The Inhibitory Effect Of Antisense Oligodeoxynucleotides On PLOD2 Gene And Type â… Collagen Expression

objective To observe the inhibitory effects of antisenseoligonucleotides(ODN) on expression of PLOD2(procollage-lysine 2-oxoglutarate5-dioxygenase 2) and type Ιcollagen mRNA and protein by NIH/3T3 fibroblasts.Methods NIH/3T3 fibroblasts were cultured with DMEM mediumcontaining different antisense ODN(To translation initiation, To the joint exon1 andexon2, To translation terminal) and missense ODN. The total RNAs and proteinfrom the cells were extracted. Then, the PLOD2 and type Ιcollagen mRNA wereexamined with RT-PCR. The amount of type I collagen protein was measured bywestern blot.Results The Antisense ODN had obviously inhibitory effect on PLOD2gene and type I collagen protein expression(P<0.05), with the ratio of 0.57±0.05,0.22±0.05,0.31±0.06 mRNA level, 0.44±0.01,0.18±0.04,0.23±0.03 on protein level, respectively. Antisense ODN to translation initiation showedthe strongest inhibitory effect, compared with other two groups. then, type Icollagen mRNA levels(COLIα1/β-actin)were almost same(P﹥0.05), with ratioof 1.08±0.05,1.08±0.04,1.10±0.02,1.13±0.01,1.14±0.04.Conclusion Antisense ODN would effectively inhibit the expressions ofPLOD2 mRNA and type I collagen protein , with the strongest inhibitory effect byantisense ODN at translate initiation. Then,The Antisense ODN had not almostinhibitory effect on type I collagen gene.