Effects Of Paeonol On Growth,Migration,And Invasion Of Human Gastric Cancer BGC823 Cells

Objective:This study aims to research effects of Paeonol on the growth, migration and invasion of human gastric cancer BGC823 cells, and its relative mechanism.Methods:Human gastric cancer BGC823 cells were cultured normally at first, and when the cells reached logarithmic growth phase, different concentrations of Paeonol was used to intervene the culture for experiment.1. After human gastric cancer BGC823 cells were cultured 24h and 48h respectively with medium contained different concentrations of Paeonol (0,0.05,0.1, 0.2,0.4,0.6,0.8mg/ml), CCK-8 assay was used to detect proliferations of the cells.2. Human gastric cancer BGC823 cells were divided into four groups:control group without Paeonol, Paeonol low-concentration group, Paeonol middle-concentration group and Paeonol high-concentration group. According to the results of CCK-8 assay, low, middle and high concentrations of Paeonol was determined as 0.1,0.2, and 0.4 mg/ml respectively.3. Cells in the four groups were cultured 24h with medium contained cor-responding concentrations of Paeonol (0,0.1,0.2,0.4 mg/ml). Then, flow cyt-ometry (FCM) with Annenin V-FITC/PI double staining method was used to d etect apoptosis of BGC823 cells in each group.4. After BGC823 cells in the four groups were cultured 24h and 48h respectively with medium contained corresponding concentrations of Paeonol, cell migration of BGC823 cells in each group were determined by cell wound healing method.5. After BGC823 cells in the four groups were cultured 24h with medium contained corresponding concentrations of Paeonol, invasion of BGC823 cells in each group were determined by transwell chamber assay.6. After BGC823 cells in the four groups were cultured 48h with medium contained corresponding concentrations of Paeonol, MMP-2 and MMP-9 expressions of BGC823 cells in each group were determined by Western blot technology.Results:1. CCK-8 assay results showed that 24h and 48h inhibitory rates of Paeonol on BGC823 cells were increased gradually with the rise of Paeonol concentration. When concentration of Paeonol was 0.4mg/ml,24h and 48h inhibitory rates of Paeonol on BGC823 cells were 53.3±2.1%,56.1±2.4% respectively. However, when concentration of Paeonol reached 0.6mg/ml,24h and 48h inhibitory rates of Paeonol on BGC823 cells were 68.3±1.6% and 80.1±6.0%, showing obvious cytotoxicity. According to the results of CCK-8 assay, low-concentration, middle-concentration and high-concentration of Paeonol were determined as 0.1 mg/ml,0.2 mg/ml,0.4 mg/ml respectively.2. Observed by microscope, BGC823 cells in the control group showed clear shape and arranged orderly. But compared with BGC823 cells in the control group, those in the three Paeonol groups showed morphological change, and especially those in Paeonol high-concentration group which changed most obviously with more floating cells, unclear shape and disorderly arrangement.3. FCM results showed that cell apoptosis rates of the three Paeonol groups increased obviously compared with that of the control group, and cell apoptosis rate of Paeonol high-concentration increased more than those of Paeonol low-concentration and Paeonol middle-concentration (P< 0.05 or P< 0.01).4. Cell wound healing method results showed that the 24h and 48h healing rates of the three Paeonol groups decreased obviously compared with those of the control group, the 24h and 48h healing rates of Paeonol high-concentration and Paeonol middle-concentration decreased more than those of Paeonol low-concentration (P< 0.05 or P< 0.01).5. Transwell chamber assay showed that the cell numbers crossing chamber in the three Paeonol groups decreased obviously compared with that of the control group (P<0.01), and the cell numbers crossing chamber in the Paeonol high-concentration and Paeonol middle-concentration decreased more than that of Paeonol low-concentration (P<0.05), and the cell numbers crossing chamber in the Paeonol high-concentration decreased more than that of Paeonol middle-concentration (P< 0.05).6. Western blot results showed that MMP-2 and MMP-9 expressions of BGC823 cells in the three Paeonol groups decreased obviously compared with that of the control group, compared with those of Paeonol high-concentration and Paeonol middle-concentration decreasing more than that of Paeonol low-concentration (P<0.05 or P<0.01), and that of Paeonol high-concentration decreasing more than that of Paeonol middle-concentration (P< 0.05).Conclusion:Paeonol can inhibit the growth, migration and invasion of human gastric cancer BGC823 cells with obvious dose-effect relationship, and downregulating expressions of MMP-2 and MMP-9 could be its possible mechanism.