Effect of Cynanoside H on proliferation and metastasis of triple negative breast cancer MDA-MB-231 cells and its mechanism
Objective To investigate the effect of Cynanoside H on the proliferation and metastasis of triple negative breast cancer(TNBC)MDA-MB-231 cells and its mechanism,so as to provide an experimental basis for the development of TNBC therapeutics.Methods MDA-MB-231 cells were randomly divided into one control group(without Cynanoside H)and three test groups including 2.5,5 and 10 μmol/L Cynanoside H dose groups.The effect of Cynanoside H on the proliferation of MDA-MB-231 cells was detected by MTT assay and clone formation assay,while the effect on the cell cycle was detected by flow cytometry,and the effect on the metastasis of MDA-MB-231 cells was measured by wound healing assay,Transwell migration and invasion assay.In addition,the effects of Cynanoside H on the expression of cell cycle(c-Myc,CDK1,CDK2 and cyclin E1),metastasis(E-cadherin,Vimentin and β-catenin)and PDGFRB/JAK2/STAT3 pathway related proteins(PDGFRB,p-JAK2,JAK2,p-STAT3 and STAT3)in MDA-MB-231 cells were determined by Western blot.Results The levels of cell viability(t=4.598,19.77 and 53.43,respectively,each P＜0.05),growth(36 h:t=8.256,11.57 and 12.16,respectively,each P＜0.05;48 h:t=10.49,22.49 and 19.63,respectively,each P＜0.01;72 h:t=50.20,28.84 and 15.83,respectively,each P＜0.01;96 h:t=11.18,18.35 and 20.92,respectively,each P＜0.01)and clone forma-tion(t=4.618,9.821 and 12.14,respectively,each P＜0.05)of 2.5,5 and 10 μmol/L dose groups were significantly lower than those of the control group.Compared with the control group,the proportion of cells in S phase of three test group increased in a concentration-dependent manner(48 h:t=6.316,8.156 and 14.11,respectively,each P＜0.05;72 h:t=7.231,15.36 and 25.16,respectively,each P＜0.05),and the expression of c-Myc(5 and 10 μmol/L:t=10.39 and 12.18,P＜0.05 and＜0.01,respectively),CDK1(t=3.777,5.069 and 6.974,respectively,each P＜0.05),CDK2(5 and 10 μmol/L:t=12.72 and 19.43,respectively,each P＜0.01)and cyclinE1(t=3.813 and 15.23,respectively,each P＜0.01)was down-regulated at the protein level.Compared with the control group,each test group significantly inhibited wound healing(48 h:t=6.969,56.16 and 27.73,respectively,each P＜0.05;72 h:t=8.619,22.12 and 32.15,respec-tively,each P＜0.05),migration(t=9.817,14.74 and 19.39,respectively,each P＜0.01)and invasion(t=5.614,13.85 and 14.22,respectively,each P＜0.01)of MDA-MB-231 cells,and up-regulated the expression of E-cadherin(10 μmol/L:t=11.79,P＜0.01),down-regulated the expression of Vimentin(5 and 10 μmol/L:t=12.05 and 13.02,respectively,each P＜0.01)and(3-catenin(5 and 10 μmol/L:t=4.516 and 9.305,respectively,each P＜0.01)at the protein level.In addition,the test groups of Cynanoside H could significantly down-regulate the protein expression of PDGFRB(5 and 10 μmol/L:t=7.083 and 18.87,respectively,each P＜0.01)and inhibit the protein levels of p-JAK2(t=4.050,10.95 and 11.05,respectively,each P＜0.05)and p-STAT3(5 and 10 μmol/L:t=15.25 and 25.89,respectively,each P＜0.01),but had no effect on the total protein levels of JAK2 and STAT3.Conclusion Cynanoside H inhibits the proliferation of breast cancer MDA-MB-231 cells by inducing S-phase cell cycle arrest and reverses the epithelial-mesenchymal transition(EMT)to inhibit cell metastasis.These effects may be mediated by down-regulating PDGFRB/JAK2/STAT3 signaling pathway.
Cynanoside HTriple negative breast cancer(TNBC)MDA-MB-231 cellsCell proliferationCell metastasis
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