Abstract
Objective To construct the plasmid of B lymphocytes/leukemia-2 (bcl-2) and observe its expression in U251 cells.Methods Plasmid of bcl-2 was constructed and transfected into U251 cells by FUGENE HD.G418 was used to kill the cells that were transfected unsucessfully.Western blotting and reverse transcriptase polymerase chain reaction (RT-PCR) were used to observe the expression of bcl-2.Three groups were set up:control group,N1 group and bcl-2 group.Methyl thiazol tetrazolium (MTT) assay was used to measure the viability of U251 cells in N1 group and bcl-2 group.Results The plasmid PEGFPbcl-2 and bcl-2-U251 cells were constructed successfully.The expression of bcl-2 in control group,N1 group and bcl-2 group was 0.012 ±0.003,0.011 ±0.003 and 0.115 ±0.025,and the viability of U251 cells was 1.015 ±0.005,1.120 ± 0.004 and 3.312 ± 0.004,respectively.There was significant difference between bcl-2 group and control group (P ＜ 0.05).Conclusion The plasmid PEGFP-bcl-2 was constructed successfully and the overexpression system is also established,and bcl-2 overexpression can increase the viability of U251 cells. Key words: B lymphocytes/leukemia-2 ; Glioma; Plasmid
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