Research Of Proliferation And Apoptosis Effects Of Andrographolide On Human Prostate Cancer DU145Cells

ObjectiveTo investigate the effect of Andro on proliferation and apoptosis of humanprostate cancer DU145cells in vitro，and make a preliminary discussion on itspossible mechanism.Methods(1)To observe the cell morphologic change induced by Andro(5.0μmol/L) for24h under invert microscope；(2)Using MTT method to messure DU145cellsgrowth inhibition rate after the cells were intervened with Andro in differentconcentration groups for24，48and72h，and calculate the value of IC50；(3)Propidium iodide(PI) single-staining flow cytometry assay method was used toanalyze cell cycles of DU145cells intervened with different Andro concentrationgroups for24h(；4)Flow cytometry was used to detect apoptosis of the cells withAnnexin V-FITC/PI double staining；(5)the expression of NF-κB、Bcl-2andCaspase-3protein were determined by western blot.Result(1)After being treated with Andro(5μmol/L) for24h，the cells showed typicalapoptotic morphological changes compare with untreated group：more cells lostconnection，lost normal spindle morphology，bacame rounder and smaller，andthe count decreased significantly；(2)MTT assay showed that Andro inhibitedDU145cells’ proliferation in time-and concentration-dependent manner. The inhibiting concentration of50%cell growth(IC50) at24、48and72h was(51.41±4.13)、(18.74±2.29) and (9.73±2.35) μmol/L；(3)The results detectedby Flow cytometry instrument showed that cell percentage of G0/G1phase inDU145cells were increased and cell percentage of S phase was reduced，withstatistical significance (P<0.05)；(4) Being treated with three differentconcentration of Andro for24h， the rate of apoptosis in each group were(8.39±0.76)%，(31.37±1.75)%and (36.41±1.31)%，and there were significantdifferences compared with control group (1.87±0.21)%，all P<0.05；(5)Westernblot showed that Andro could reduce the expression of NF-κB、Bcl-2protein，and increase the expression of Caspase-3protein in a concentration-dependentmanner，and there were significant differences comparing with the controlgroup(P<0.05).ConclusionAndrographolide could block cell cycle in G0/G1phase and inhibit theproliferation of DU145cells in vitro； Andro could induce the apoptosis ofDU145cells， its mechanism may be related with down-regulation of theexpression of NF-κB、Bcl-2and up-regulation of the expression of Caspase-3protein.