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本文研究了人乳腺癌细胞MCF-7在滚瓶培养形成大量细胞聚集体时,猪软骨多糖对MCF-7的诱导凋亡作用,并初步研究了其凋亡前后的差异表达蛋白。采用Hoechst 33342/PI染色于共聚焦显微镜下观察细胞的凋亡特征;Annexin V-FITC/PI检测细胞凋亡;流式细胞仪检测细胞周期的变化;并提取MCF-7细胞的全蛋白,利用二维凝胶电泳(2-DE)结合MALDI-TOF质谱技术检测差异表达蛋白。结果表明:MCF-7细胞在猪软骨多糖作用后出现典型的凋亡现象,通过流式细胞仪检测发现在猪软骨多糖作用12 h,24 h的MCF-7凋亡率分别为20.23%和44.7%;同时发现MCF-7细胞在猪软骨多糖作用后,细胞周期被阻滞在了S期;初步鉴定出2个差异表达蛋白,其中一个为肌动蛋白actin,另一个为抗氧化类蛋白PRDX6。
In this study, we investigated the apoptosis of MCF-7 induced by porcine cartilage polysaccharide (MCF-7) when human breast cancer cell line MCF-7 was cultured in roller bottles and the differentially expressed proteins before and after apoptosis were studied. Cell apoptosis was observed by confocal microscopy with Hoechst 33342 / PI staining. Apoptosis was detected by Annexin V-FITC / PI. Flow cytometry was used to detect cell cycle changes. Total protein was extracted from MCF-7 cells. Two-dimensional gel electrophoresis (2-DE) combined with MALDI-TOF mass spectrometry detection of differentially expressed proteins. The results showed that MCF-7 cells showed a typical apoptosis phenomenon after pig cartilage polysaccharide treatment. The apoptosis rate of MCF-7 cells was 20.23% and 44.7 %. At the same time, it was found that MCF-7 cells were arrested in S phase after pig cartilage polysaccharide treatment. Two differentially expressed proteins were identified, one of which was actin and the other was anti-oxidation protein PRDX6 .