The Effect And Mechanism Of BF12 On The Proliferation And Apoptosis Of Cervical Cancer And Breast Cancer Cells

Objective:To study the mechanism of the benzo[b]furan CA-4 derivative?BF12[?E?-3-?6-methoxy-2-?1-?3,4,5-trimethoxyphenyl?vinyl?benzofuran-5-yl?carboxylic acid]?on proliferation and apoptosis in cervical cells?SiHa/HeLa?and breast cancer cells?MCF-7/MDA-MB-231?.Methods:?1?MTT assay screened the activity of CA-4derivatives against cancer cells?SiHa/HeLa/MCF-7/MDA-MB-231?.?2?PI single staining detected cell cycle distribution;western blot and immunofluorescence assays observed microtubule polymerization effect.?3?Tubule forming test estimated the tubulization ability of HUVECs,and the level of VEGF in breast tumor cell culture medium was detected by ELISA.?4?Hoechst 33342 nucleic acid staining observed the morphological changes,and Annexin V-FITC/PI double staining detected the apoptosis-inducing effect.?5?the molecular docking between BF12 and PI3K??,?,?,??proteins was carried out by the Autodock vina software,and the structure-activity relationship of antitumor activity was analyzed.western blot assay detected the expression level of PI3K/Akt/mTOR pathway proteins and apoptosis-related proteins.Scratch test observed breast cancer cells migration.?6?The effect of BF12 on autophagy was detected by immunofluorescence,electron microscope and western blot tests in breast cancer cells.Results:?1?The IC₅₀ of BF12 to SiHa/HeLa/MCF-7/MDA-MB-231 were 1.10/1.06/0.89/4.77?M,respectively.?2?BF12 selectively blocked the mitosis G2/M phase.Abnormal spindle formation in MCF-7cells and destructive microtubule cytoskeleton in SiHa/HeLa/MDA-MB-231 cells were observated by the Laser confocal.The level of polymerized?-/?-tubulin was decreasing?p<0.05?.?3?BF12 could effectively inhibit the formation of tubules and reduce the level of VEGF secretion?p<0.05?in MCF-7 and MDA-MB-231 cells.?4?Hoechst staining showed typical characteristics of apoptosis,which was induced in a concentration-dependent manner.?5?The molecular docking results showed that the four subunits of PI3K and BF12 could stably bind,and the binding free energy was-8.0,-7.7,-7.8,-6.9 kcal/mol,respectively.BF12 could down-regulate the level of proteins p-PI3K,p-Akt,p-mTOR,p-P70S6K,bcl-2,and up-regulate bax,p53,caspase-3/-9.The compound could also inhibit the migration of MCF-7 and MDA-MB-231 cells.?6?Autophagy fluorescence aggregates,autophagosomes and autophagy lysosomes were observated,and up-regulated expression level of LC3 and beclin-1 could be seen in breast cancer cells.Conclusion:BF12 synergized anti-cervical cancer and anti-breast cancer cells proliferation by inhibiting cell microtubule polymerization and the PI3K/Akt/mTOR signaling pathway;BF12 could also stimulate the autophagy and inhibit the migration by inhibiting the PI3K/Akt/mTOR signaling pathway,further revealing the mechanism of promoting apoptosis in breast cancer cells.