丹皮酚和阿司匹林对人结肠癌LoVo细胞增殖的影响及其机制

Effect of Paeonol on in Proliferation of Human Colorectal Cancer LoVo Cells and Its Mechanism

目的:探讨丹皮酚和阿司匹林和阿司匹林对人结肠癌细胞系LoVo细胞增殖的影响及可能作用机制。方法:用不同浓度丹皮酚(15.63-250mg/L)和阿司匹林(98.08-1 801.6mg/L)处理体外培养的LoVo细胞24,48,72,96h,CKK-8比色法测定其对结肠癌LoVo细胞的活力的影响;根据CKK-8结果分为丹皮酚组(浓度31.25,62.50,125mg/L)和阿司匹林组(浓度分别为180.16,900.80,1 801.6mg/L),进一步处理48h后行流式细胞仪检测细胞凋亡、激光共聚焦显微镜检测细胞内Ca2+浓度的变化及RT-PCR法检测RUNX3基因表达情况。结果:丹皮酚(浓度15.63-250mg/L)和阿司匹林(浓度90.08-1 801.6mg/L)均能能显著降低LoVo细胞存活率,呈剂量、时间依赖性;丹皮酚组处理细胞48h后可诱导细胞凋亡,流式细胞仪检测凋亡率分别为13.5%,21.4%,34.6%,明显高于对照组;阿司匹林组细胞凋亡率分别为14.8%,25.8%,37.7%,明显高于对照组;丹皮酚组荧光强度分别为41.36±4.62,57.51±3.83和69.43±3.76;阿司匹林组荧光强度分别为38.24±4.62,53.31±4.92和65.64±5.25,丹皮酚组、阿司匹林组与对照组(荧光强度24.45±3.74)比较,差异均有统计学意义(P<0.05);丹皮酚和阿司匹林均能上调RUNX3mRNA的表达,呈剂量依赖性。结论:丹皮酚、阿司匹林均能抑制LoVo细胞的增殖并诱导其凋亡,其作用机制可能与增加细胞内Ca2+含量和上调RUNX3基因的表达有关。丹皮酚与阿司匹林可能具有相似抗大肠癌细胞作用机制。

Objective：To investigate the effect of paeonol and aspirin on the proliferation of colorectal cancer line LoVo and discuss their possible mechanisms.Methods：LoVo cells were cultured in vitro.After treatment by paeonol and aspirin at different concentrations respectively at the end of24,48,72 and 96h,the cell survival was determined by the CKK-8method.Apoptosis was detected by flow cytometry.The concentration of intracellular calcium was investigated by laser confocal scanning microscope with calcium-fluorescent probes-Fluo-3/AM.Reverse transcriptionpolymerase chain reaction（RT-PCR）was used for mRNA analysis.Results：From the data of CKK-8,the cell proliferation of human colorectal cancer LoVo cells was inhibited by paeonol and aspirin in a dose-dependent and time-dependent manner.Flow cytometry assays showed that pae-onol and aspirin both significantly induced apoptosis in LoVo cells.After treated with paeonol for48 h,the apoptosis rate of Lovo cells was 13.5%,21.4% and 34.6% respectively,and after treated with aspirin,the apoptosis rate of Lovo cells was 14.8%,25.8%and 37.7%respectively,which showed an obvious concentration-effect relationship.The Ca2＋relative fluorescence intensity of paeonol group was 41.36±4.62,57.51±3.83,and 69.43±3.76 respectively,and the Ca2＋relative fluorescence intensity of aspirin group was 38.24±4.62,53.31±4.92,and 65.64±5.25 respectively.The concentration of intracellular calcium was significantly elevated by paeonol and aspirin.RT-PCR showed that paeonol and aspirin up-regulated RUNX3 in a dose-dependent manner in LoVo cells.Conclusion：Paeonol as well as aspirin can inhibit the proliferation of LoVo cells and induce apoptosis,and the mechanism of paeonol on apoptosis may be related to the upregulation of RUNX3 expression and the increase of concentration of intracellular calcium.The mechanism of paeonol and aspirin may be similar.