Effect Of Co-transfection Of PTEN Gene On The Bladder Cancer T24Cells

ObjectiveTo explore the relationship between the expression of gene PTEN andapoptosis of bladder cancer cells by studying the expression of tumorsuppressor gene PTEN in bladder cancer cells,and to observe the inhibitioneffect bladder cancer of gene PTEN on T24cells.MethodsRecombinant plasmid was transfected into bladder cancer T24cells whichwere lack of gene PTEN.There are three groups in this experiment: group A(containing pEGFP-PTEN plasmid) group B (containing pEGFP-N1emptyplasmid) and group C (containing the same content of liposomes).The inhibitioneffect of PTEN gene on bladder cancer T24cells was observed Theexpression of target gene was determine by RT-PCR method.A method ofWestern blotting method was used to determine the concentration of PTENprotein and and expression of Phospho-Akt protein.The inhibition effect on thecells’ growth was determined by the establishment of cells colony.The effect ofPTEN gene on the apoptosis of T24cells was determined by the double dyeflow cytometryResultsThe successful expression of PTEN gene in T24cells was observed by thefluorescence microscope.The stripes of three groups was observed respectivelyat the length about1200bp accroding to the consequence ofRT-PCR.Compared to group B and group C， the OD value of group A wassignificantly increased(P<0.05）.The expression of PTEN protein in group A was significantly increased compared with this in group B and group C(P<0.05）. Theexpression of Phospho-Akt protein was significantly reduced in group Acompared with this in group B and group C(P<0.05）.The scale of cells colonywas significantly reduced in group A compared with this in group B and groupC(P<0.05）.Apoptosis rate of cells in group A was significantly increasedcompared with this in group B and group C(P<0.05）ConclusionTarget gene was transfected into bladder cancer T24cells by recombinantplasmid and expressed stablely and the proliferation of cancer cells wasinhibited by this stable expression.Westernblot tests shows that in the aspect ofcontrol of apoptosis may be wild-type PTEN gene lowered Phospholated Aktexpression level.By looking at the mechanism of apoptosis, tthe apoptosis ofbladder cancer cells may be related to the inhibition effect of gene PTEN onthe PI3K/AKT pathway.This experiment provied the evidence for the genetherap and it lay the foundation of clinical application of gene therapy.