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The Inhibitory effect of Antisense Oligodeoxynucleotides on PLOD2 Gene and Type ⅠCollagen Expression

Author: TanBin
Tutor: LuoZhongJin；ZhangJiXiang
School: Jiangxi Medical College
Course: Internal scientific digest
Keywords: Oligodeoxynucleotide Antisense The lysine hydroxylase 2 gene Collagen type I Liver fibrosis
CLC: R346
Type: Master's thesis
Year: 2005
Downloads: 36
Quote: 0
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Abstract


Antisense oligodeoxynucleotide (oligodeoxynucleotide, ODN) Objective To observe the different sequences of mouse fibroblasts ( NIH/3T3 ) PLOD2 ( procollage -lysine - oxoglutarate5 - dioxygenase ) mRNA and Ι collagen expression of the inhibitory effect . Method NIH/3T3 fibroblasts seeded in 6- well plate , by adding three different antisense ODN sequence ( for the translation initiation portion , for the first and second external connecting portion of exon for translation termination unit ) and a public The missense chain . After 5% CO2 37 ℃ incubation , the cells were collected for extraction of total RNA and protein , and then detected by RT-PCR method PLOD2mRNA type Ι collagen α1 chain (COLIα1) mRNA expression of β-actin expression as an internal control ; another Ι collagen expression detected by Western blot . The statistical analysis of variance between the results row group . 1 impact PLOD2mRNA, COLIα1mRNA : connecting portion of the translation initiation portion of the first exon and second exon , antisense ODN after the action of the translation termination portion PLOD2 inhibition rates were 0.57 ± 0.05,0.22 ± 0.05,0.31 ± 0.06. Antisense ODN group PLOD2mRNA expression was significantly inhibited ( P lt; 0.05 ) . Among them, the strongest inhibition of the translation initiation antisense ODN performance . Antisense and missense ODN the group COLIα1/β-actin standard values ??were 1.08 ± 0.05,1.08 ± 0.04,1.10 ± 0.02,1.13 ± 0.01, the control group was 1.14 ± 0.04 , antisense group compared with the control group, no significant decline , no significant difference ( P > 0.05 ) between the groups . Ι collagen expression : the translation initiation portion , the first exon and second outer exon connecting portion , termination codon portion after antisense ODN the Ι - type collagen inhibition rates were 0.44 ± 0.01,0.18 ± 0.04,0.23 ± 0.03. Antisense ODN could significantly inhibit the expression of the Ι type collagen (P lt; 0.05). Which inhibition strongest the antisense ODN performance translation initiation site . Conclusions antisense ODN significantly inhibited NIH/3T3 cells in mice PLOD2mRNA and Ι collagen expression . Antisense ODN sequence of PLOD2mRNA and Ι collagen expression

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