Effects Of BFGF And Celecoxib On Proliferation And Apoptosis Of Gastric Carcinoma BGC-823 Cells

Gastric cancer is secondary common malignant tumor of the world.It is important to study the development of gastric cancer. It has all-important sense to prevent and cure gastric cancer and precancerosis.bFGF has close relation with tumorigenesis.NSAIDs is another hot study in treatment and prerention of tumour for the past few years.It can cut down chanciness in gastroenteric tumor.celecoxib is a kind of NSAIDs.How to effect on Apoptosis of gastric cancer cells by celecoxib is still not clear now.We took exogenous bFGF and celecoxib on BGC-823 cells.To observe the changes of grouth,activities of PKB, expressions of COX-2,NF-κB and VEGF protein in BGC-823 cells and the effects of PI3K/PKB signal transduction in the process.To approach the effects of bFGF and celecoxib take in pathogenesy of gastric cancer and the interaction of bFGF and celecoxib.Methods1 Apoptosis of BGC-823 gastric cancer cells induced by celecoxib were observed by fluorescence microscope.2 Effects of bFGF,wortmannin(the inhibitor of PI3K )and celecoxib on proliferation of BGC-823 gastric cancer cells were detected by MTT assay.3 Apoptosis of BGC-823 gastric cancer cells induced by celecoxib were detected by FACScan flow cytometer.4 Changes of PKB activity induced by bFGF and expressions of COX-2,NF-κB and VEGF protein induced by bFGF and celecoxib were detected by western blot.Results1 Effects of PKB activity induced by bFGF:different density bFGF up-regulated PKB activity in gastric cancer BGC-823 cells.25ng/ml bFGF induced the maximal expressions of p-PKB.Treated with 25ng/ml bFGF for 5,10,30, 60min respectively,activity of PKB in BGC-823 cells began to increase at 5min and reached the peak at 10min.The difference between bFGF groups and control group have statistics significance (P<0．01) . Pre-treated with wortmannin,PKB activity decreased compared with control group,which has statistics significance (P<0．01) .2 MTT assay showed that:treated BGC-823 cells with bFGF,the cell proliferative rate increased (P<0．05) .Treated BGC-823 cells with celecoxib,the cell inhibition ratio increased dose dependently(P<0．05) .3 After being treated with different density of celecoxib,the shapes of BGC-823 cell had changed markedly.Parts of attached cells shrinked and changed round.At last they floated.Observed by fluorescence microscope showed that after being treated with celecoxib the shapes of BGC-823 cells changed irregularly and apoptosis appeared with time pasted and density of celecoxib increased.4 Flow cytometry showed that celecoxib induced apoptosis of BGC-823 obviously at 24h.It had positive correlation with density of celecoxib (r=0．99,P<0．05) . Apotosis of BGC-823 cells in control group,25μmol/l group and 50μmol/l group were 2．39%,33．69%and 59．98%.5 Western blot showed that:(1) bFGF time dependently induced expressions of COX-2,NF-κB and VEGF protein.The peak of expressions of COX-2,NF-κB and VEGF protein were at 24h ,12h and 24h respectively.The difference between each action time treated with bFGF and control group have statistics significance(P<0．01) .(2) After being treated with celecoxib, expressions of COX-2,NF-κB and VEGF protein were decreased in dose dependently.The difference of expressions of COX-2,NF-κB and VEGF protein between control group, 25μmol/l group and 50μmol/l group has statistics significance(P<0．01) .There are associativity among COX-2,NF-κB and VEGF (r=0．852, 0．908 and 0．930 respectively P < 0．01) .(3) Celecoxib had depressant effect in expressions of COX-2,NF-κB and VEGF in BGC-823 cell pre-treated with bFGF (P＜.05) . 1 bFGF promoted proliferation,resisted apoptosis induced by starvation in gastriccancer BGC-823 cells through PI3K/PKB pathway.2 bFGF up-regulated PKB activity which was inhibited by wortmannin.3 bFGF induced expressions of COX-2,NF-κB and VEGF protein in BGC-823 gastric cancer cells through PI3K/PKB pathway.4 Celecoxib induced apoptosis and decreased expressions of COX-2,NF-κB and VEGF protein in gastric cance BGC-823 r cells.5 Celecoxib had depressant effect on proliferation of BGC-823 cells induced by bFGF.