Mechanisms Involved In Lysophosphatidic Acid Induced Occurrence Of Ovarian Carcinoma Cells

ObjectiveThis study aimed to investigate the effect of Lysophosphatidic acid (LPA) on the proliferation of ovarian carcinoma cells, and PD98059which could inhibit mitogen-activated protein kinases (MAPK) signaling pathway was used, when the MAPK signaling pathway was inhabited, we saw the changes of the effect of LPA, and then investigate the mechanisms.MethodsHuman ovarian cancer cell line (SKOV3) was cultured in vitro, and PD98059which could inhibit MAPK signaling pathway was used. The cultured SKOV3cells were divided into4groups randomly. Normal control group (NC group, injected with normal culture medium), Lysophosphatidic acid group (LPA group, injected with LPA under different concentrations), PD98059group (PD98059group, injected with PD98059under the concentration of10μmol/L) and Lysophosphatidic acid+PD98059group (LPA+PD98059group, injected with LPA under different concentrations and PD98059under the concentration of10μmol/L). The proliferative activity of SKOV3cells stimulated by LPA with or without PD98059was assessed by Methyl thiazolyl tetrazolium (MTT). The expression of COX-2in SKOV3cells was assessed by RT-PCR. The apoptosis and cell cycle of SKOV3cells were detected by Flow cytometry (FCM).ResultsLPA (when density>10μmol/L) could promote proliferation of SKOV3cells in a dose-dependent manner, while PD98059could inhibit the effect (P<0.05). RT-PCR showed that LPA could increase the expression of COX-2in SKOV3cells (P<0.05) while PD98059could decrease the effect (P<0.05). FCM showed that LPA could promote proliferation and inhibit apoptosis, and increase the proportion of cells in the S phase of the cell cycle. However, combined with PD98059in culture, the proliferation was significantly decreased as well as cells in the S phase, while the proportion of cells in the Go/G1phase of the cell cycle was increased.ConclusionLPA could promote proliferation and inhabit apoptosis of SKOV3cells in a dose-dependent manner. When PD98059which could inhibit MAPK signaling pathway was used, the effect of LPA was disappeared and the expression of COX-2was decreased. LPA may promote proliferation in ovarian carcinoma cells via MAPK signaling pathway, which was related to the expression of COX-2. ObjectiveThis study aimed to investigate the effect of Lysophosphatidic acid (LPA) on the apoptosis of ovarian carcinoma cells induced by cisplatin, and PD98059which could inhibit mitogen-activated protein kinases (MAPK) signaling pathway was used, when the MAPK signaling pathway was inhabited, we saw the changes of the effect of LPA and cisplatin, and then investigate the mechanisms of Lysophosphatidic acid inhibited apoptosis induced by cisplatin in ovarian carcinoma cells.MethodsHuman ovarian cancer cell line (SKOV3) was cultured, and MAPK inhibitor (PD98059) was used. The cultured SKOV3cells were divided into7groups randomly. Normal control group (NC group, injected with normal culture medium), Cisplatin group (DDP group, injected with cisplatin under different concentrations), Lysophosphatidic acid group (LPA group, injected with LPA under the concentration of40μmol/L), PD98059group (PD98059group, injected with PD98059under the concentration of10μmol/L), Cisplatin+Lysophosphatidic acid group (DDP+LPA group, injected with cisplatin under different concentrations and LPA under the concentration of40μmol/L), Cisplatin+PD98059group (DDP+PD98059group, injected with cisplatin under different concentrations and PD98059under the concentration of10μmol/L)and Cisplatin+Lysophosphatidic acid+PD98059group (DDP+LPA+PD98059group, injected with cisplatin under different concentrations and LPA under the concentration of40μmol/L and PD98059under the concentration of10μmol/L).The proliferation of SKOV3cells stimulated by DDP, DDP+LPA and DDP+LPA+PD98059was assessed by Methyl thiazolyl tetrazolium (MTT). Cell apoptosis was detected by Hoechst33258. The expression of COX-2in SKOV3cells was assessed by RT-PCR. The apoptosis and cell cycle of SKOV3cells were detected by Flow cytometry (FCM).ResultsLPA could antagonize the inhibitory effect of DDP on the proliferation of SKOV3cells. When PD98059which could inhibit MAPK signaling pathway was used, the effect of LPA was disappeared. RT-PCR showed that LPA could increase the expression of COX-2in SKOV3cells (P<0.05), while used with PD98059the expression of COX-2decrease (P<0.05). FCM showed that DDP could induce apoptosis of SKOV3cells and increase the proportion of cells in the S phase of the cell cycle. However, LPA could significantly weaken these effects of DDP and when MAPK signaling pathway was inhibited, the effect of LPA disappeared. Fluorescence microscope showed that obvious apoptotic changes were present in the treatment of SKOV3cells with DDP, and cell nucleus were shrunk, part of the nuclear was fragmented, dense and strong fluorescence and typical apoptotic bodies were visible. While used with lysophosphatidic acid, the effect of DDP was disappeared, apoptotic cells were reduced, the fluorescence was weaker and normal cells were increased. When MAPK signaling pathway was inhibited, the effect of LPA disappeared.ConclusionLPA could inhibit the apoptosis induced by DDP. However, combined with PD98059which could inhibit MAPK signaling pathway in culture, the effect of LPA was disappeared and the expression of COX-2in SKOV3cells was decrease. LPA may inhibit the apoptosis induced by DDP through MAPK signaling pathway, which was related to the expression of COX-2.