The Inhibitory Effect Of Combined P53 And AS Transfecting In HL60 Cell

ObjectiveTo observe the inhibitory effect and the expression levels of VEGF,bax,bcl-2 of combined p53 and AS transfecting in HL60 cell. To discuss the possibility on treatment in acute leukemia of combined genes.MethodsTransfect the p53 and AS into HL60 cell with lipofectamine. After transfection, RT-PCR was used to verify whether the target gene had been transfected and expressed or not.Cell clones trial, MTT growth curve and flow cytometry assay were used analyze to the inhibitory effect of combined p53 and AS transfecting in HL60 cell. RT-PCR and immunohistochemistry staining were used to analyze the expression levels of VEGF,bax,bcl-2 of combined p53 and AS transfecting in HL60 cell. The date were analysised by spss10.0.Results1. RT-PCR show: p53 and AS had been transfected and expressed in HL60 cell.2. Cell clones trial show: After transfection, the number of all cell clones were reduce(p<0.05), compared with the empty gene transfect group p<0.05, compared with the single gene transfect group p<0.05(n=3), compared with was evidence(p<0.05) and the cell clones became small.3. MTT growth curve show: After transfection, compared with the empty gene transfect group p<0.05, compared with the single gene transfect group p<0.05, compared with the empty gene transfect group, the cells in the single gene(p53 or AS)transfect group and the two genes(p53 and AS) transfect group grew slowly and longer(p<0.05).They were suppressedby the two genes and inhibition of the combined genes was more powerful than a single one(p<0.05).4. Flow cytometry assay show: In the single gene(p53 or AS)transfect group and the two genes(p53 and AS) transfect group could apper differ peak value of apoptosis, compared with the single gene transfect group, it was more evidence in the two genes(p53 and AS) transfect group, the inhibition effect of cooperate with combined p53 and AS transfecting in HL60 cell.5. Immunohistochemistry staining show: After transfection, the expression levels of VEGF and bcl-2 were lower, the expression levels of bax was higher than before transfection.ClonculsionsThe inhibition effect of cooperate with combined p53 and AS transfecting in HL60 cell. After transfection, the expression levels of VEGF and bcl-2 were lower, the expression levels of bax was higher. It may be one of the mechanism by the effect of bcl-2/bax rate.