Effect of p27mt gene on apoptosis of the colorectal cancer cell line Lovo

AIM:To construct p27mt recombinant adenovirus,transfect the colorectal cell line Lovo and observe the effects of p27mt on Lovo cell apoptosis and cell cycle inhibition.METHODS:We constructed recombinant adenovirus containing p27mt by homologous recombination in bacteria.The colorectal cancer cell line Lovo was infected with recombinant replication-defective adenovirus Ad-p27mt,and expression of p27mt was determined by Western blotting;the inhibitory effect of p27mt on Lovo cells was detected by cytometry.Cell cycle was determined by flow cytometry.DNA fragment analysis identif ied the occurrence of apoptosis.RESULTS:The recombinant adenovirus which already contained p27mt target gene was successfully constructed.When multiplicity of infection was ≥ 50,the infection efficiency was 100%.After transfection of Lovo cells with Ad-p27mt the cells had high p27 expression which was identified by immunoblotting assay.PI staining and ? ow cytometry showed that 77.96% of colorectal cancer cells were inhibited in phase G0/G1,while in the Ad-LacZ group and blank control group,27.57% and 25.29% cells were inhibited in the same phase,respectively.DNA fragment analysis,flow cytometry and TUNEL assay demonstrated that p27mt is able to induce apoptosis in colorectal cancer cells.CONCLUSION:p27mt has an obvious blocking effect on colorectal cancer cell cycle,and most cells were inhibited in phase G0/G1.Therefore,p27mt can induce apoptosis in colorectal cells.