阿司匹林对肝癌细胞生长的抑制作用及其机制

目的 观察阿司匹林(AS)对肝癌细胞SMMC-7721生长的作用,并探讨其作用机制.方法 采用噻唑蓝(MTT)比色法检测不同浓度阿司匹林对肝癌细胞细胞生长的抑制作用以及流式细胞仪检测不同浓度阿司匹林对肝癌细胞细胞周期的影响;免疫组织化学方法检测阿司匹林对肝癌细胞细胞增殖核抗原(PCNA)、环氧合酶-2(COX-2)表达的影响;并用逆转录-聚合酶链反应(RT-PCR)测定COX-2基因的表达.结果 肝癌细胞经不同浓度的阿司匹林作用后,其生长抑制率明显升高(P<0.05),且与阿司匹林浓度有关和作用时间相关.细胞周期检测结果显示,经不同浓度阿司匹林作用后的SMMC-7721细胞,细胞周期发生明显变改.表现为随着阿司匹林浓度的增加,S期细胞比例下降明显,G0/G1期及G2/M期细胞比例则上升(10-2、10-3moL/L AS组与对照组比较P<0.05).免疫细胞化学染色显示,阿司匹林能下调肝癌细胞COX-2、PCNA的表达.RT-PCR检测表明,COX-2基因表达电泳条带与对照组比较辉度明显降低,其中10 mmol/L阿司匹林组最低,说明随着阿司匹林浓度增加,COX-2表达逐渐减少,阿司匹林能下调SMMC-7721细胞表达COX-2基因.结论 阿司匹林能有效抑制肝癌细胞SMMC-7721的生长,这种作用可能是其通过抑制细胞增殖,与抑制COX-2、PCNA的表达有关.

PCNA inhibition effect of aspirin on the growth of hepatoceUular carcinoma MMC-7721 and the mechanism therein involved

Objective To investigate the effect of aspirin on the growth of Hepatocellular Carci-noma(HCC) in vitro and the relevant mechanism. Methods SMMC-7721 HCC cells were randomly di-vided into four groups, after exposured to aspirin with different concentration ranged from 1 × 10-2 to 1 ×10-4 mol/L,The suppressive effect of aspirin on MMC-7721 HCC ceils was measured by MTI" method;the cell cycle was analyzed by flow cytometry. PCNA and COX-2 protein was examined in MMC-7721 HCC cells by immunohistochemistry. The expression of COX-2 gene was assayed by RT-PCR. Results After exposured to aspirin,the suppression ratio of SMCC-7721 cells was increased (P < 0.05). Aspirin could inhibit DNA syntheses. FCM data showed that,among phases of cell cycle,the G0/G1 and G2/M phase ra-tio were increased and S phase ratio was inhibited significantly(P < 0.05). Immunohistochemistry indica-ted that aspirin effectively decreased COX-2 and PCNA expression in MMC-7721 HCC cell. Aspirin could inhibit the expression of COX-2 gene. Conclusion Aspirin is effective for inhibiting the growth of HCC.The anti-neoplastic effect aspirin produces may involve the inhibition of COX-2 and PCNA expression.