Inhibitory Effect Of Silymarin On Human Osteosarcoma Saos-2 Cells And Its Mechanism

Objective:To study effect of silymarin on proliferation and apoptosis of human osteosarcoma saos-2 cells and further explore the role of gene P21?Bcl-2?Bax and Caspase-3 in it.Methods:(1)Human osteosarcoma saos-2 cells were cultured at logarithmic phase,the stduy was divided into control group and different concentration groups of drug,the morphologic changes of cells were observed by inverted phase contrast microscope.(2)Proliferation of cells were tested by CCK-8 assay.(3)Apoptosis of cells were detected by Annexin V-FITC/PI assay.(4)Expression level of anti-proliferation gene P21,and apoptosis related gene Caspase-3,Bax,Bcl-2mRN A were measured by RT-PCR.(5)Expression level of P21,Caspase-3 protein were measured by Western Blot.Results:(1)Control group cells grew well,d ifferent concentration groups of drug of cells becomed small and round,presented apoptosis-like change,the higher the drug concentration,the more obvious the morphological changes of cells.(2)CCK-8 assay test showed that proliferation of cells of drug groups was inhibited,presented time-and dose-dependent manners.(3)Annexin V-FITC/PI assay test showed that the number of apoptotic cells of drug groups was increased obviously,presented dose-dependent manner.(4)The results of RT-PCR showed that compared with comtrol group,expression level of anti-proliferation gene P21 and pro-apoptosis gene Caspase-3,Bax of drug groups were increased gradually(P<0.05),while anti-apoptosis gene Bcl-2 was decreased gradually(P<0.05).(5)The results of Western Blot showed that compared with comtrol group,expression level of P21,Caspase-3 protein were increased gradually(P<0.05).Conclusion:(1)Silymarin can inhibit human osteosarcoma saos-2 cells proliferation and induce its apoptosis.(2)Silymarin inhibit human osteosarcoma saos-2 cells by up-regulating express level of gene P21?Caspase-3?Bax and down-regulating express level of gene Bcl-2.