A Study On Transfection Of VEGF And/or Survivin SiRNA In A549Cells And Its Biological Responses

Objective：To investigate the transfection and its biological responses of VEGFand/or survivin siRNA in A549cells and if co-transfection of VEGF and survivinsiRNAgets better biological responses.Methods: NSCLC A549cells were cultured and the siRNA targeting VEGF and/orsurvivin was transfected with liposome entrapment. Five groups were employed:transfection with VEGF siRNA, survivin siRNA, VEGF siRNA and survivin siRNA,negative control siRNA(nc) and liposome only(blank control, bc) respectively. Thedosage of siRNA in each group was50nmol/L. The mRNA expression of VEGF andsurvivin was detected by RT-qPCR and their protein expression was detected byWestern blot. The inhibition of cell proliferation was examined by CCK-8and theapoptosis change was detected by flow cytometry.Results：1. Based on RT-qPCR and western blot, compared to negative and blankcontrols, the expressions of VEGF mRNA and its protein were significantlydown-regulated in48hours after transfection with VEGF siRNA or VEGF siRNA andsurvivin siRNA, of which the former group was much lower down. The expressions ofsurvivin mRNA and its protein were also significantly inhibited in48hours aftertransfection with survivin siRNA or VEGF siRNA and survivin siRNA, of which thetwo groups have no significant difference.2. CCK-8assay showed that compared tonegative and blank controls, the proliferation of A549cells was significantly inhibitedafter transfected with VEGF siRNA, survivin siRNA, VEGF siRNA and survivinsiRNA, with much more significance in the latter two groups.3. Flow Cytometry assayshowed that compared to negative and blank controls, the apoptotic rates were increasedin groups of transfection with survivin siRNA or VEGF siRNA and survivin siRNA, ofwhich the former group was more significant.Conclusion：1.VEGF siRNA or VEGF siRNA and survivin siRNA transfection was capable of down-regulating the expressions of VEGF mRNA and its protein, withmuch more significance in the former group. Survivin siRNA or VEGF siRNA andsurvivin siRNA transfection can also down-regulate the expressions of survivin mRNAand its protein.2.Transfection with VEGF siRNA or survivin siRNA or VEGF siRNAand survivin siRNA was able to inhibit the proliferation of cells, with much moresignificance in the latter two groups, however there was no significant differencebetween them.3.Survivin siRNA or VEGF siRNA and survivin siRNA trasfection canenhance the apoptosis of cells, with more significance in the the former group.4.Whilethe co-gene transfection of VEGF and survivin failed to show better biological efficacycompared to single gene transfection, it is still possible to show some positive responseif we improve the sequence of siRNAand further our study in other cell lines.