Experimental Studies For The Effect Of Multiple Inhibition Of COX-2, 5-LOX And P12-LOX On Human SGC-7901 Cell Line

Objective COX-2,5-LOX and p12-LOX pathways are very important pathways of arachidonic acid metabolism, and there is a close relationship between those pathways and gastric cancer. This article aims to reveal phenomena of dynamic equilibrium in those three pathways and to explore effect of block those pathways on human gastric adenocarcinoma cell SGC-7901 in vitro.Method Enough gastric adenocarcinoma cell SGC-7901 are cultured using general methods. Drug solution of COX-2 selective inhibitor nimesulide,5-LOX selective inhibitor zileuton and p12-LOX selective inhibitor baicalein are prepared using general methods, too. Then, they are randomly divided into seven kinds of experimental groups, namely, nimesulide inhibition group, zileuton inhibition group, baicalein inhibition group, nimesulide/zileuton combined inhibition group, nimesulide/baicalein combined inhibition group, zileuton/baicalein combined inhibition group and three drugs combined inhibition group. Selective inhibitor nimesulide, zileuton, and baicalein are added into the corresponding groups. When the drugs are added, we cultured the cells for another 72 hours.ELISA detection kits are used to detect PGE2, LTB4, and 12S-HETE protein contents in the experimental groups. We compare those changes of protein contents in experimental groups to reveal phenomena of dynamic equilibrium in those three pathways. Inverted microscope is used to observe the changes of sample cells in all seven experiemenal groups. MTT assay kit is used in all experimental groups to detect sample cell survival. We compare the different cell survival in experimental samples to determine the effect of block those pathways on human gastric adenocarcinoma cell SGC-7901 in vitro. In this study, control group is established. The experimental drugs are not added into the samples in the control group. The remaining experimental steps are the same to the experimental groups. Repeat the experimental steps above three times, and mean±SD is calculated.Result In the nimesulide inhibition group, zileuton inhibition group and baicalein inhibition group, with increasing of the experimental drug concentration, protein contents of PGE2, LTB4 and 12S-HETE are respectively decreased (P<0.05). The trends show dose dependent manner. In the nimesulide/zileuton combined inhibition group, nimesulide/baicalein combined inhibition group and zileuton/baicalein combined inhibition group, protein contents of any two metabolic pathways which are added the corresponding drugs are decreased, and protein content of the third pathway is increased (P<0.05).In the nimesulide inhibition group, zileuton inhibition group and baicalein inhibition group, with increasing of the experimental drug concentration, cell suvival levels are respectively decreased (P<0.05). The trends show dose dependent manner. And cell suvival levels are decreased more in the combined inhibition group (P<0.05). The three drugs combined inhibition group shows the lowest survival rate in this study. Inverted microscope is used to observe microscopic changes in cells. In the experimental group, cells are smaller, and are changed into spindle or irregular in shape. The numbers of cells are deceased in different degrees. Edge of the light-emitting cells, round cells, suspension cells and cells which are nearly leave the adherent state are observed. These changes became clear when the drug concentrations are increased. In the control group, adherent cells were polygonal, and little suspension cells are observed.Conclusion Selective inhibitor nimesulide, zileuton and baicalein can inhibit COX-2, 5-LOX and p12-LOX pathways. The trends show dose dependent manner. And three metabolic pathways show dynamic equilibrium and complementary relationship. These results provide a theoretical basis in gastric cancer prevention and treatment.Selective inhibitor nimesulide, zileuton and baicalein were able to significantly reduce cancer cell survival, and the trends show dose dependent manner. Gastric cancer cell survival rate is decreased more significant in the combined inhibition groups than other groups and combined inhibition shows a better anticancer activity. These results provide a real value in gastric cancer prevention and treatment.