Effects Of Shrna Targeting TPX2on The Apoptotic Rate And Sensitivity To Paclitaxel Of A549Cells

Objective: To study the effect of short hairpin RNA (shRNA) targetedTPX2on apoptosis and sensitivity to paclitaxel of Lung AdenocarcinamaA549cells.Method: shRNA-TPX2expression vector constructed and confirmedby sequencing, was transfected into A549cells using lipofectamine2000and G418was selected and cloned. The expression levels of TPX2mRNAand protein were detected by RT-PCR and Western blotting, cell cycle andapoptotic rate were analyzed by Flow cytometry, sensitivity to paclitaxel inA549were determined by MTT assay. The levels of Aurora-A, P53, Bcl-2,LRP and MRP mRNA were detected by RT-PCR.Results: After shRNA-TPX2expression vector transfection, RT-PCRand Western blotting assays showed that there was a significant decrease inTPX2mRNA and protein levels. Flow cytometry showed that the cells majorstagnative stayed in the phase S and the apoptotic rate increased obviously.The difference was significant compared with control group (P<0.05).ShRNA-TPX2significantly up-regulated the expression of P53mRNA and down-regulated the expressions of Aurora-A and Bcl-2mRNAs comparedwith control group (P<0.05) but had no effect on LRP and MPR mRNAsConclusion: shRNA-TPX2expression vector can significantly suppressTPX2expression, block cells in S phase, induce apoptosis and enhances thesensitivity to paclitaxel of A549cells. This effect may be related withup-regulation of the expression of P53and down-regulation of Bcl-2.