A Study On The Effect Of TIC10 Against PC3 Human Prostate Cancer Cells In Vitro And In Vivo

Objective To observe the effect of TIC10 intervention on the vitality,migration and invasion of human prostate cancer cell line PC3 cultured in vitro and in vivo and the possible involvement mechanisms of apoptosis.The antitumor activity of TIC10 in vivo is studied by establishing a subcutaneous xenograft tumor model in nude mice.To investigate the potential anti-prostate cancer activity by TIC10 and provide a new experimental evidence for the clinical treatment of prostate cancer.Methods The viability,migration and invasion abilities of PC3 human prostate cancer cells were detected by MTS assay,scratch assay and Transwell assay,respectively.The apoptosis of PC3 cells were detected by Annexin V-FITC apoptosis detection assay.The mRNA levels of caspase-3,caspase-8,caspase-9,Bax,Bok,and Bcl-2 in cells were measured by quantitative real-time PCR(qRTPCR).Western Blot analysis was utilized to examine TRAIL,Akt,phosphorylated Akt(p-Akt),ERK1/2 and p-ERK1/2 expression levels.And in vivo,the effect of TIC10 on tumor growth was examined by using xenograft model of PC3 cells in nude mice.Upon tumor formation,mice were administered either PBS(1x)or TIC10(10 mg/kg)[intraperitoneally(i.p.)] once every 2 days for 18 days.Results When the concentration of TIC10 was less than 0.625?M,there was no significant difference in the cell viability between concentration and time effect(P>0.05).If the concentration of TIC10 was more than 0.625?M,it significantly inhibited human prostate cancer cell line PC3's viability after 96 h,and the inhibition gradually increased with the drug concentration(P<0.01).Exposure to TIC10 inhibited the mRNA expression levels of Bcl-2 and increased the expression of caspase-3,caspase-8,caspase-9,Bax and Bok to promote cell apoptosis in a dose-dependent manner(P<0.05).Moreover,on the molecular level,treatment with TIC10 leaded to a posttranslational decrease of p-Akt and pERK1/2 and up-regulation of TRAIL in a dose-dependent manner(P<0.05).However,TIC10 had no significant influence on the expression of Akt and ERK1/2 proteins.In vivo experiments,we observed that the volume of xenograft in the treated group was significantly smaller than that in the control group(P<0.01).Conclusions1.TIC10 could inhibit the viability,migration and invasion of prostate cancer PC3 cells.2.TIC10 induced apoptosis of PC3 cells via both extrinsic(caspase-8-dependent)and intrinsic(caspase-9-dependent)apoptosis pathways.The potential mechanism is to inactive Akt and ERK signaling,and up-regulate TRAIL expression.3.TIC10 markedly inhibited PC3 xenograft growth in nude mice.