半边旗提取物5F对乳腺癌细胞内活性氧水平的影响Effects of 5F from Pteris semiinnata L. on Reactive Oxygen Species Levels in Breast Cancer Cells
邱万寿,刘威,吴珏堃,李玺,龙梅珺
QIU Wanshou,LIU Wei,WU Juekun,LI Xi,LONG Meijun
摘要(Abstract)：
目的观察中草药半边旗提取物5F(Ent-11a-hydroxy-15-oxo-kaur-16-en-19-oic-acid,5F)对乳腺癌细胞的抑制作用及其对乳腺癌细胞内活性氧(reactive oxygen species,ROS)水平的影响。方法不同浓度(0、5、10、20、40μg/m L)的5F分别处理3种不同乳腺癌细胞(MCF-7、MDA-MB-231以及SK-BR-3),用CCK-8法检测5F对不同乳腺癌细胞株增殖的影响;流式细胞仪检测5F处理后的各乳腺癌细胞株的凋亡率;检测不同浓度的5F处理后的乳腺癌细胞内ROS的水平。结果 5F可抑制MDA-MB-231、MCF-7、SK-BR-3三种乳腺癌细胞的增殖,其抑制效果与5F的浓度及作用时间呈显著正相关(P<0.05);经5F作用24 h后,MDA-MB-231细胞、MCF-7细胞及SK-BR-3细胞的凋亡率较阴性对照组明显增加(P<0.05);5F作用于乳腺癌细胞株,ROS相对含量均较未处理组降低,且随5F浓度升高ROS水平降低越明显(P<0.05)。结论 5F可抑制乳腺癌细胞的增殖,其抑制效果与5F浓度和作用时间相关;5F能降低细胞内ROS的生成水平,并呈现剂量依赖性。
OBJECTIVE To explore the anti-cancer effects and effects on reactive oxygen species levels of ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid( 5F) in human breast cancer cells. METHODS Human breast cancer cells( MCF-7,MDA-MB-231,SKBR-3) were treated with 5F in different concentrations( 0,5,10,20,40 μg / m L) for different periods respectively. Cell viability was detected by a cell counting kit-8. The appotosis rates of cells were examined by flow cytometer. To detect the level of ROS in breast cancer cells after treated with different concentrations of 5F. RESULTS 5F inhibited the proliferation of MDA-MB-231,MCF-7,SKBR-3 in a concentration- and time-dependent manner( P < 0. 05). After treated by 5F for 24 h,the apoptosis rate of MDA-MB-231,MCF-7,SK-BR-3 were increased significantly when compared with the control group( P < 0. 05). The ROS level of 5F in patients of the treated group were lower than that of the untreated groups,and in a concentration-dependent manner( P < 0. 05). CONCLUSION5 F could inhibit the proliferation of breast cancer cells in a concentration- and time-dependent manner. 5F can reduce intracellular ROS levels in a concentration-dependent manner.
关键词(KeyWords)：
半边旗;5F;乳腺癌细胞;活性氧
pteris semipinnata L;5F;breast cancer cell;reactive oxygen species
基金项目(Foundation): 广东省女性肿瘤用药研究基金项目(2014D09)
作者(Author):
邱万寿,刘威,吴珏堃,李玺,龙梅珺
QIU Wanshou,LIU Wei,WU Juekun,LI Xi,LONG Meijun
参考文献(References)：
- [1]陈建发,陈引香,李萍,等.半边旗提取物5F对人胃癌细胞凋亡的影响及其机制[J].南方医科大学学报,2011,31(8):1345-1348.
- [2]Chen GG,Leung J,Liang NC,et al.Ent-11α-hydroxy-15-oxokaur-16-en-19-oic-acid inhibits hepatocellular carcinoma in vitro and in vivo via stabilizing Ik Bα[J].Invest New Drugs,2012,30:2210-2218.
- [3]吴科锋,吕应年,刘义,等.半边旗提取物5F-Na盐注射液制备及其对小鼠Sis0型肉瘤抑制作用研究[J].时珍国医国药,2011,22(3):546-549.
- [4]兰柳波,梁念慈,莫丽儿,等.半边旗抗肿瘤有效成分5F的纯化及其体外增效作用[J].中国药理学通报,2003,19(7):804-807.
- [5]Lafon C,Mathieu C,Guerrin M,et al.Transforming growth factor beta 1-induced apoptosis in human ovarian carcinoma cells:protection by the antioxidant N-acetylcysteine and bcl-2[J].Cell Growth Differ,1996,7:1095-1104.
- [6]Bladier C,Wolvetang EJ,Hutchinson P,et al.Response of a primary human fibroblast cell line to H2O2:senescence-like growth arrest or apoptosis[J]?Cell Growth Differ,1997,8:589-598.
- [7]Chen GG,Liang NC,Lee JF,et al.Over-expression of Bcl-2 against Pteris semipinnata L-induced apoptosis of human colon cancer cells via a NF-kappa B-related pathway[J].Apoptosis,2004,9:619-627.
- [8]Muindi JR,Sinha BK,Gianni L,et al.Hydroxyl radical production and DNA damage induced by anthracycline-iron complex[J].FEBS Lett,1984,172:226-230.
- [9]Myers CE,Mc Guire W,Young R.Adriamycin:amelioration of toxicity by alpha-tocopherol[J].Cancer Treat Rep,1976,60:961-962.
- [10]Berthiaume JM,Wallace KB.Adriamycin-induced oxidative mitochondrial cardiotoxicity[J].Cell Biol Toxicol,2007,23:15-25.
- [11]Chen Y,Daosukho C,Opii WO,et al.Redox proteomic identifi-cation of oxidized cardiac proteins in adriamycin-treated mice[J].Free Radic Biol Med,2006,41:1470-1477.
- [12]Shioji K,Kishimoto C,Nakamura H,et al.Overexpression of thioredoxin-1 in transgenic mice attenuates adriamycin-induced cardiotoxicity[J].Circulation,2002,106:1403-1409.
- [13]Gao J,Xiong Y,Ho YS,et al.Glutathione peroxidase 1-deficient mice are more susceptible to doxorubicin-induced cardiotoxicity[J].Biochim Biophys Acta,2008,1783:2020-2029.
- [14]Cai Y,Lu J,Miao Z,et al.Reactive oxygen species contribute to cell killing and P-glycoprotein downregulation by salvicine in multidrug resistant K562/A02 cells[J].Cancer Biol.Ther,2007,6:1794-1799.
- [15]Joshi G,Hardas S,Sultana R,et al.Glutathione elevation by gamma-glutamyl cysteine ethyl ester as a potentialtherapeutic strategy for preventing oxidative stress in brain mediated by in vivo administration of adriamycin:implication for chemobrain[J].J Neurosci Res,2007,85:497-503.