阿司匹林对卵巢癌SKOV3细胞凋亡的影响及其作用机制研究

Inhibitory effect of aspirin on ovarian carcinoma cells and related molecular mechanism

目的:探讨阿司匹林对卵巢癌细胞凋亡的影响以及对凋亡相关基因bcl-2/bax mRNA的作用。方法:用四甲基偶氮唑蓝(MTT)快速比色法分析阿司匹林对卵巢癌细胞株SKOV3生长的抑制作用,用流式细胞术分析方法定量检测细胞凋亡及细胞周期的变化。用RT-PCR法检测凋亡相关基因bcl-2/bax mRNA的表达。结果:阿司匹林对SK-OV3细胞的增殖有明显的抑制作用,并呈量效和时效的关系;1、5、10mmol/L3个浓度的阿司匹林处理细胞48h后,细胞凋亡率呈剂量依赖性增加,而且G0/G1期细胞比例下降,G2/M期细胞比例升高,细胞被阻滞在G2/M期,细胞周期也发生明显改变;阿司匹林可抑制卵巢癌细胞的bcl-2mRNA表达,而上调bax mRNA的表达。结论:阿司匹林能抑制卵巢癌SKOV3细胞增殖,诱导其凋亡。此作用可能与阿司匹林抑制细胞bcl-2mRNA的表达和上调bax mRNA表达有关。

Objective：To investigate inhibitory effect of aspirin on proliferation of SK- OV3 cells and cellular apoptosis and to detect the expression of bcl-2 and bax gene in SKOV3 cells treated by aspirin in order to investigate related molecular mechanisms, Methods：Proliferation was determined spectophotometrically by the incorporation of tetrazolium dye;optical density at 490nm was then determined using an enzyme-linked immunosorbent reader. Cell apoptosis and cell cycle was detected by cell flow cytometry （FCM）. Bcl-2 and bax mRNA expressions were detected by reverse transcription polymerase chain reaction （RT-PCR）. Data were presented as x ± s and analyzed with SPSS statistical program. A level of P 〈 0.05 was considered statistically significant. Results： （ 1 ） MTr assay showed incubation with any concentration of aspirin for 24hours resulted in no inhibition in SKOV3 cells （P 〉 0.05 ）;but for 48h,72h, 96h, aspirin inhibited SKOV3 tumor cell growth in dose-dependent fashion and in time-dependent fashion;（2）Aspirin induced cellular apoptosis ：AP（ apoptosis percentage） detected by FCM （ flow cytometry）, showed that aspirin with 0,1,5,10mmol/L for 48hours induced cell apoptosis in a dose-dependent fashion ranging from 3.58% ±0.74%, 11. 13%± 0.81%, 17.90% ± 0. 43% ,30.83% ± 0.44% （ P 〈 0.01 ）. There were visible apoptotic peaks of hypodiploid on the DNA bargraph ; （ 3 ） Compared with control group, after being treated with aspirin, SKOV3 cells of G0/G1 stage was decreased,and cells of G2/M stage was increased; （4）Aspirin can down-regulate the expression of bcl-2 mRNA, up-regulate the expression of bax mRNA. Conelusions：This study showed that aspirin may inhibit the proliferation, and induct apoptosis of SKOV3 cells. Induction of apoptosis was one of mechanism of treatment of ovarian cancer SKOV3 with aspirin. This study first showed that related molecular mechanism of treatment of ovarian cancer using aspirin were down-regulation of bcl-2 mRNA and up-regulation of bax mRNA.