Effect Of Exogenous Osteopontin On The Proliferation, Invasiveness And Correlative Gene Expression Of Human Colorectal Carcinoma LS-174T Cells

Purpose: To explore the functional role of osteopontin (OPN) in colon cancer development and progression by investigating the effects of recombinant human osteopontin (rhOPN) on the proliferation ,invasiveness and mRNA transcription levels of matrix metallproteinase-2 (MMP-2), Vascular endothelial growth factor (VEGF) and urokinase type plasminogen activator (uPA) in human colorectal carcinoma LS-174T cells in vitro.Methods: After the LS-174T cells were treated with rhOPN in nmol/L at different concentrations for 24h or 20h, 3-(4,5-dimerthylthiazol-2-yl)-2,5-diphenylt tetrazolium bromide (MTT) colorimetric assay was used to examine cell proliferation, reconstituted basement membrane invasion assay was utilized to evaluate the invasive capacity, and real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to determine transcription levels of cellular MMP-2, VEGF and uPA mRNA. Relative quantifications of target genes expression, normalized by GAPDH housekeeping gene, were achieved by the way of the 2(-Delta Delta C(T)) method.Results:1. After the LS-174T cells were treated with rhOPN in nmol/L at different concentrations for 24h, all the MTT values increased (P<0.01) with obvious dosage dependent pattern. 2. When rhOPN at the concentrations of 5nmol/L or 30nmol/L was added to the upper chamber of transwell, the number of invaded cells was obviously more than (P<0.05) those of control group with dosage dependent pattern. 3. After the LS-174T cells were treated with different concentrations of rhOPN (5nomol/l,30nmol/L) for 24h, the transcription levels of cellular VEGF and uPA mRNA in LS-174T cells increased significantly (P<0.05, P<0.01) ,but the levels of...