碧萝芷对乳腺癌MCF-7细胞增殖、迁移和转移的影响  

Effect of pycnogenol on proliferation, migration and metastasis of human breast cancer MCF-7 cells

作　　者：杨春姝 秦光远 郑新宇[1,2] Yang Chunshu;Qin Guangyuan;Zheng Xinyu(Laboratory No. 1 of Cancer Institute ,First Affiliated Hospital of China Medical University, Shenyang 110001, China;Department of Breast Surgery, First Affiliated Hospital of China Medical University, Shenyang 110001, China)

机构地区：[1]中国医科大学附属第一医院肿瘤研究所第一研究室,沈阳110001 [2]中国医科大学附属第一医院肿瘤研究所乳腺外科,沈阳110001

出　　处：《中华乳腺病杂志（电子版）》2019年第3期165-172,共8页Chinese Journal of Breast Disease(Electronic Edition)

摘　　要：目的探讨抗氧化剂碧萝芷对乳腺癌MCF-7细胞的抑制作用,并对观察碧萝芷对MCF-7细胞的转移和迁移功能的影响。方法用10、20、40μg/ml的碧萝芷处理乳腺癌MCF-7细胞作为实验组,空白对照组只加入DMEM培养基(无血清), MTT法在处理细胞后24、48、72 h于波长490 nm处测定各组吸光度值;用流式细胞仪检测细胞凋亡率;用β-半乳糖苷酶染色法检测MCF-7细胞衰老率;用Transwell小室法分别检测各浓度碧萝芷对乳腺癌MCF-7细胞迁移和侵袭能力的影响;用Western blot法分别检测乳腺癌MCF-7细胞衰老相关蛋白P53、P21、P16、P27、E2F1以及基质金属蛋白酶(matrix metalloproteinase, MMP)-2、MMP-9的表达。细胞迁移、侵袭数目及各种蛋白的表达水平比较采用单因素方差分析,吸光度值比较采用重复测量的方差分析,两两比较采用LSD法。结果 MTT结果显示,在处理乳腺癌MCF-7细胞24、48、72 h后,各组吸光度值比较,差异有统计学意义(F=149.439,P<0.001),在不同时间点之间比较,差异有统计学意义(F=27.922,P<0.001);分组与时间点之间存在交互作用(F=18.466,P<0.001)。流式细胞仪检测结果显示:空白对照组及10、20、40μg/ml碧萝芷组的细胞凋亡率分别为(2.36±0.27)%、(6.44±1.43)%、(7.52±2.09)%和(11.68±1.65)%,差异有统计学意义(F=19.143,P<0.001)。β-半乳糖苷酶染色法结果显示:空白对照组和10、20、40μg/ml碧萝芷组的细胞衰老率分别为(5.35±1.32)%、(20.08±2.14)%、(40.55±4.61)%和(59.26±4.10)%,差异有统计学意义(F=150.150,P<0.001)。Transwell小室结果显示:空白对照组及10、20、40μg/ml碧萝芷组的迁移细胞数目分别为(41±5)、(27±2)、(19±1)、(11±2)个,差异有统计学意义(F=59.330,P<0.001);侵袭转移的细胞数目分别为(24±4)、(17±1)、(12±2)、(7±2)个,差异也有统计学意义(F=26.230,P<0.001)。Western blot结果显示:碧萝芷可上调P53、P21、P16、P27蛋白表达(F=263.905、424.937、217.515、391Objective To investigate the inhibitory effect of the antioxidant pycnogenol on human breast cancer MCF-7 cells, and to observe the effect of pycnogenol on the metastasis and migration of MCF-7 cells.Methods MCF-7 cells were treated with 10, 20, 40 μg/ml pycnogenol, respectively, as experimental groups. The control group was only treated with blank DMEM medium(without fetal bovine serum). The optical density of MCF-7 cells at the wavelength of 490 nm was determined by the MTT method at 24, 48 and 72 h after treatment. The apoptosis rate of MCF-7 cells was measured by the flow cytometry. The senescence rate of MCF-7 cells was measured by β-galactosidase staining. The effect of pycnogenol at different concentrations on the migration and invasion of MCF-7 cells were detected by Transwell chamber assay. The expression of senescence-associated proteins P53, P21, P16, P27 and E2 F1 in MCF-7 cells was determined by Western blot analysis. The expression of matrix metalloproteinase(MMP)-2 and MMP-9 was also determined by Western blot analysis. The number of migrated cells and metastatic cells, and the protein expression among four groups were compared by one-factor analysis of variance. The optical density was compared by repeated measurement analysis of variance and pairwise comparison was performed by LSD method.Results The result of MTT assay showed that at 24, 48, and 72 h after treatment, the optical density of breast cancer MCF-7 cells was significantly different among four groups(F=149.439, P<0.001) and at different time points(F=27.922,P<0.001);there was an interaction between grouping and time points(F=18.466, P<0.001). The result of flow cytometry showed that the apoptosis rate of MCF-7 cells was(2.36±0.27)%,(6.44±1.43)%,(7.52±2.09)% and(11.68±1.65)% in the blank control group, 10, 20 and 40 μg/ml pycnogenol group, respectively, indicating a significant difference(F=19.143, P<0.001). The result of β-galactosidase staining showed that the senescence rate of MCF-7 cells was(5.35±1.32)%,(20.08±2.14)%,(40.

关 键 词：乳腺肿瘤 细胞衰老 迁移 转移 碧萝芷 

分 类 号：R737.9[医药卫生—肿瘤]