Effect Of Salidroside On Human Breast Cancer MDA-MB-435 Cells And Preliminary Analysis Of The Differential Expression Proteins

Objective: To explore the effects and mechanism of Salidroside against human breast cancer, cell proliferation, apoptosis, cell migration and invasion were detected in human breast cancer MDA-MB-435 cells treated with Salidroside(Sal). The proteomics technologies were used to investigate the differential expression proteins intervented by Sal in MDA-MB-435 cells, for further studying the molecular mechanism of Sal involved in the treatment of human breast cancer.Methods: The human breast cancer MDA-MB-435 cells were cultured in vitro. MDA-MB-435 cells were treated with the different concentrations of Salidroside(0.01mg/ml, 0.1mg/ml, 1mg/ml, 10mg/ml, 100mg/ml) for 24 h, and the half maximal inhibitory concentration(IC50) of Salidroside on MDA-MB-435 cells was determined by Cell count kit(CCK8). MDA-MB-435 cells treated with IC50 of Sal were considered as the Sal group, which cells untreated with Sal were as the control group. Moreover, cell proliferation was detected in MDA-MB-435 cells treated with the half maximal inhibitory concentration(IC50) of Salidroside at 24 h, 48 h and 72 h time points by CCK8 assay, and the cell shape was observed using light microscope. Flow cytometry technology was used to detect apoptosis and cell cycle. And Transwell assay was used to observe the cell migration and invasion in MDA-MB-435 cells. Besides, the proteomics technologies were used to establish 2-DE profiling on MDA-MB-435 cells treated with 4mg/ml Sal, analysis and identified six different expression proteins.Results: The IC50 of Sal on human breast cancer MDA-MB-435 cells was 4mg/ml. Using CCK8 assay, it was shown that the MDA-MB-435 cells proliferation treated with the IC50 of Sal were depressed at the time point of 24 h, 48 h and 72 h with the comparison to control group(P<0.05). And the inhibition rate in Sal group was higher than that of the control group in the time dependent manner. Flow cytometry was used to analyze cells apoptosis by AnnexinⅤ-FITC / PI apoptosis assay kit, the results demonstrated that the percentage of early apoptotic cells was increased to(3.31±0.21)% in Sal group, compared with(1.26±0.13)% in control group(P<0.05), while the late apoptosis and necrosis of MDA-MB-435 cells was induced to(28.80±2.15)% in Sal group, compared with(5.56±0.78)% in control group(P<0.01). And the results of FITC proved that the rate of G0/G1 phase was enhanced and the rate of S and G2/M phase were decreased comparison with that of the control group. Cell migration and invasion assay in vitro revealed that the average number of invaded and migrated MDA-MB-435 cells in Sal group cells of outer surface in migrant assay:(39.60±3.37); cells in invasive assay:(4.20±1.55) significantly decreased in comparison with the control group cells of outer surface in migrant assay:(142.20±12.99)(P<0.01); cells in invasive assay:(13.00±3.09)(P<0.05). Moreover, the 2-DE profiling of MDA-MB-435 cells treated with Sal was established using the proteomics technologies. Then the six differential expression proteins were identified by mass spectrometry and bioinformatics analysis, which related with the process of components of the cell membrane, the regulation of ion channel, cell signal transduction, cell apoptosis and motility.Conclusion: 1.Salidroside treatment in MDA-MB-435 cells promoted apoptosis, and inhibited cell proliferation, arrested cell cycle and repressed the motility and invasion significantly. 2. 2-DE profilings in MDA-MB-435 cells treated with Salidroside were established, and six differential expression proteins were identified.