The Effect Of Epoxide Hydrolase Enzyme Inhibitor Ns-398 Combined With Oxaliplatin On Proliferation And Apoptosis Of Hela Cells

Objective: Among genital malignant tumor, cervical cancer with the highest incidence rate greatly affects women’s health. In recent years, the incidence of cervical cancer rises every year and age of onset was younger trend. It is clinically urgent to solve this disease. Now, chemotherapy combined with operation is the main method for the treatment of cervical cancer, and traditional chemotherapeutics with many toxic or side effects effects the life quality of patients. Searching for an effective chemotherapeutics with little side-effect is the hot research topic in clinic. In recent years, the researches indicated that NS-398, inhibitor of COX-2, could inhibit the activity of COX-2 to inhibit the growth of cancer cells. The study mainly research on the effect of NS-398, oxaliplatin and both on proliferation and apoptosis of Hela cells to provide theoretical basis for cervical cancer treatment.Method: CCK-8 method was used to detect the inhibition rate of L-OHP, NS-398 at different concentration and both. Observed the morphologic of cells treated with L-OHP( 10mg/L), NS-398( 60 μmol/L) at different concentration and both. Flow Cytometry was used for the detecting apoptosis rate and cell cycle. Rhodomine 123 staining kit and Double immunofluorescence staining is exploited to detect the apoptosis of Hela cells treated with L-OHP and NS-398. Apoptosis was measured using DNA fragmentation. Caspase-3 activity of Hela cells treated with L-OHP, NS-398 and both was detected using caspase-3 activity assays.Results:1 Results of CCK-8 indicated that L-OHP and NS-398 could inhibit the Hela cells(P<0.05), and the inhibition effect has dose dependency. Jointaction is effective than single drug.2 The observed results of inverted microscope indicated that Hela cells treated with L-OHP(10mg/L), NS-398(60 μmol/L)and both shrinked and turned round, cells fall off bottom of the cell culture bottle, cell ruptured and contents overflowed.3 Rhodomine 123 staining results indicated that the fluorescent intensity of Hela cells treated with L-OHP(10mg/L), NS-398(60 μmol/L)and both for 24 h was stronger, the cells treated with both is the most obvious.4 Results of flow cytometry showed that the apoptosis rate Hela cells treated with L-OHP(10mg/L), NS-398(60 μmol/L)and both for 24 h separately was:(10.3±0.06)%,(10.5±0.14)%,(15.7±0.13)%, cells treated with drugs had significant differences(P<0.01) as compared with that of negative control group(2.4±0.36)%. Results also showed that L-OHP and NS-398 could markedly arrest cell cycle in G1 phase.5 Agarose gel electrophoresis results showed that: after cells treated with L-OHP(10mg/L)and NS-398(60 μmol/L)for 48 h, Ladder’s band was seen.6 Testing results show that after cells treated with L-OHP(10mg/L), NS-398(60 μmol/L)and both for 48 h, caspase-3 was activated. Caspase-3 activity of cells in L-OHP(10ml/L) group increased to 1.5 times of that in negative control group. Caspase-3 activity of cells in NS-398(60 μmol/L) group increased to 1.43 times of that in negative control group. The caspase-3 activity of cells treated with both increased to 2.48 times of that in negative control group, all has distinct difference between normal cells and cells treated with drug(P<0.01).Conclusion:1 L-OHP and NS-398 both could inhibit Hela cells, the inhibition effect has dose dependency. Joint action is effective than single drug.2 The inhibition effect of L-OHP and NS-398 is relevant to their function of inducing apoptosis.3 L-OHP and NS-398 could affect the cell cycle of Hela cells, Cell cycle was blocked in the G1 phases.4 L-OHP and NS-398 could induce the apoptosis of Hela cells, which is relevant to the activization of caspase-3.