Effects of PolyI: C on Mice Prostate Cancer Archive - Cancer ...

[Abstract]

PolyI:C is an artificial mimic of viral RNA composed withpolyinosinic?polycytidylic acid. It is well known that polyI:C not only caninduce both human and animal cell lines to secrete and synthese interferon butalso can regulate the cellular activites of immune system.So it has beenconsidered as an appropriate interferon inducer and an immune antitumordrugs .In this study, we attempt to observe the effects of polyI:C on mouseprostate cancer RM-1cell line in vivo and in vitro , then elucidate themechanism of it. Part 1 Methods: MTT assay was used to measure the cell proliferation and inhibitionrates;Fluorescence-activated cell sorting (FACS) and DNA ladder strap wereused to analyze the cell cycle distribution profile and apoptosis;RT-PCR wasused to evaluate the influence of PolyI:C on mRNA expression ofBcl-2,Bax,Capase-3 .Results:(1)Effect of polyI:polyC on RM-1cell viability :Cellular viability resultsshowed that, in the presence of different concentrations of polyI:C(100ng/ml,200ng/ml, 400ng/ml) , the MTT reduction was markedly decreased .Thepercentage of cell inhibition was approximately(11.05?0.04)%,(13.1?0.03)% and ( 17.84?0.04 ) % at 24h.The percentage of cell inhibition wasapproximately(23.86?0.09)%,(34.32?0.08)% and ( 52.31?0.06)% at 48hand(47.32?0.02)%,60.33?0.02)% and(71.48?0.01)% at 72 h respectivelyAtthe same time, RM-1 cell cultures became less confluent as some cells roundedand detached from the culture plates, compared with the control .(2)Effects of PolyI:C on Apoptosis: The specific DNA ladder straps wasobserved in all treated groups through 1.5% agarose gel electrophoresis aftercultured with polyI:C for 48h and the apoptosis was in dose-time-depandentway.while there are no specific DNA ladder straps in the control.Flow Cytometryshows PolyI:C induced apoptosis and caused cells significantly accumulated in G1phase compared with the control.( 3 ) Effects of PolyI:C on Bcl-2,Bax and Capase-3 inRM-1cells :RT-PCR showed that, compared with the control, the express ofBcl-2 and the ration of Bcl-2/Bax were significantly decreased ,whereas theexpress of Bax and Capase-3 were significantly increased in PolyI:C group.Discussion:Apoptosis is a basic biological process that plays an important role in cellturnover in normal tissues, and may also activate cell suicide in tumor tissues8.Inmany cases, PCD involves a characteristic set of mortphological events, knownas apoptosis, which include plasma membrane blebbing, cell shrinkage,chromatin condensation, and nuclear fragmentation, followed ultimately by thebudding off of cellular fragments9.In this study, we observe polyI:C can directlyinhibit RM-1 cell growth and cause cycle-specific apoptosis .RT-PCR showedtthat the express of Bax and Capase-3were increased in the presence of PolyI:Cbut the express of Bcl-2 and the ratio of Bcl-2/bax were decrease Thus a majorantitumor mechanism of polyI:C involved in seems to be associate with itsmodulation of the Bcl-2/Bax pathway, interference with caspases.Part2&3Method:Established prostate carcinoma models by inoculating tumor cells tosubcutaneous tissue of mice;according to tumor weight randomly divided miceinto two groups;Measured the tumor relative volumn and mice weight tocalculate the inhibition rates of polyi:c;Drew the tumor growth curves and micesurvival curves;H-E staining and immunohistochemical SP were used toevaluate the change of bcl-2 , bax ,caspase-3 ,VEGF and eNOS protein ofprostate carcinoma tissue;TdT-mediated X-dU TP nick end labeling(TUNEL)technique was used to stain the apoptosis cells in situ.Result:(1) Effects of PolyI:C on antiproliferation:The body weight of polyI:Cgroup and the control is 27.68?3.49g and 28.64?3.02g separately. This resultshow that there is no significant difference between two groups( P >0.05).Butthe tumor weight of polyI:C group and the control is 1.58?0.67 and 4.14?1.56separately and the tumor index of of polyI:C group and the controlis14.45%?1.06%? 5.42%?0.17% separately.There is significant difference intumor weight and the tumor index between two groups( P

Title: Effects of PolyI: C on Mice Prostate Cancer

Category: Breast Cancer

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