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目的:探讨溶血磷脂酸(LPA)抑制顺铂(DDP)诱导卵巢癌细胞凋亡的机制。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DDP、DDP+LPA和DDP+LPA+PD98059对SKOV3细胞增殖活性的影响;应用Ho-echst33258荧光染色观察凋亡细胞;应用实时荧光定量PCR(RT-PCR)测定单用LPA或合用PD98059对SKOV3细胞环氧化酶-2(COX-2)表达的影响;应用流式细胞仪(FCM)测定细胞凋亡及细胞周期的变化。结果:LPA对DPP抑制卵巢癌细胞增殖有拮抗作用,且增加S期细胞比率;联合应用PD98059后,卵巢癌细胞生长受抑制,S期比率降低,而凋亡小体产生增多。RT-PCR结果显示,LPA能促进COX-2表达(P<0.05),而合用PD98059后COX-2表达降低(P<0.05)。结论:LPA通过MAPK信号传导通路抑制顺铂诱导的卵巢癌细胞凋亡,且与COX-2表达有关。
Objective: To investigate the mechanism of LPA inhibiting cisplatin (DDP) -induced ovarian cancer cell apoptosis. Methods: Human ovarian epithelial carcinoma cell line SKOV3 was cultured in vitro. PD98059, a specific inhibitor of mitogen-activated protein kinase (MAPK) signaling pathway, was selected for determination of DDP, The effect of DDP + LPA and DDP + LPA + PD98059 on the proliferation of SKOV3 cells was observed. The apoptotic cells were observed by Ho-echst33258 staining. The apoptosis of SKOV3 cells was evaluated by real-time fluorescence quantitative PCR (RT-PCR) (COX-2) expression; using flow cytometry (FCM) determination of apoptosis and cell cycle changes. Results: LPA could antagonize the proliferation of ovarian cancer cells induced by DPP and increase the ratio of S phase cells. Combined with PD98059, the growth of ovarian cancer cells was inhibited, while the proportion of S phase was decreased and the apoptotic bodies were increased. 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 with PD98059 (P <0.05). Conclusion: LPA inhibits cisplatin-induced ovarian cancer cell apoptosis through MAPK signaling pathway, and is related to the expression of COX-2.