溶血磷脂酸促卵巢癌细胞增殖机制的研究

Mechanisms involved in lysophosphatidic acid inducing proliferation of ovarian carcinoma cells

目的探讨溶血磷脂酸(LPA)促进卵巢癌细胞增殖的机制。方法体外培养人卵巢上皮癌细胞株SKOV3,选用促分裂素原活化蛋白激酶(MAPK)信号传导通路特异性阻断剂PD98059,应用四甲基偶氮唑蓝(MTT)比色法测定LPA和PD98059对SKOV3细胞增殖活性的影响;应用实时荧光定量PCR(RT-PCR)测定LPA单独或合用PD98059对SKOV3细胞表达环氧化酶-2(COX-2)的影响;应用流式细胞仪(FCM)测定细胞的凋亡及细胞周期的变化。结果 LPA(浓度>10μmol/L)以剂量依赖方式促进SKOV3细胞增殖,PD98059抑制LPA促增殖作用(P<0.05)。RT-PCR结果显示,LPA能促进COX-2的表达(P<0.05),而合用PD98059后COX-2的表达降低(P<0.05)。FCM结果显示,LPA能促进细胞增殖,抑制细胞凋亡,增加S期比率,而合用PD98059后LPA促增殖作用消失,且G0/G1期比率升高,S期比率降低。结论 LPA通过MAPK信号传导通路促进卵巢癌细胞增殖,且与COX-2表达有关。

Objective To investigate the mechanism of lysophosphatidic acid（LPA） inducing proliferation of ovarian carcinoma cells.Methods The human ovarian cancer cell line SKOV3 was cultured in vitro,and PD98059 which could inhibit the mitogen-activated protein kinase（MAPK） signaling pathway was used.Proliferative activity of SKOV3 cells stimulated by LPA with or without PD98059 was assessed by Methyl thiazolyl tetrazolium（MTT）.Expression of COX-2 in SKOV3 cells was assessed by RT-PCR.Apoptosis and cell cycle of SKOV3 cells were detected by flow cytometry（FCM）.Results LPA（when density 10 μmol/L） could promote proliferation of SKOV3 cells in a dose-dependent manner,while PD98059 could inhibit the effect（P〈0.05）.RT-PCR showed that LPA could increase expression of COX-2 in SKOV3 cells（P〈0.05）,while PD98059 could decrease the effect（P〈0.05）.FCM showed that LPA could promote proliferation,inhibit apoptosis,and increase the proportion of cells in the S phase.However,combined with PD98059 in culture,cell proliferation and proportion of cells in the S phase were significantly decreased,while the proportion of cells in the G0/G1 phase was increased.Conclusion LPA may promote proliferation of ovarian carcinoma cells via the MAPK signaling pathway,and the mechanism is related to expression of COX-2.