integrin α6 (α6p) was detected in invasive human prostate cancer tissue, absent in normal prostate tissue and was produced by urokinase-type Plasminogen Activator (uPA) in a plasmin-independent manner. Using site-directed mutagenesis we identified amino acid residues R594 and R595, located in the "stalk" region of integrin α6, as essential for cleavage. The cleavage site is located on the extracellular region of the protein between the β-barrel domain and the thigh domain. Prostate cancer cells (PC3N) were stably transfected to overexpress the cleavable, wild-type (PC3N-α6-WT) or the non-cleavable form of integrin α6 (PC3N-α6-RR). The number of cells invading laminin 111- and laminin 332-coated filters by PC3N-α6-WT cells increased by threefold as compared to PC3N-α6-RR cells. Plasminogen activator inhibitor-1 (PAI-1) reduced the invasion of PC3N-α6-WT cells by approximately 42% through laminin 332-coated filters and plasmin inhibitor aprotinin had no significant effect. Linear cell migration increased production of integrin α6p in the PC3N-α6-WT cells and not in the PC3N-α6-RR cells and 32% of the PC3N-α6-WT cells migrated on laminin 111 in the linear migration assay as compared to the 5% PC3N-α6-RR cells. These data taken together suggest that the uPA-mediated cell surface cleavage of the α6 integrin extracellular domain is involved in tumor cell invasion and migration on laminin.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1080-1089 |
| Number of pages | 10 |
| Journal | Experimental Cell Research |
| Volume | 313 |
| Issue number | 6 |
| DOIs | |
| State | Published - Apr 1 2007 |
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Keywords
- Cleavage
- ECM
- Integrin α6
- Laminin
- Migration
- Prostate cancer
- uPA
- Urokinase
ASJC Scopus subject areas
- Cell Biology
Cite this
Integrin α6 cleavage : A novel modification to modulate cell migration. / Pawar, Sangita C.; Demetriou, Manolis C.; Nagle, Raymond B; Bowden, G. Tim; Cress, Anne E.
In: Experimental Cell Research, Vol. 313, No. 6, 01.04.2007, p. 1080-1089.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Integrin α6 cleavage
T2 - A novel modification to modulate cell migration
AU - Pawar, Sangita C.
AU - Demetriou, Manolis C.
AU - Nagle, Raymond B
AU - Bowden, G. Tim
AU - Cress, Anne E
PY - 2007/4/1
Y1 - 2007/4/1
N2 - Integrins play a major role in cell adhesion and migration. Previous work reported that a cleaved form of integrin α6 (α6p) was detected in invasive human prostate cancer tissue, absent in normal prostate tissue and was produced by urokinase-type Plasminogen Activator (uPA) in a plasmin-independent manner. Using site-directed mutagenesis we identified amino acid residues R594 and R595, located in the "stalk" region of integrin α6, as essential for cleavage. The cleavage site is located on the extracellular region of the protein between the β-barrel domain and the thigh domain. Prostate cancer cells (PC3N) were stably transfected to overexpress the cleavable, wild-type (PC3N-α6-WT) or the non-cleavable form of integrin α6 (PC3N-α6-RR). The number of cells invading laminin 111- and laminin 332-coated filters by PC3N-α6-WT cells increased by threefold as compared to PC3N-α6-RR cells. Plasminogen activator inhibitor-1 (PAI-1) reduced the invasion of PC3N-α6-WT cells by approximately 42% through laminin 332-coated filters and plasmin inhibitor aprotinin had no significant effect. Linear cell migration increased production of integrin α6p in the PC3N-α6-WT cells and not in the PC3N-α6-RR cells and 32% of the PC3N-α6-WT cells migrated on laminin 111 in the linear migration assay as compared to the 5% PC3N-α6-RR cells. These data taken together suggest that the uPA-mediated cell surface cleavage of the α6 integrin extracellular domain is involved in tumor cell invasion and migration on laminin.
AB - Integrins play a major role in cell adhesion and migration. Previous work reported that a cleaved form of integrin α6 (α6p) was detected in invasive human prostate cancer tissue, absent in normal prostate tissue and was produced by urokinase-type Plasminogen Activator (uPA) in a plasmin-independent manner. Using site-directed mutagenesis we identified amino acid residues R594 and R595, located in the "stalk" region of integrin α6, as essential for cleavage. The cleavage site is located on the extracellular region of the protein between the β-barrel domain and the thigh domain. Prostate cancer cells (PC3N) were stably transfected to overexpress the cleavable, wild-type (PC3N-α6-WT) or the non-cleavable form of integrin α6 (PC3N-α6-RR). The number of cells invading laminin 111- and laminin 332-coated filters by PC3N-α6-WT cells increased by threefold as compared to PC3N-α6-RR cells. Plasminogen activator inhibitor-1 (PAI-1) reduced the invasion of PC3N-α6-WT cells by approximately 42% through laminin 332-coated filters and plasmin inhibitor aprotinin had no significant effect. Linear cell migration increased production of integrin α6p in the PC3N-α6-WT cells and not in the PC3N-α6-RR cells and 32% of the PC3N-α6-WT cells migrated on laminin 111 in the linear migration assay as compared to the 5% PC3N-α6-RR cells. These data taken together suggest that the uPA-mediated cell surface cleavage of the α6 integrin extracellular domain is involved in tumor cell invasion and migration on laminin.
KW - Cleavage
KW - ECM
KW - Integrin α6
KW - Laminin
KW - Migration
KW - Prostate cancer
KW - uPA
KW - Urokinase
UR - http://www.scopus.com/inward/record.url?scp=33947261720&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33947261720&partnerID=8YFLogxK
U2 - 10.1016/j.yexcr.2007.01.006
DO - 10.1016/j.yexcr.2007.01.006
M3 - Article
C2 - 17303120
AN - SCOPUS:33947261720
VL - 313
SP - 1080
EP - 1089
JO - Experimental Cell Research
JF - Experimental Cell Research
SN - 0014-4827
IS - 6
ER -