辛基酚影响MCF-7细胞凋亡及bcl_2、caspase3表达

Effects of octylphenol on apoptosis of MCF-7 and expression of Bcl_2 and caspase 3

目的观察辛基酚(OP)对乳腺癌MCF-7乳腺癌细胞凋亡的影响。方法以流式细胞分析、DNA断端末端标记(TUNEL)试验、DNA片断率分析、流式细胞仪免疫荧光和荧光定量PCR等方法观察OP对MCF-7细胞凋亡、bcl2、bax、caspase3表达的影响。结果8、16μmol/LOP作用MCF-7细胞,TUNEL细胞阳性率和DNA片断百分率增加,48h时的凋亡率分别为(6.6±1.32),(16.3±3.48)[对照(4.3±1.18)];bcl2荧光量分别为(63.5±5.84),(30.3±2.49)[对照(57.2±4.07)]。4μmol/LOP明显上调MCF-7细胞中bcl2 mRNA表达;16μmol/LOP则明显降低细胞中bcl2 mRNA表达,且bax、caspase3 mRNA表达明显增加。结论8、16μmol/LOP能明显诱导MCF-7乳腺癌细胞的凋亡,增加DNA片断百分率,其机制可能是通过下调细胞中bcl2的表达、上调bax、caspase3的表达来实现。

[Objective] To observe the effect of octylphenol （OP） on apoptosis of MCF-7 breast cancer cells, [Methods] Flow cytometry, TUNEL（terminal dUTP nick-end labeling） assay, DNA fragment percent analysis, immunofluoreseence by flow cytometry and fluorescent quantitative PCR were used to measure the changes of apoptosis of MCF-7 breast cancer cells and expressions of Bcl2, Bax and caspase 3 when MCF-7 cells were treated with OP. [Results] When MCF-7 cells were treated with 8 , 16 μmol/L of OP for 48 h, the percent of positive TUNEL ceils and DNA fragments increased. The percentages of apoptosis were （6.6±1.32） and （16.3±3.48）, respectively [（4.3±1.18） in control]. Fluorescence value of Bcl2, was （63.5±5.84） and （30.3±2.49）, respectively [（57.2±4.07） in control]. The Bcl2 mRNA expression significantly increased when MCF-7 cells were treated with 4μmol/L of OP. Treated with 16 μmol/L of OP, the Bcl2 mRNA expressions decreased, Bax and caspase 3 mRNA expressions increased. [Condusion] Treated with 8 and 16 μmol/L of OP, the apoptosis of MCF-7 cells and DNA fragments percentage increase, which may be caused by up-regulation of Bax and caspase 3 expression, down-regulation of bel2 expression in MCF-7 breast cancer cells.