Inhibitions Of Salinomycin On Prostate Cancer PC-3Cells And Microspheres In Vitro

Objective: To observe the impact of salinomycin on the proliferation of humanprostate cancer PC-3cells, and exploring the salinomycin influence on the CD44+and CD133+cells double positive in PC-3cells and the formation of microspheres,Find out genome that closely related to the role of salinomycin from PC-3cells.Materials and methods:1: Adopting MTT method to test the activity of PC-3cells in different concentrations of alinomycin, measuring the absorbance value under570nm wavelength by the Microplate reader, drawing the curve and calculat the IC50.2:The cultured PC-3cells are divided into three groups, Group A for blankcomparing, group B for9uM DMSO, group C for9uM salinomycin, each groupare given anti-human CD133mouse monoclonal antibody and CD44mouseanti-human monoclonal antibody and two antibodies at the same time, detected theproportion of positive cells in each group before and after the impact of salinomycinby flow cytometry. Analysis the data by Statistical software.3: Tumor microsphereswas cultured from cancer stem cell,and observed the formation and depolymerizationof tumor microsphere under the microscope.4:Detect gene expression profiles ofPC-3cell before and after the salinomycin by microarray, difference analysis gene byTreeview software, the above tune or down more than10times as the standard toscreen the target genome which closely related to salinomycin, statistical analysis tofilter out the most representative of the two target genes and explore gene function. Result(1) Various concentrations of salinomycin inhibit the proliferation of the PC-3cell,the IC50are9.12μmol/ml.(2) Compared with the control group, the proportion of CD44+, CD133+cells anddouble positive cells in salinomycin group were lower, the difference between themwas statistically significant (P <0.05).(3) Compared with the DMSO group, the proportion of CD44+, CD133+cells anddouble positive cells in salinomycin group were lower, the difference between themwas statistically significant (P <0.05).(4) Compared with the Blank control group, the proportion of CD44+, CD133+cellsand double positive cells in DMSO group were not significant decrease(P>0.05)(5)Salinomycin can promote tumor microspheres depolymerization, increase in sizeof the cells, swelling and death it.(6) Microarray test results: The number of15genes were down-regulated morethan10times, The number of34genes were up-regulated more than10times, andthe POSTN gene down13.2times, CALB2gene increased47.55times.Conclusion:(1)Salinomycin can effectively inhibit the proliferation of human prostate cancer PC-3cells in vitro.(2) Salinomycin significantly reduced the proportion of CD44+and CD133+cellsin PC-3cells, and promote tumor microspheres depolymerization, so, salinomycininhibit the growth of PC-3cells by kill tumor stem cells.(3)Salinomycin change the whole gene expression in PC-3cells，Significantlyup-regulated CALB2gene expression and the downward POSTN one.