多西他赛对乳腺癌细胞中Cav-1表达及细胞增殖的影响

目的 研究多西他赛对乳腺癌细胞中Cav-1(Cav-1)表达及细胞增殖的影响.方法 通过Real-time PCR和Western-blot检测不同浓度多西他赛作用下乳腺癌细胞MCF-7中Cav-1的表达情况;野生型MCF-7细胞株为对照组,通过基因转染技术建立Cav-1 过表达的MCF-7细胞株(转染组)及空载体的MCF-7细胞株(空载体组),利用CCK-8比色法分析多西他赛对各组细胞增殖的影响.通过Western-blot分别检测多西他赛作用下各组细胞中凋亡相关蛋白Bax和Bcl-2的表达情况以及PI3K/AKT 信号转导通路的活化情况.结果 在5~40 μg/mL浓度范围内,随着多西他赛作用浓度的增大,MCF-7细胞中Cav-1的蛋白表达量逐渐增加.不同浓度多西他赛对MCF-7细胞的增殖均有明显的抑制作用;在同一浓度多西他赛作用下,转染Cav-1组细胞的增殖抑制率明显高于对照组和空载体组(P<0.01).与对照组和空载体组相比,20 μg/mL多西他赛作用后转染组细胞中Bax的表达升高(P<0.01),而Bcl-2的表达降低(P<0.01),并且磷酸化AKT (p-AKT)蛋白的表达水平下降(P<0.01),而对照组和空载体组之间Bax、Bcl-2以及p-AKT蛋白的表达则无明显差异(P>0.05).结论 多西他赛可以通过促进Cav-1的表达影响PI3K/AKT 信号转导通路,进而调节凋亡相关蛋白的表达抑制MCF-7 细胞的增殖.

Effect and potential mechanism of Cav-1 in Docetaxel inhibiting breast cancer MCF-7 cell proliferation

Objective To investigate the effect and potential mechanism of Cav-1 (Cav-1) on the proliferation inhibiting of breast cancer MCF-7 cell by Docetaxel.Methods The transfection group was established by stable transfection of pcDNA3.1-Cav-1 into MCF-7 cells, while the vector group (pcDNA3.1 vector transfected) and control group (non-transfection MCF-7 cells) were used as control.The Cav-1 expression of MCF-7 cells treated by Docetaxel was detected by Real-time PCR and Western-blot.CCK-8 assay was used to analyze the cell proliferation.The expression of Bax and Bcl-2 proteins and the activation of PI3K/AKT signal pathway were examined by Western-blot.Results The expression of Cav-1 in MCF-7 cells was gradually enhanced with the increasing of Docetaxel concentration in the range of 5-40 μg/mL.The inhibitory rate of cell proliferation in Cav-1 transfection group was significantly higher than that of the control groups after Docetaxel treatment (P<0.01).Compared with the control groups, Bax expression increased and Bcl-2 and p-AKT expression decreased in Cav-1 transfection group (P<0.01), while there was no significant difference between the two control groups (P>0.05).Conslusion Docetaxel might inhibit the proliferation of MCF-7 cells by promoting the Cav-1 expression, which can regulate the apoptosis-related protein expression and suppress the activation of PI3K/AKT signaling pathway.