Celecoxib For Non-small Cell Lung Cancer A549 Cell Proliferation, Invasion And The Influence Of The Adhesion And Mechanism Research

ObjectiveTo study the effects of celecoxib on growth and invasion in non-small cell lungcancer cell line A549.Methods（1） Cell growth assayThe A549cells in the logarithmic growth phase were randomly divided into onecontrol group （group C） and four treatment groups （group I₁、group I₂、groupI₃andgroup I₄）. The cells in group C were cultured commonly for48h. The cells intreatment groups were exposed on10ng/mL、20ng/mL、40ng/mL and80ng/mLIGF-1respectively for48h. The cell proliferation of each group was detected usingMTT assay. And the results showed the minimum effective dose of IGF-1inducedA549cells proliferation.Then the A549cells in the logarithmic growth phase were randomly divided intoone control group （group C） and four treatment groups （group CEL₁、group CEL₂、group CEL₃and group CEL₄）. The cells in group C was cultured commonly for48h.The cells in treatment groups were exposed on6.25μmol/L、12.5μmol/L、25μmol/Land50μmol/L celecoxib, in addition the minimum effective dose of IGF-1respectively for48h. The cell proliferation of each group was detected using MTTassay.（2） Cell invasion assayThe A549cells in the logarithmic growth phase were randomly divided into one control group （group C） and four treatment groups （group CEL₁、group CEL₂、groupCEL₃and group CEL₄）. The cells in treatment groups were cultured with6.25μmol/L、12.5μmol/L、25μmol/L and50μmol/L celecoxib respectively for32h. Thenthe cells were plated in the upper chamber, containing the same concentrations ofcelecoxib. The lower chamber of every groups were filled with40ng/mL IGF-1as achemoattractant. After another16h incubation, the cells invading through the filterwere manually counted.（3） Western Blot analysisThe A549cells in the logarithmic growth phase were randomly divided into onecontrol group （group C） and three treatment groups （group CEL、group I and groupCEL+I）. The cells in group C were cultured commonly for48h. The cells in groupCEL were treated with12.5μmol/L celecoxib for48h. The cells in group I wereexposed on40ng/mL IGF-1for48h. The cells in group CEL+I were cultured with12.5μmol/L celecoxib, in addition40ng/mL IGF-1for48h. The expression ofp-IGF-1R and p-AKT in each group was detected using Western Blot analysis.（4） ELISAanalysisThe A549cells in the logarithmic growth phase were randomly divided into onecontrol group （group C） and four treatment groups （group CEL₁、group CEL₂、groupCEL₃and group CEL₄）. The cells in group C were cultured commonly for48h. Thecells in treatment groups were exposed on6.25μmol/L、12.5μmol/L、25μmol/L and50μmol/L celecoxib respectively for48h. The expression of IGFBP-3ofsupernatants in each group was detected using IGFBP-3Active ELISA KitTM.Results（1）IGF-1stimulation promotedA549cells growth.And the minimum effective doseof IGF-1was40ng/mL （P<0.05）. Compared with group C, celecoxib protectedA549cells against IGF-1induced cell growth （P<0.05）, even the dose of celecoxib was low （6.25μmol/L）.（2）Compared with group C, the numbers of A549cells invading filtered weresignificantly decreased in group CEL₂、group CEL₃and group CEL₄（P<0.05）.And the results showed a dose-dependent manner （P<0.05）.（3）Using40ng/mL IGF-1could promoted phosphorylation of IGF-1R and AKT（P<0.05）.12.5μmol/L celecoxib could inhibited the expression of p-IGF-1Rand p-AKT （P<0.05）. And12.5μmol/L celecoxib could inhibited40ng/mLIGF-1induced phosphorylation of IGF-1R and AKT （P<0.05）.（4）Compared with group C, exposure of A549cells on different concentrations ofcelecoxib could elevated the levels of IGFBP-3in tumor cell culturesupernatants （P<0.05）, even the concentration of celecoxib was very low（6.25μmol/L）. And the results showed a concentration-dependent manner（P<0.05）.Conclusion（1） IGF-1could induce A549cells growth and invasion. The possible mechanism isthat IGF-1promoted phosphorylation of IGF-1R and AKT.（2） Celecoxib protected A549cells against IGF-1induced cell growth and invasion.The mechanism may be that celecoxib inhibit IGF-1induced phosphorylation ofIGF-1R and AKT.（3） Celecoxib inhibited cell growth and invasion of A549cells. The possiblemechanism is that celecoxib inhibit phosphorylation of IGF-1R and AKT. Andcelecoxib up-regulated expression of IGFBP-3. ObjectiveTo study the effects of celecoxib on cell adhesion and expression of CD44v6innon-small cell lung cancer cell line A549.MethodsThe A549cells in the logarithmic growth phase were randomly divided into onecontrol group （group C） and three treatment groups （group CEL1、group CEL2and group CEL3）. The cells in celecoxib groups were exposed to12.5μmol/L、25μmol/L and50μmol/L celecoxib for48h, respectively. Cell adhesion rate wasdetected by adhesion assay, and the expression of CD44v6protein was determinedby Western blot analysis.ResultsCompared with C group， the rates of cell adhesion were significantlydecreased（P<0.05）, the expression of CD44v6protein was down-regulated in aconcentration-dependent manner（P<0.05）in the treatment groups.ConclusionThe celecoxib can inhibit the adhesion of A549cells. This effect of celecoxibmay be related to the down-regulation of CD44v6protein expression.