Inhibition of lysophosphatidic acid receptor-2 expression by RNA interference decreases lysophosphatidic acid-induced urokinase plasminogen activator activation, cell invasion, and migration in ovarian cancer SKOV-3 cells

To explore the role of lysophosphatidic acid receptor-2 (LPA2) in regulating lysophosphatidic acid (LPA)-induced urokinase plasminogen activator (uPA) activation, cell invasion, and migration in human ovarian cancer cell line SKOV-3. SKOV-3 cells were stimulated with LPA. Cell supernatant uPA level...

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Published in:Croatian medical journal 2008-04, Vol.49 (2), p.175-181
Main Authors: Wang, Gui-li, Wen, Ze-qing, Xu, Wan-peng, Wang, Zhi-yu, Du, Xue-lian, Wang, Fei
Format: Article
Language:English
Subjects:
Basic Science
Cell Growth Processes
Cell Line, Tumor
Cell Movement
Cell receptors
Chemotaxis
Enzyme-Linked Immunosorbent Assay
Female
Humans
In Vitro Techniques
K562 Cells
Lysophospholipids - pharmacology
Ovarian cancer
Ovarian Neoplasms - physiopathology
Pilot Projects
Receptors, Lysophosphatidic Acid - antagonists & inhibitors
RNA Interference
RNA, Messenger
Signal Transduction
Urokinase
Urokinase-Type Plasminogen Activator - biosynthesis
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Summary:To explore the role of lysophosphatidic acid receptor-2 (LPA2) in regulating lysophosphatidic acid (LPA)-induced urokinase plasminogen activator (uPA) activation, cell invasion, and migration in human ovarian cancer cell line SKOV-3. SKOV-3 cells were stimulated with LPA. Cell supernatant uPA level and activity were measured using enzyme-linked immunosorbent assay. LPA2 mRNA expression was inhibited with LPA2-specific small interfering RNA (siRNA) and examined using semiquantitative reverse transcriptase-polymerase chain reaction. LPA-induced cell invasion and migration in transfected cells were evaluated by a Matrigel invasion chamber and a Transwell chemotaxis chamber, respectively. LPA stimulation significantly enhanced in vitro uPA activity in time- and dose-dependent manner. The levels of LPA-induced uPA protein decreased by 55% in LPA2 siRNA-transfected cells compared with negatively transfected cells at 24 hours after being treated with 80 micromol/L LPA (0.75+/-0.03 vs 0.34+/-0.04, P=0.004). In the LPA2 specific siRNA-transfected SKOV-3 cells, LPA treatment at 80 micromol/L induced considerably less invasion and migration compared with negative control siRNA-transfected SKOV-3 cells (invasion: 178+/-17.2 vs 36.2+/-3.3, P=0.009; migration: 220.4+/-25.5 vs 57+/-7.6, P=0.009). LPA2 has an essential role in LPA-induced uPA activation and tumor cell invasion in ovarian cancer SKOV-3 cells.
ISSN:0353-9504
1332-8166
DOI:10.3325/cmj.2008.2.175