The Effect Of Evodiamine On Human Ovarian Cancer Cell Line HO-8910PM

BackgroundOvarian cancer is one of the most common gynecologic cancers, only next to cervical cancer and endometrial cancer. However, the morality rate of ovarian cancer ranks first.5year survival rate is poor despite routine surgery and chemotherapy. Therefore, it is critical to develop effective measures against ovarian cancer. Evodiamine is a kind of quinolone alkaloid isolated from a traditional Chinese medicinal plant known as Wu-Chu-Yu. Evodiamine possesses pharmacological effects of anti-inflammatory, antinociceptive and vasorelaxant properties. Recent studies have shown that evodiamine has inhibitory effects on various tumors including human prostate cancer cell LNCaP, PC-3, human melanoma cell A375-S2, human leukemic cell U937and human cervical cancer cell HeLa. However, the effect of evodiamine on ovarian cancer cell is still unclear.ObjectiveTo explore the effect and the mechanism of evodiamine on proliferation, cell cycle, apoptosis, migration and invasion of ovarian cancer cell HO-8910PM. Methods1.3-(4,5-dimethiylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect the growth inhibition rate of evodiamine in HO-8910PM cells.2. Cell cycle was determined by FCM.3. Apoptosis induction was assessed by Annexin V-FITC/PI double staining assay.4. Transwell tramber assay was used to examine the effect of evodiamine on cell migration and invasion on HO-8910PM cells.5. Western blot analysis was performed to detect cell cycle, apoptosis, migration and invasion related protein.Results1. Evodiamine can inhibit the proliferation of ovarian cancer cell. After incubation for24h, the growth inhibition was not obvious compared with the control group (p>0.05). The significant inhibitory effect on HO-8910PM cell growth caused by evodiamine was observed at48h and persisted for96h in time-dependent and concentration-dependent way(p<0.05).2. Evodiamine can induce cell cycle arrest of ovarian cancer cell. We assessed cell cycle distribution by PI staining after treatment with evodiamine (0～5μM) for24h. The number of cells during the various phases was significantly different (P＜0.05) between the control and experimental groups. With the concentration of evodiamine increasing, G2/M percentage was dramatically increased while G0/G1percentage was significantly decreased (P<0.05). To explore the mechanism of G2/M arrest caused by evodiamine, western blot was performed to measure the expressions of cell cycle-regulating molecules. Compared with the control group, the expression of cyclinB1was significantly increased.3. Evodiamine can induce apoptosis in ovarian cancer cells. HO-8910PM cells treated with evodiamine for48h displayed much higher apoptosis rate than the control group in a does-dependent manner (p<0.05). The underlying mechanism was further explored. After treatment with evodiamine for24h, the activation of caspase3,8,9and PARP was detected as well as the decrease of Bcl-2and the increase of Bax.4. Evodiamine inhibited the migration and invasion of ovarian cancer cells. In the Transwell migration assay, the cells migrated to the lower chambers was reduced by evodiamine in a concentration-dependent manner. In the Transwell invasion assay, the number of cells invaded through the Matrigel-coated filter was reduced by evodiamine dose-dependently. The underlying mechanism was further explored. After treatment with evodiamine for24h, evodiamine down-regulated the expression and activity of FAK and MMP-2but had no effect on the expression of MMP-9.5. Evodiamine treatment could attenuate the PI3K/Akt signaling and MAPKs signaling. The expression level of PI3K, p-PI3K, Akt, p-Akt, ERK1/2, p-ERK1/2, p38MAPK in HO-8910PM cells was down-regulated after treatment with evodiamine.ConclusionsEvodiamine can inhibit the growth of ovarian cancer cell through G2/M arrest and intrinsic and extrinsic apoptosis. In addition, evodiamine can inhibit its migration and invasion. Furthermore, PI3K/Akt, ERK1/2MAPK and p38MAPK signaling may be involved in the effect of evodiamine.