Effect of Skp2 antisense oligodeoxynucleotide on growth and proliferation of colorectal cancer SW 480 cells / 重庆医学

ABSTRACT

Objective To investigate the effect of S-phase kinase-associated protein 2 antisense oligodeoxynucleotide (Skp2 ASODN) on the growth and proliferation of colorectal cancer cells SW480 and its mechanism .Methods The liposome-mediated dif-ferent concentrations of Skp2 ASODN was adopted to transfect SW480 cells ,the final concentrations of 0 .050 ,0 .125 ,0 .250 ,0 .500 , 1 .000 ,2 .000 ,4 .000 μmol/L were respectively set up with Skp2 nonsense oligodeoxynucleotides(NSODN) and blank group as con-trol .Then the inhibited effect of growth and proliferation of colorectal cancer cells were measured by the inverted microscope obser-vationandmethylthiazolyltetrazolium(MTT),thecellcyclewassurveyedbytheflowcytometry(FCM).TheexpressionofmRNA and protein of Skp2 and P27kip1 were inspected by reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochem-istry methods .Results The inverted microscope observed that the SW480 cells had no change in form and grew at a slower speed . When the Skp2 ASODN concentration reached 0 .125 μmol/L ,the inhibition rate was 14 .48% (P0 .05) ,but its protein expression was elevates obviously (P<0 .01) .Conclusion Skp2 ASODN can inhibit the growth and proliferation of SW480 cells .Its possible mechanism is that the ubiq-uitin degradation of the Skp2 to P27kip1 is decreased by the gene occlusion effect of Skp2 ASODN ,thus the cell cycle progression is arrested .