Brucine suppresses breast cancer metastasis via inhibiting epithelial mesenchymal transition and matrix metalloproteinases expressions
Brucine suppresses breast cancer metastasis via inhibiting epithelial mesenchymal transition and matrix metalloproteinases expressions
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DOI：
10.1007/s11655-017-2805-1
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发表时间：
2018-01-01
影响因子：
2.9
通讯作者：
Ma Feng
中科院分区：
文献类型：
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作者：
Li Miao;Li Ping;Ma Feng
为探讨马钱子碱对高转移性乳腺癌细胞株MDAMB-231和Hs578-T迁移、侵袭、黏附及上皮向间充质转化(EMT)标志物和基质金属蛋白酶(MMPs)表达的影响，将高转移性乳腺癌细胞株MDAMB-231和Hs578-T分为4组：对照组(0.1%二甲基亚砜)、25、50和100mU/L组。细胞存活率用CellTiter-GloA(R)发光细胞存活率测定。采用划痕愈合实验和Tanswell迁移实验检测不同浓度马钱子碱对细胞迁移能力的影响。采用集落形成实验、跨孔侵袭实验和黏附实验分别检测细胞的增殖率、侵袭能力和黏附能力。用实时定量逆转录聚合酶链式反应和Western blotting检测EMT标志物MMP2和MMP9的蛋白和mRNA的表达。与对照组相比，马钱子碱对细胞的存活和增殖影响不大(P>；0.05)，但使MDA-MB-231和Hs578-T细胞的迁移、侵袭和黏附能力呈剂量依赖性下降(P<；0.01)。此外，马钱子碱还上调了MDA-MB-231和Hs578-T细胞中E-钙粘附素和β-连环素等EMT标志物的蛋白和mRNA水平，降低了间充质标志物波形蛋白和纤维连接蛋白的蛋白和mRNA水平，以及基质金属蛋白酶-2和基质金属蛋白酶-9的表达(P均<0.01)。
To examine the effect of brucine on the migration, invasion, adhesion and expressions of epithelial-to-mesenchymal transition (EMT) markers and matrix metalloproteinases (MMPs) in the highly metastatic breast cancer cell lines MDA-MB-231 and Hs578-T.MDA-MB-231 and Hs578-T cells were divided to 4 groups: the control group (0.1% DMSO), and 25, 50 and 100 mu mol/L brucine groups. The cell viability was determined using a CellTiter-GloA (R) luminescent cell viability. The scratch wound healing assay and tanswell migration assay were used to determine the migration ability of these cells treated by different concentrations of brucine. The proliferation rate, invasive potential and adhesive ability were respectively performed by colony formation assay, transwell invasion assay and adhension assay. The protein and mRNA expressions of EMT biomarkers, MMP-2 and MMP-9 were investigated by real-time reverse transcription polymerase chain reaction and Western blot.Compared with the control group, brucine had little effect on cell viability or proliferation (P > 0.05), but led to a dose-dependent decrease on migration, invasion, adhension of MDA-MB-231 and Hs578-T cells (P < 0.01). Furthermore, brucine increased the protein and mRNA levels of EMT markers such as E-cadherin and beta-catenin in MDA-MB-231 and Hs578-T cells, and decreased the protein and mRNA levels of mesenychmal markers such as vimentin and fibronectin, as well as the expressions of MMP-2 and MMP-9 (all P < 0.01).Brucine inhibited triple negative breast cancer cells metastasis potentially through EMT reversion and MMP-2 and MMP-9 inhibition.