重组人p53腺病毒联合氟维司群对乳腺癌细胞MCF-7凋亡的影响

Effects of Ad-p53 plus fulvestrant on induced apoptosis in MCF-7 breast cancer cells

目的 探讨重组人p53腺病毒（Ad-p53）联合氟维司群对人乳腺癌细胞MCF-7凋亡的影响.方法 采用雌激素受体（ER）阳性MCF-7乳腺癌细胞为研究对象,设对照组（空载病毒组）、氟维司群组、Ad-p53组、Ad-p53＋氟维司群组作用于MCF-7细胞,处理72 h后,倒置显微镜下观察各组细胞的形态学变化;采用蛋白质印迹法（Western blot）检测各组细胞p53蛋白、ER蛋白的表达水平;流式细胞术法观察细胞的凋亡情况;MTS法观察MCF-7各组细胞增殖抑制的情况.结果 Ad-p53与氟维司群联合作用于MCF-7细胞后,细胞生长受到显著抑制,几乎无增长;流式细胞术观察到单用Ad-p53和氟维司群细胞凋亡率分别是（14.5±2.9）％、（13.3±1.2）％,联合用药组细胞凋亡率为（38.1±5.9）％,显著高于单用药组（P =0.001）;MTS法得到Ad-p53和氟维司群分别处理MCF-7细胞24、48、72、96 h后增殖抑制率分别为（8.21±0.54）％、（28.50±1.42）％、（50.14±0.78）％、（58.25±2.92）％和（9.73±1.68）％、（25.26±0.82）％、（35.25±3.94）％、（46.37±2.56）％,联合用药处理24、48、72、96 h后,细胞的增殖抑制率分别为（12.42±1.76）％、（35.20±0.58）％、（62.08±2.56）％及（75.43±3.56）％,联合用药组细胞增殖抑制率在各个时间段均高于单用Ad-p53或氟维司群组（P＜0.05）;蛋白质印迹检测表明对照组和氟维司群组p53蛋白低表达,Ad-p53组和联合用药组p53蛋白高表达,但是组间差异无统计学意义;Ad-p53组ER蛋白表达高于对照组,联合用药组ER蛋白表达明显低于单用Ad-p53组（P＜0.05）.结论 氟维司群联合重组人p53腺病毒可显著促进乳腺癌细胞凋亡,抑制细胞增殖,二者可能具有协同作用.

Objective To explore the apoptosis-inducing effect of Ad-p53 plus fulvestrant in MCF-7 breast cancer cells.Methods ER-positive MCF-7 breast cancer cells were used.MCF-7 cells were established with control,fulvestrant alone,Ad-p53 alone and Ad-p53 plus fulvestrant.The morphological changes of MCF-7 after drug combination were observed by inverted microscopy.Methyltetrazolium salt （MTS） assay was used to determine the proliferation of MCF-7 cells.The protein expressions of p53 and ER were examined by Western blot.Flow cytometry was used to detect apoptosis.Results Significant inhibition of cell growth was observed by inverted microscopy.Flow cytometry revealed that the apoptosis rate of using fulvestrant alone or Ad-p53 alone was（13.3 ± 1.2）% and（14.5 ± 2.9）%.The apoptosis rate of combined group was （38.1 ± 5.9） % and it was higher than Ad-p53 or fulvestrant alone （P =0.001,P =0.001）.MTS assay indicated that,after treating for 24,48,72,96 h by Ad-p53 alone or fulvestrant alone,the inhibition of cell proliferation rate was （8.21 ± 0.54） %,（28.50 ± 1.42） %,（50.14 ± 0.78） %,（58.25 ±2.92）% and （9.73 ± 1.68）%,（25.26 ±0.82）%,（35.25 ±3.94）% and （46.37 ±2.56）% respectively.Simultaneously,the inhibition of cell proliferation rate of combined group was （12.42 ± 1.76）%,（35.20 ±0.58）%,（62.08 ±2.56）% and （75.43 ±3.56）%.It further confirmed the increase of cell proliferation inhibition rate after drug combination （P 〈 0.05）.Western blot showed that the p53 protein expression level increased obviously with Ad-p53 in MCF-7cells.Ad-p53 increased the level of ER expression while drug combination reduced the level of ER expression.Conclusions Ad-p53 plus fulvestrant can significantly inhibit cell growth and induce cell apoptosis.And there is probably some synergistic effects between both.