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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) on proliferation of ovarian cancer cells. Methods SKOV3 human ovarian epithelial carcinoma cell line was cultured in vitro. PD98059, a specific inhibitor of mitogen-activated protein kinase (MAPK) signaling pathway, was used to detect the expression of LPA and PD98059 by MTT assay On the proliferation of SKOV3 cells. The effect of LPA alone or in combination with PD98059 on the expression of cyclooxygenase-2 (COX-2) in SKOV3 cells was detected by real-time fluorescence quantitative PCR (RT-PCR) Cell apoptosis and cell cycle changes were measured. Results LPA (> 10μmol / L) promoted the proliferation of SKOV3 cells in a dose - dependent manner. PD98059 inhibited the proliferation of SKOV3 cells (P <0.05). The results of RT-PCR showed that LPA could promote the expression of COX-2 (P <0.05), while the expression of COX-2 decreased after treated with PD98059 (P <0.05). FCM showed that LPA could promote cell proliferation, inhibit apoptosis and increase the ratio of S phase. However, PD98059 combined with PDA could not promote the proliferation of LPA, and the ratio of G0 / G1 phase increased and the ratio of S phase decreased. Conclusion LPA can promote the proliferation of ovarian cancer cells through the MAPK signal transduction pathway and is related to the expression of COX-2.