原花青素通过Caspase途径诱导乳腺癌MCF-7细胞凋亡

Procyanidin induces MCF-7 cell apoptosis by Caspase way

目的研究原花青素(Proanthocyanidin/Procyanidin,PC)诱导人乳腺癌细胞凋亡以及凋亡通路,揭示其抗乳腺癌的部分作用机制。方法常规培养细胞,24 h后随机分为阳性对照组、空白对照组和PC组,MTT法检测PC对人乳腺癌细胞增殖的抑制作用;流式细胞术检测凋亡率和细胞周期;ELISA法检测培养上清液中Caspase-9、Caspase-12的含量。结果 PC(10～320μg/mL)6个剂量组可显著抑制MCF-7细胞的增殖,且呈时间和剂量依赖性;在40、80和160μg/mL 3个浓度下体外培养,PC可提高MCF-7细胞的凋亡率,并诱导出现G2-M细胞周期阻滞;PC作用48 h后,Caspase-9、Caspase-12蛋白表达量增加。结论 PC抑制人乳腺癌MCF-7细胞的作用机制与阻滞细胞周期于G2-M期和诱导细胞凋亡有关,其凋亡通路与Caspase途径有关,并涉及Caspase家族的内质网和线粒体两条通路。

Objective To investigate the mechanism that procyanidin induced apoptosis of human breast canc-er cells and the apoptosis pathway, and to reveal part of the mechanism of its anti-breast cancer. Methods Cells were cultivated in vitro, some good cells were taken and divided into positive control group, negative control group and PC group randomly after cultured for 24 h. The cytostatic effect of procyanidin on human breast cancer cell line MCF-7 was evaluated by MTT assay. Flow cytometry was utilized for measuring the apoptosis and cell cycle. The expressions of Caspase-9,Caspase-12 in cultured supernatant were detected by ELISA assay. Results Procyanidin inhibited cell growth in a concentration-and time-dependent manner. At the dosage of 40,80 and 160 μg/mL in cultured cells, the ap-optosis rates was elevated,and G2-M phrase was arrested. Significant up-regulation of Caspase-9, Caspase-12 was ob-served in MCF-7 cells 48 h after treatment with procyanidin. Conclusion Procyanidin inhibited the proliferation of MCF-7 ceils associated with cycle arrest in G2-M phase,induction of apoptosis. Caspase pathway was related in the ap-optosis pathway,moreover,endoplasmic reticulum and mitochondria were involved in the Caspase family.