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目的研究藻红蛋白(PE)对人乳腺癌MCF-7细胞的增殖抑制效果和诱导凋亡作用。方法采用MTT法和SRB法检测PE对人乳腺癌MCF-7细胞增殖的抑制作用。采用倒置显微镜、荧光显微镜Hoechst33258染色观察MCF-7细胞凋亡细胞形态。AnnexinV-FITC/PI双荧光染色观察细胞凋亡过程中特异的磷脂酰丝氨酸(PS)外翻情况确定凋亡的出现。流式细胞仪观察PE对MCF-7细胞周期的影响。结果PE对MCF-7细胞有较强的生长抑制作用,并且呈一定的剂量依赖性,其72h的IC50值为147.82μg/mL,GI50值为109.76μg/mL。倒置显微镜下与基本贴壁的饱满、边沿清晰的对照组细胞比较,给药组细胞有明显的细胞变粗糙,贴壁细胞减少的现象,而且浓度越高,作用时间越长,悬浮细胞越多。荧光显微镜下可见各给药组MCF-7细胞出现不同程度深染亮点的细胞核质,染色质浓集固缩并伴有凋亡小体形成。激光共聚焦显微镜下可见给药组细胞膜被染成绿色,细胞核被染成红色。流式细胞仪显示PE可阻滞MCF-7细胞从S期进入G2/M期,促使细胞凋亡。结论PE对MCF-7细胞有较强的抑制作用,可使其凋亡,其作用机制与周期阻滞有关。
Objective To study the inhibitory effect of phycoerythrin (PE) on the proliferation of human breast cancer MCF-7 cells and the induction of apoptosis. Methods The inhibitory effect of PE on the proliferation of human breast cancer MCF-7 cells was detected by MTT assay and SRB assay. The morphological changes of MCF-7 cells were observed by inverted microscope and Hoechst 33258 staining. Annexin V-FITC / PI double staining was used to observe the specific phosphatidylserine (PS) valgus during apoptosis to determine the appearance of apoptosis. The effect of PE on the cell cycle of MCF-7 cells was observed by flow cytometry. Results PE had a strong growth inhibitory effect on MCF-7 cells in a dose-dependent manner. The IC50 value at 72 h was 147.82 μg / mL and GI50 was 109.76 μg / mL. Under the inverted microscope, compared with the control group cells with full attachment and sharp edges, the cells in the treatment group had obvious cell roughening and reduced adherent cells, and the higher the concentration, the longer the action time, the more suspension cells . Fluorescence microscopy showed that the MCF-7 cells in each group showed deep-staining bright spots of nuclear cytoplasm, chromatin condensation and shrinkage accompanied by the formation of apoptotic bodies. Under confocal laser scanning microscope, the cell membrane of the administration group was dyed green and the nucleus was dyed red. Flow cytometry showed that PE could block MCF-7 cells from S phase into G2 / M phase and induce apoptosis. Conclusion PE has a strong inhibitory effect on MCF-7 cells, which can induce apoptosis. Its mechanism is related to the cycle arrest.