溶血磷脂酸抑制顺铂诱导的卵巢癌细胞凋亡  

Protective effect of lysophosphatidic acid on apoptosis of human epithelial ovarian cancer cells induced by cisplatin

作　　者：刘红[1] 房朝晖[1] 吴小华 

机构地区：[1]河北医科大学第四医院妇瘤科,石家庄050011 [2]河北医科大学第四医院妇产科,石家庄050011

出　　处：《肿瘤》2008年第7期543-547,共5页Tumor

基　　金：国家自然科学基金资助项目(编号:30572093);河北省自然科学基金资助项目(编号:C2007001059)

摘　　要：目的:探讨溶血磷脂酸(lysophosphatidic acid,LPA)对顺铂诱导的卵巢癌细胞凋亡的抑制作用及其作用机制。方法:体外培养卵巢癌细胞株SKOV3,采用MTT法检测LPA对顺铂(cisplatin,DDP)作用后卵巢癌细胞株SKOV3增殖活性的影响,Hoechst33258荧光染色观察凋亡细胞,用FCM法分析细胞周期变化和细胞凋亡率,DNA片段凝胶电泳观察凋亡细胞的DNA"梯状"条带,免疫细胞化学法及RT-PCR法分别检测细胞凋亡相关蛋白及其mRNA的表达。结果:LPA能降低DDP对SKOV3细胞生长的抑制作用,同时增加G0/G1期细胞比例,降低S期细胞比例和凋亡率。Hoechst33258染色检测示LPA作用后凋亡小体明显减少。DNA片段凝胶电泳示LPA作用后不产生明显的凋亡片段。10μmol/L的LPA作用SKOV3细胞48h后,bcl-2基因及其蛋白表达水平升高,而bax基因及其蛋白表达水平降低(P<0.01)。结论:LPA可通过上调bcl-2基因及蛋白表达,下调bax基因及蛋白表达,抑制DDP诱导的卵巢癌细胞凋亡。因此,针对LPA的治疗有望提高DDP疗效,改善卵巢癌患者的预后。Objective： To investigate the effect of lysophosphatidic acid （LPA） on cisplatin-induced apoptosis in ovarian carcinoma cells and analyze its action mechanism. Methods： Human ovarian carcinoma cell line SKOV3 cells were cultured in vitro, and methyl thiazolyl tetrazolium （MTT） assay was used to examine the effect of cisplatin （DDP） with or without LPA on the proliferation of SKOV3 cells. Hoechst 33258 fluorescence staining was used to observe the apoptotic cells. Flow cytometry （FCM） was used to analyze cell cycle and apoptotic rate. DNA fragment agarose gel electrophoresis was performed to detect the specific DNA ＂ladder＂. The expres- sion of apoptosis associated proteins and their mRNA were determined using immunocytochemistry and RT-PCR. Results： LPA de- creased the growth inhibition induced by DDP. In addition, LPA increased the percentage of cells in G0/G1 phase and reduced the percentage of cells in S phase and the apoptotic rate. Hoechst 33258 staining showed that the number of apoptotic body was significantly decreased. DNA fragment agarose gel extrophoresis demonstrated that no specific DNA ＂ladder＂ was observed after LPA treatment. The expression of bcl-2 mRNA and protein was increased while the expression of bax mRNA and protein was decreased in SKOV3 cells exposed to 10 μmol/L LPA for 48 h （ P 〈 0.01 ）. Conclusion ： LPA could protect SKOV3 cells againt the apoptosis induced by DDP. This protective effect was related with the up-regulation of bcl-2 expression and down-regulation of bax expression at mRNA and protein levels. LPA-targeted therapy may increase the clinical outcome of DDP and improve the prognosis of ovarian cancer patients.

关 键 词：卵巢肿瘤 溶血磷脂酸类 顺铂 细胞凋亡 

分 类 号：R737.31[医药卫生—肿瘤] R730.7[医药卫生—临床医学]