Anti-breast Cancer Activities in Vitro of Oxymatrine Combined with Bevacizumab： An Exploration Focusing on EMT Regulation Through Wnt/β-catenin / 中国实验方剂学杂志

ABSTRACT

Objective:To observe the effect of oxymatrine （OM） combined with bevacizumab （ BV ） on the proliferation， invasion， and migration of breast cancer MCF-7 cells and explore the mechanism of OM in regulating BV-induced epithelial-mesenchymal transition （EMT） based on the Wnt/<italic>β</italic>-catenin signaling pathway. Method:The effect of different concentrations of OM（0， 0.5， 1.0， 2.0， 4.0， 8.0， 16.0 mmol·L^-1）and BV（0， 0.25×10^-4， 0.50×10^-4， 1.00×10^-4， 2.00×10^-4， 4.00×10^-4， and 8.00×10^-4 mmol·L^-1）on the proliferation of MCF-7 cells were detected by cell counting kit-8（CCK-8）assay. The effect of OM（4.0 mmol·L^-1） combined with BV（2.00×10^-4 mmol·L^-1）on the invasion and migration of MCF-7 cells were observed in transwell and scratch repair tests. Western blot was conducted to investigate the effect of OM（4.0 mmol·L^-1）combined with BV （2.00×10^-4 mmol·L^-1） on proliferation-related proteins in MCF-7 cells， followed by the detection of the expression levels of Wnt/<italic>β</italic>-catenin signaling pathway- and EMT-related proteins. Result:Compared with the blank group， OM （2.0，4.0，8.0，16.0 mmol·L^-1） inhibited the proliferation of MCF-7 cells in a concentration-dependent manner （<italic>P</italic><0.01）， while BV did not show the inhibitory effect against the proliferation of MCF-7 cells. The inhibitory effect of the combination of the two drugs on the proliferation of MCF-7 cells was not significantly different from that of OM. Compared with the blank group， OM significantly reduced the migration distance of MCF-7 cells and the number of invaded cells（<italic>P</italic><0.01）， while BV increased the migration distance of MCF-7 cells and the number of invaded cells （<italic>P</italic><0.05，<italic>P</italic><0.01）. Compared with BV， its combination with OM significantly inhibited the invasion and migration of MCF-7 cells induced by BV （<italic>P</italic><0.01）. Compared with the blank group， both OM and the combined medication obviously inhibited the phosphorylation of proliferation-related protein kinase B（Akt） and extracellular-signal-regulated protein kinase 1/2 （ERK1/2）in MCF-7 cells （<italic>P</italic><0.01） and down-regulated the protein expression levels of <italic>β</italic>-catenin， proto-oncogene （c-Myc）， CD44， and G₁/S-specific cyclin D₁ in Wnt/<italic>β</italic>-catenin signaling pathway （<italic>P</italic><0.05，<italic>P</italic><0.01）. Besides， OM and the combination of two drugs both significantly reduced the protein expression levels of calcium-dependent cell adhesion protein <italic>N</italic>-cadherin and Vimentin in EMT， whereas increased the expression of calcium-dependent cell adhesion protein E-cadherin（<italic>P</italic><0.01）. However， the expression of the above-mentioned proteins in the BV group was reversed （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:After the combination with BV， OM plays an anti-breast cancer role by effectively inhibiting the activation of Wnt/<italic>β</italic>-catenin pathway induced by BV and reversing EMT.