碧萝芷对三阴性乳腺癌MDA-MB-231细胞增殖和凋亡及转移影响  

Effect of pycnogenol on proliferation,apoptosis and metastasis of human breast cancer MDA-MB-231cells

作　　者：杨春姝 秦光远 罗小萌[2] 徐竞忆 杨娉婷[2] 郑新宇[1] YANG Chun-shu;QIN Guang-yuan;LUO Xiao-meng;XU Jing-yi;YANG Ping-ting;ZHENG Xin-yu(First Affiliated Hospital,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,辽宁沈阳110001 [2]中国医科大学附属第一医院,风湿免疫科,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2022年第21期1528-1535,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：辽宁省科学技术厅2021“兴辽英才计划”(XLYC2002062)。

摘　　要：目的探讨松树皮提取物碧萝芷对乳腺癌MDA-MB-231细胞增殖、凋亡和转移的影响及可能机制。方法分别用30、60和90mg/L的碧萝芷和RPMI 1640培养基(对照组)处理乳腺癌MDA-MB-231细胞,采用MTT法检测碧萝芷对乳腺癌MDA-MB-231细胞增殖抑制作用;克隆形成实验检测对MDA-MB-231细胞体外生长能力的影响;流式细胞仪检测细胞凋亡率;Transwell小室法分别检测各浓度碧萝芷对乳腺癌MDA-MB-231细胞迁移和侵袭能力的影响;蛋白质印迹法分别检测碧萝芷处理后,乳腺癌MDA-MB-231细胞凋亡相关蛋白Bcl-2、Bax、Caspase-9、Caspase-3、PARP以及基质金属蛋白酶(MMP)-2、MMP-9的表达;明胶酶谱法检测MMP-2和MMP-9的活性。结果MTT结果显示,与对照组相比,碧萝芷呈剂量依赖及时间依赖性抑制乳腺癌MDA-MB-231细胞增殖,差异有统计学意义,F浓度=178.335,P浓度<0.001;F时间=3.476,P时间=0.035;二因素间交互作用:F时间×浓度=3.162,P时间×浓度=0.007,为协同作用。克隆形成实验结果显示,对照组及30、60、90mg/L碧萝芷组克隆形成率分别为(20.09±1.96)%、(14.34±2.05)%、(12.90±1.37)%和(9.81±1.27)%,差异有统计学意义,F=57.902,P<0.001。流式细胞仪检测结果显示,对照组及30、60、90mg/L碧萝芷组的细胞凋亡率分别为(3.58±0.26)%、(5.75±0.40)%、(9.60±0.20)%和(19.71±0.38)%,差异有统计学意义,F=4507.561,P<0.001。Transwell小室结果显示,对照组及30、60、90mg/L碧萝芷组的迁移细胞数目分别为41±3、36±4、22±5和9±2,差异有统计学意义,F=131.882,P<0.001;侵袭转移的细胞数目分别为30±4、27±3、17±2和10±2,差异有统计学意义,F=89.964,P<0.001。蛋白质印迹法结果显示,碧萝芷可上调Bax(F=973.558,P<0.001)、Caspase-9(F=679.196,P<0.001)、Caspase-3(F=912.678,P<0.001)和PARP(F=70.155,P<0.001)蛋白表达,下调Bcl-2(F=216.980,P<0.001)蛋白表达。同时,各组MDA-MB-231细胞中的MMP-2(F=160.844,P<0.001)及MMP-9(F=94.135,P<0.001)蛋白表达水�Objective To investigate the effect and possible mechanisms of Pycnogenol on proliferation,apoptosis,and metastasis of human breast cancer MDA-MB-231cells.Methods Breast cancer MDA-MB-231cells were treated with 30,60,90mg/L Pycnogenol and RPMI 1640culture medium respectively.MTT tests were used to identify Pycnogenol's inhibitory effect on the proliferation of breast cancer MDA-MB-231cells;Clonogenic assay was used to detect the effect of Pycnogenol on the growth of breast cancer MDA-MB-231cells in vitro.The apoptosis rate was measured by flow cytometry.The effects of Pycnogenol at different concentrations on the migration and invasion of breast cancer MDA-MB-231cells were detected by Transwell chamber assay.The expression of apoptosis-related proteins Bcl-2,Bax,Caspase-9,Caspase-3,and PARP in MDA-MB-231cells treated by Pycnogenol was determined by Western blot analysis.The expression of matrix metalloprotein(MMP)-2and MMP-9was also determined by Western blot analysis.The activities of MMP-2and MMP-9were measured by Gelatinize assay.Results The results of MTT assay showed that compared with the control group,Pycnogenol inhibited the proliferation of MDA-MB-231cells in a dose-dependent and time-dependent manner,with a statistically significant difference(Fconcentration=178.335,Pconcentration<0.001;Ftime=3.476,Ptime=0.035;two factors were interacted with each other:Ftime×concentration=3.162,Ptime×concentration=0.007,which was a synergistic effect).The results of the clone formation experiment showed that the clone formation rates of the control group and 30,60,and 90mg/L Pycnogenol groups were(20.09±1.96)%,(14.34±2.05)%,(12.90±1.37)%,and(9.81±1.27)%,respectively.The difference was statistically significant(F=57.902,P<0.001).The results of flow cytometry showed that the apoptosis rates of control group and 30,60,and 90mg/L Pycnogenol groups were(3.58±0.26)%,(5.75±0.40)%,(9.60±0.20)%,and(19.71±0.38)%,respectively,indicating a significant difference(F=4507.561,P<0.001).The results of the Transwell chamber sh

关 键 词：乳腺癌 碧萝芷 增殖 凋亡 转移 

分 类 号：R737.9[医药卫生—肿瘤]