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裸花紫珠提取物的抗乳腺癌转移作用及其机制
引用本文:陈斌,罗跃华,王珊,吕红,麻俊超,刘婷,柯小琴,付剑江.裸花紫珠提取物的抗乳腺癌转移作用及其机制[J].中国实验方剂学杂志,2015,21(18):94-98.
作者姓名:陈斌  罗跃华  王珊  吕红  麻俊超  刘婷  柯小琴  付剑江
作者单位:江西中医药大学 药学院, 南昌 330004,江西省药品检验检测研究院, 南昌 330029,江西中医药大学 药学院, 南昌 330004,江西中医药大学 网络与教育技术中心, 南昌 330004,江西中医药大学 药学院, 南昌 330004,江西中医药大学 药学院, 南昌 330004,江西中医药大学 药学院, 南昌 330004,江西中医药大学 药学院, 南昌 330004;江西省药品检验检测研究院, 南昌 330029
基金项目:国家自然科学基金项目(81160530,81260656,81373955,81560639);江西省自然科学基金项目(2010GQY0147);教育部重点科技项目(211091)
摘    要:目的:研究裸花紫珠乙酸乙酯提取物(ethyl acetate extracts from Callicarpa nudiflora,ECN)的抗乳腺癌转移作用及其作用机制。方法:采用尾静脉注射肿瘤细胞法建立裸鼠实验性肺转移模型,24 h后将实验动物随机分为空白组,ECN低、中、高剂量组(0.125,0.25,0.5 g·kg-1),空白组给予等量溶剂,每日ig给药1次,连续8周。给药结束后,采用巢式PCR法检测裸鼠肺组织中人细胞角蛋白19(CK19)基因的表达,以确定肿瘤细胞的肺转移程度(n=10)。建立肿瘤细胞体外迁移、侵袭模型,以100,200 mg·L-1ECN以及10 nmol·L-1BB-94(阳性对照)处理MDA-MB-231细胞14 h(n=4),观察ECN对乳腺癌细胞系MDA-MB-231细胞的体外迁移、侵袭的影响;建立肿瘤细胞体外黏附模型,以100,200 mg·L-1ECN以及10 nmol·L-1BB-94处理MDA-MB-231细胞60 min(n=3),观察ECN对乳腺癌细胞系MDA-MB-231细胞的体外黏附能力的影响;采用Western blot法,检测MDA-MB-231细胞分别经100,200 mg·L-1ECN处理24 h后p-锌指转录因子(p-Snail)和上皮性钙黏附蛋白(Ecadherin)的表达情况。结果:体内研究表明,ECN可显著降低裸鼠肺组织中人CK19 mRNA的表达(P0.01);体外迁移实验发现,ECN 100,200 mg·L-1组的迁移细胞数与空白组相比显著减少(P0.01);体外侵袭实验显示,ECN低、高剂量组的侵袭细胞数与空白组相比显著减少(P0.01);体外黏附实验表明,ECN可干扰肿瘤细胞与细胞外基质蛋白之间的黏附(P0.01);Western blot实验结果表明,ECN可显著抑制p-Snail,并增加E-cadherin的表达。结论:ECN具有明显的抗乳腺癌转移作用。抑制细胞内p-Snail的活化可能是抗转移作用的机制之一。

关 键 词:裸花紫珠乙酸乙酯提取物  转移  乳腺癌  锌指转录因子  上皮性钙黏附蛋白
收稿时间:2015/1/21 0:00:00

Anti-metastatic Effects of Extracts from Callicarpa nudiflora via Downregulating Snail Activation on Breast Cancer Cell
CHEN Bin,LUO Yue-hu,WANG Shan,LYU Hong,MA Jun-chao,LIU Ting,KE Xiao-qin and FU Jian-jiang.Anti-metastatic Effects of Extracts from Callicarpa nudiflora via Downregulating Snail Activation on Breast Cancer Cell[J].China Journal of Experimental Traditional Medical Formulae,2015,21(18):94-98.
Authors:CHEN Bin  LUO Yue-hu  WANG Shan  LYU Hong  MA Jun-chao  LIU Ting  KE Xiao-qin and FU Jian-jiang
Institution:School of Pharmacy, Jiangxi University of Traditional Chinese Medicine (TCM), Nanchang 330004, China,Jiangxi Institute for Drug Control, Nanchang 330029, China,School of Pharmacy, Jiangxi University of Traditional Chinese Medicine (TCM), Nanchang 330004, China,Network and Education Technology Center, Jiangxi University of TCM, Nanchang 330004, China,School of Pharmacy, Jiangxi University of Traditional Chinese Medicine (TCM), Nanchang 330004, China,School of Pharmacy, Jiangxi University of Traditional Chinese Medicine (TCM), Nanchang 330004, China,School of Pharmacy, Jiangxi University of Traditional Chinese Medicine (TCM), Nanchang 330004, China and School of Pharmacy, Jiangxi University of Traditional Chinese Medicine (TCM), Nanchang 330004, China;Jiangxi Institute for Drug Control, Nanchang 330029, China
Abstract:Objective: To investigate the anti-metastatic effects of ethyl acetate extracts from Callicarpa nudiflora (ECN) and its mechanisms. Method: BALB/c Nu mice were used to establish in vivo experimental metastasis mode and the mice were randomly divided into control group and ECN groups treated with 0.125, 0.25, 0.5 g· kg-1 ECN respectively. After 8 weeks of gavage (once a day), human cytokeratin 19(CK19) mRNA expression in lungs of mice was checked by nest PCR to determine effects of ECN on the formation of lung metastatic foci of breast cancer. In vitro invasion and migration assay were involved to determine effects of ECN on metastatic properties of breast cancer cells. MDA-MB-231 cells were treated using ECN at dose of 100, 200 mg· L-1 and BB-94 with the final concentration of 10 nmol· L-1(n=4). After 14 h incubation, the invaded and migrated cells were calculated. In vitro adhesion assay was also used in this study. MDA-MB-231 cells were treated using ECN at dose of 100, 200 mg· L-1and BB-94 with the final concentration of 10 nmol· L-1(n=3). After 60 min incubation, cells attached to matrix proteins were detected. p-Snail and E-cadherin expressions were checked by western blot method in current study to detect the effects of ECN on activation of Snail in MDA-MB-231 cells, which were treated with 100, 200 mg· L-1ECN for 24 h. Result: ECN induced a significant decrease on human CK19 mRNA expression in lung of tumor-bearing mice(P<0.01). Moreover, in vitro adhesion, migration and invasion of MDA-MB-231 cells were inhibited significantly when these cells incubated with ECN (P<0.01). It was also showed that ECN affected activity of Snail and expression of E-cadherin in breast cancer cells. Conclusion: ECN shows significant anti-metastatic properties. The mechanisms of action of ECN may be associated with suppression of Snail activation.
Keywords:Callicarpa nudiflora  metastatisis  breast cancer  Snail  E-cadherin
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