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Objective : To study the ERK1 / 2 signaling inhibitor PD98059 on MGC-803 gastric cancer cells downstream of RECK and MMP-9 gene expression , and to explore ERK1 / 2 signaling pathway in RECK gene inhibits invasion and metastasis . Methods: in the logarithmic phase of human gastric cancer MGC-803 cells after synchronization packet that PD98059 at different concentrations (5,10,25,50,100 μmol / L) function 24h, and PD98059 25μmol / L effect at different times ( 2,4,12,24,48 h), and no drug-treated group as a control group , the DMSO (25μmol / L) as a negative control group , by: ( 1 ) MTT assay (MTT) was observed cancer cells MGC-803 proliferation . Select the appropriate drug concentration for further experiments . ( 2 ) protein research (Western-blot) assay P-ERK protein expression that ERK phosphorylation as a measure of ERK1 / 2 signaling pathway activity indicators . ( 3 ) semi-quantitative reverse transcription - polymerase chain reaction (RT-PCR) method and Western-blot assay RECK and MMP-9 mRNA and protein expression . Results : (1) MTT assay showed : PD98059 can inhibit cancer cell proliferation MGC803 , and in the role of 12, 24 and 48h time point was time-dependent (25μmol / L), at 25, 50 and 100μmol / L concentration point concentration- effect dependence (24h). The effective concentration of PD98059 is 25μmol / L, at this concentration valid for 24h. (2) Western-blot assay showed : PD98059 to inhibit ERK1 / 2 phosphorylation , the lower P-ERK1 / 2, so that the ERK1 / 2 signaling pathway inactivation. (3) RT-PCR method and Western-blot assay showed : PD98059 to concentration-dependent (25-100μmol / L) and time-dependent (12-48h) stimulation of cultured cancer cells RECK mRNA and protein expression , MMP -9mRNA and protein expression decreased . Conclusions : (1) PD98059 can inhibit human gastric cancer MGC-803 cells proliferation , growth . (2) PD98059 could upregulate the expression of RECK , reduced expression of MMP-9 , can be seen inhibiting ERK1 / 2 signaling pathway can inhibit the invasion and metastasis of human gastric cancer cells .
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