Effect of baicalin on proliferation and apoptosis of HeLa cells

OBJECTIVE To investigate the effect of baicalin on proliferation and apoptosis of HeLa cells and its mechanism.METHODS CCK-8assay was performed to detect the proliferation of HeLa cells after cells were treated with different concentration of baicalin(5,10,20,40,80,160μg/mL)for 24 h,48hand 72 h.After treatment with 40μg/mL baicalin for 24 h,the morphological changes of HeLa cells were examined by light microscopy and nucleolus morphological changes were observed under fluorescence microscope after Hoechst 33342 staining.The cell early apoptosis was tested by Annexin Ⅴ-FITC/PI doublestaining analysis.Western blot assay was carried out to examine the protein expression of Caspase-3,Caspase-8and Caspase-9.RESULTS Baicalin obviously inhibited the proliferation of HeLa cells in a doseand time-dependent manner(P0.05).When HeLa cells were treated with 40μg/mL baicalin for 24 h,part of HeLa cells became small and round,even formed apoptotic bodies,exhibiting typical morphological features of apoptosis under light micrograph.Under inverted fluorescence microscope after Hoechst 33342 staining,the nuclei of HeLa cells became condensed or fragmented.When HeLa cells were treated with 40μg/mL of baicalin for 24 hand 48h,the ratio of early apoptotic cells reached(10.5±1.5)%,(21.8±2.4)%,respectively vs(1.3±0.9)%in control,the differences had statistical significance(P0.05).Western blot results showed that the cleaved Caspase-3,Caspase-8and Caspase-9were observed to be upregulated more evidently after treatment of HeLa cells with baicalin.The expression of them raised from0.09,0.04 and 0.12,reapsetively to 0.65,1.22,1.10 after treatment for 48 h.CONCLUSION Our finndings showed that baicalin can significantly inhibit the proliferation of HeLa cells and induced the apoptosis of HeLa cells,which may occur through activation of capsases.