左归丸抗乳腺癌转移及其作用机制  

Anti-metastatic Effect of Zuoguiwan on Breast Cancer Cells and Its Mechanisms of Action

作　　者：邹乐兰 刘红宁[2] 吕红 徐焕军 朱小翠[4] 张舟妙 付剑江[4] 

机构地区：[1]南昌大学医院,南昌330031 [2]江西中医药大学中医基础理论分化发展研究中心,南昌330004 [3]江西中医药大学网络与教育技术中心,南昌330004 [4]江西中医药大学药学院,南昌330004

出　　处：《中国实验方剂学杂志》2017年第23期85-90,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81560639;81660680);江西省自然科学基金项目(20171BAB205097)

摘　　要：目的:观察左归丸的抗乳腺癌转移作用,探讨其作用机制。方法:采用股骨内注射法建立人乳腺癌细胞MDAMB-231骨转移裸鼠模型,受试裸鼠随机分为空白组(蒸馏水),左归丸低、高剂量(21,42 g·kg-1)组。连续给药8周后,采用巢式聚合酶链式反应(PCR)检测裸鼠骨髓组织中人细胞角蛋白(ck19)基因的表达,确定肿瘤转移情况;采用Oris法,检测左归丸含药血清对乳腺癌细胞MDA-MB-231体外迁移能力的影响;采用重组基底膜侵袭法,观察左归丸含药血清的体外抗侵袭作用;采用Alexa Fluor488标记的鬼笔环肽(Phalloidin)进行荧光染色,观察左归丸含药血清对乳腺癌细胞纤维型肌动蛋白(F-actin)聚合的影响;采用蛋白质印迹法(Western blot)检测左归丸含药血清对MDA-MB-231细胞甲状旁腺激素相关蛋白(PTHr P)和4型趋化因子受体(CXCR4)表达的影响。结果:与空白组比较,左归丸高、低剂量均可显著抑制小鼠骨髓组织中人ck19基因表达(P<0.01)。Oris迁移表明,左归丸含药血清可显著抑制转化生长因子-β(TGF-β)诱导的MDA-MB-231细胞迁移;重组基底膜侵袭实验显示,左归丸含药血清也可显著抑制乳腺癌细胞对重组基底膜的侵袭能力。给药组侵袭细胞数较空白组显著降低(P<0.01)。Phalloidin荧光染色结果显示,左归丸含药血清可显著抑制F-actin聚合,抑制F-actin聚合体的形成。Western blot结果表明,左归丸含药血清可显著抑制乳腺癌细胞PTHr P,CXCR4蛋白表达。结论:左归丸具有明显的抗乳腺癌骨转移作用,其机制与抑制肿瘤细胞PTHr P,CXCR4表达,从而直接抑制其体外运动活性相关。Objective： To investigate the anti-metastatic effect of Zuoguiwan and its mechanisms of action. Method： The bone marrow metastasis mode of nude mice was established by intra-femoral injection. The tumor-bearing mice were randomly divided into three groups： control group, low-dose Zuoguiwan （21 g·kg^-1 per day） treatment group and high-dose Zuoguiwan （42 g·kg^-1 per day） treatment group. After successive administration for 8 weeks, the expression of human ck19 gene was detected by nest polymerase chain reaction （PCR） assay to determine the metastatic foci of MDA-MB-231 cells in bone marrow. Oris migration assay was performed to determine the effect of Zuoguiwan-containing sera on the migration of MDA-MB-231 ceils. Reconstituted basement membrane invasion assay was conducted to decide the effect of Zuoguiwan-containing sera on the invasion of MDA-MB-231 cells. F-actin polymerization was detected with Alexa Fluor 488 phalloidin and fluorescence microscopy. The effect of Zuoguiwan-containing sera on parathyroid hormone-related protein （PTHrP） and C-X-C chemokine receptor type 4 （CXCR4） expressions in MDA-MB-231 was determined by Western blot assay. Result： Compared with control group, both low-dose Zuoguiwan group and high-dose Zuoguiwan group exhibited a significant suppression on the expression of ck19 gene in bone marrow of tumor-bearing mice （P 〈 0.01 ）. Data from Otis migratory assay showed that Zuoguiwan-containing sera inhibited transforming growth factor- beta （TGF-β） -induced migration in MDA-MB-231 cells significantly. It was also found that the invasive ability of MDA-MB-231 ceils through reconstituted basement membrane was inhibited by Zuoguiwan-containing sera in reconstituted basement membrane invasion assay. The amount of invaded cells in Zuoguiwan-containing sera treatment groups was less than that in control group obviously （P 〈 0.01 ）. Data from fluorescence assay showed that Zuoguiwan-containing sera inhibited F-actin polymerization of breast cancer ce

关 键 词：左归丸 破骨性骨转移 乳腺癌 甲状旁腺激素相关蛋白 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]