Li Miao, Li Ping, Zhang Mei, Ma Feng
Department of Geriatrics, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui Province, 233000, China.
Department of Chinese Medicine Tumor, the Affiliated Provincial Hospital of Anhui Medical University, Hefei, 230001, China.
Chin J Integr Med. 2018 Jan;24(1):40-46. doi: 10.1007/s11655-017-2805-1. Epub 2017 Aug 9.
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 μmol/L brucine groups. The cell viability was determined using a CellTiter-Glo® 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 β-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.
研究马钱子碱对高转移性乳腺癌细胞系MDA-MB-231和Hs578-T迁移、侵袭、黏附以及上皮-间质转化（EMT）标志物和基质金属蛋白酶（MMPs）表达的影响。
将MDA-MB-231和Hs578-T细胞分为4组：对照组（0.1%二甲基亚砜）以及25、50和100μmol/L马钱子碱组。使用CellTiter-Glo®发光细胞活力检测法测定细胞活力。采用划痕伤口愈合试验和Transwell迁移试验检测不同浓度马钱子碱处理后这些细胞的迁移能力。分别通过集落形成试验、Transwell侵袭试验和黏附试验检测增殖率、侵袭潜能和黏附能力。通过实时逆转录聚合酶链反应和蛋白质印迹法研究EMT生物标志物、MMP-2和MMP-9的蛋白质和mRNA表达。
与对照组相比，马钱子碱对细胞活力或增殖影响较小（P>0.05），但导致MDA-MB-231和Hs578-T细胞的迁移、侵袭和黏附呈剂量依赖性降低（P<0.01）。此外，马钱子碱使MDA-MB-231和Hs578-T细胞中E-钙黏蛋白和β-连环蛋白等EMT标志物的蛋白质和mRNA水平升高，使波形蛋白和纤连蛋白等间充质标志物的蛋白质和mRNA水平以及MMP-2和MMP-9的表达降低（均P<0.01）。
马钱子碱可能通过EMT逆转以及抑制MMP-2和MMP-9来抑制三阴性乳腺癌细胞转移。
