Antitumor Activity Of C225 In Combination With Docetaxel In Human Gastric Carcinima Cell Lines MGC803 And BGC823 In Vitro

Purpose1,To observe the depressive effect of human gastric carcinoma cell lines MGC803 and BGC823 using the model of Synergy when C225 is uesed only or combined with Docetaxel,which is expected to disuss the Synergy effect as well as an optimal sequence of C225 in combination with Docetaxel .2,To observe the cell cycles distribution and to analysis the expression of EGFR and its downstream proteins,which is expected to disuss the molecular mechanisum of antitumor activity of the optimal sequence for the aim to offer basic information for clinical target therapy of gastric cancer.MethodsMGC803 and BGC823 cells were added in RPMI-1640 medium supplemented with 10%FBS,100u/mL penicillin,and100u/mL streptomycin,cultured in a humidified 5%CO2/95% air atmosphere at 37℃.1,While cells proliferated in logarithm,the effect of C225 alone or combined with Docetaxel on the proliferation of MGC803 and BGC823 cells was evaluated by MTT assay,and Q value was calculated to judge the combined effect. Optimal sequence of C225 in combination with Docetaxel was also been evaluated.2,The cell cycles distribution was test by FCM inC225 and the optimal sequence of C225 in combination with Docetaxel.3,Expression of EGFR and its downstream proteins p-38MAPK and p-AKT were test before and after the administration of optimal sequence of C225 in combination with Docetaxel using Western-blotting.Results1. MTT assay has shown that C225 alone at concentrations up to 1000μg/ml didn't show an obvious concentration-dependent inhabitation on the proliferation of MGC803 and BGC823 cells; Docetaxel alone significantly inhibited proliferation of MGC803 and BGC823 cells in a concentration-dependent manner; C225 enhanced the anti-proliferation effect of Docetaxel on MGC803 and BGC823 cells in a certain degree.The sequencial administration that is Docetaxel were given before C225 is the best sequence of their combination.2. Flow cytometry analysis revealed that C225 induced MGC803 and BGC823 cells G0/G1phase arrest,decresed the S phase cells,and shown an increased poptosis rate.3. Western blotting confirmed C225effectively downregulated p-38MAPK and P-AKT Protein levels so ao to inhibited the proliferation of MGC803 and BGC823 cells.Conclusion1. C225 enhanced the anti-proliferation effect of Docetaxel on human gastric carcinoma cell Lines MGC803 and BGC823 in a certain degree.The sequencial administration that is Docetaxel were given before C225 is the best sequence of their combination.2. The main molecular mechanisum of antitumor activity of the optimal sequence can be explained as follows:Docetaxel up-regulates the expression of EGFR,so as to enhances the inhibitive effect of C225 on EGFR signaling and decreses the expression of downstream proteins p-38MAPK and p-AKT ,which leads to the the increase of anti-proliferation effect of C225 and induced more G0/G1 phase arrest,so that shows more remarkable inhibition on the proliferation of gastric cancer cells.3. The antitumor activity of C225 observed in human gastric cancer cells warrants consideration of its use in clinical treatment regimens for human gastric cancer as a single agent or a combination drug with various chemotherapeutic agents.