多西他赛对乳腺癌细胞中Cav-1表达及细胞增殖的影响

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

关键词:  Cav-1  多西他赛  乳腺癌细胞  细胞增殖  基因治疗

Key words:  Cav-1  Docetaxel  breast cancer cells  cell proliferation  gene therapy

摘要:

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。

Abstract:

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.