Effect Of RNAi-mediated Silencing Of Transforming Growth Factor-beta Receptor1 Gene On EMT Of Human Colon Cancer Lovo Cells

ObjectiveTo determine the effect of siliencing transforming growth factor-beta receptor1?TGFBR1?gene on proliferation and apoptosis and EMT of colon cancer Lovo cells in vitro.To investigate the mechanism of TGFBR1 in the development of colon c ancer cell's invasion and metastasis.MestodsThe recombinant siRNA TGFBR1-silencer and TGFBR1 vector targeting the T-GFBR1 gene was constructed.Human colon cancer Lovo cells were transfected with TGFBR1-silencer or TGFBR1-vector as the silencer and vector groups.The untransf-ected Lovo cells was as thenormal groups.At the same time,the transfection efficien-cy of siRNAin Lovo cells was identified by setting up the Cy3 fluorescence contr-olgroup.In the light of the three groups,TGFBR1,Twist,Snail,E-cadherin?CDH1?,Vime ntin?VIM?'s mRNA abundance and protein expression level were assessed by realti-meRT-PCR and western-blot.Cell proliferation was by CCK-8 assay.Cell cycle proge-ss and apoptosis were by flow cytometric assay in untransfected and transfected L-ovo cells.ResultsCompared with control vector-transfected and untransfectedLovo cells,Lovo cells transfected with TGFBR1-silencer had significantly lower TGFBR1 mRNA abund-an ce?0.2132±0.012,0.5987±0.031,0.6065±0.026,P<0.05??Twist?0.2352±0.022,0.5372±0.023,0.5813±0.017,P<0.05??Snail?0.2549±0.015,0.6135±0.014,0.5491±0.011,P<0.05??CD H1?0.5421±0.021,0.2125±0.013,0.2343±0.015,P<0.05??VIM?0.2054±0.012,0.5532±0.016,0.5117±0.019,P<0.05?;and a significantly lower proliferation activity,24h?0.431±0.037,0.213±0.057,0.387±0.107,P<0.05?,48h?0.783±0.102,0.486±0.158,0.805±0.112,P<0.05?;Its protein expression lever was also dcreased,TGFBR1?0.2085±0.016,0.5254±0.023,0.4937±0.036,P<0.05??Twist?0.1569±0.015,0.4891±0.027,0.5237±0.041,P<0.05??Snail?0.2016±0.021,0.5217±0.016,0.5814±0.026,P<0.05??VIM?0.2537±0.031,0.7952±0.027,0.7136±0.019,P<0.05?;except CDH1?0.3571±0.018,0.1772±0.052,0.1654±0.038,P<0.05?.TGFBR1 gene resulted significant increases in the percentage of cells i-n G₁/G₀phase,?S:?23.48±3.97?%vs?38.27±4.67?%,P<0.01?,and in apoptosisis alsoincrea sed,as compared with control vector-transfected and untransfected Lovo cells.ConclusionRNAi-mediated silencing of the TGFBR1gene results in human colon cancer c-ell proliferation,Garrest,EMT and apoptosis,thereby possessing a therapeutic potential.